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Journal Cover Journal of Cellular Physiology
  [SJR: 1.842]   [H-I: 139]   [7 followers]  Follow
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   ISSN (Print) 0021-9541 - ISSN (Online) 1097-4652
   Published by John Wiley and Sons Homepage  [1589 journals]
  • Effects of mTOR/NF-κB signaling pathway and high thoracic epidural
           anesthesia on myocardial ischemia-reperfusion injury via autophagy in rats
    • Authors: Wei-Qiang Huang; Jian-lin Wen, Ri-Qi Lin, Peng Wei, Feng Huang
      Abstract: We investigated the role of mammalian target of rapamycin/nuclear factor-kappa B (mTOR/NF-κB) signaling pathway in high thoracic epidural anesthesia (HTEA) against myocardial ischemia-reperfusion (I/R) injury in rats. The rat model of myocardial I/R injury was established. Ninety rats were divided into the normal, sham, I/R, eHTEA, the PDTC and HTEA + PDTC groups. ELISA was applied to detect cardiac function indexes. HE staining was conducted to observe histopathological changes of myocardial tissues, and TTC staining was performed to detect the myocardial infarction size. TUNEL staining was adopted to detect the cell apoptosis rate. The mRNA and protein levels of mTOR, NF-κB, Fasl, Bcl-2 and Bax, and LC3-I, LC3-II, BNIP3 and Atg5 were detected by RT-qPCR and western blotting, respectively. The findings indicated that compared with the normal and sham groups, the I/R, PDTC and HTEA groups showed the larger myocardial infarction size and increased cell apoptosis rate, while the results in the HTEA + PDTC group were opposite. Compared with the normal and sham groups, the I/R group showed reduced mRNA and protein levels of Bcl-2, LC3, BNIP3, and Atg5, and elevated mRNA and protein levels of mTOR, p50, p65, Bax and Fasl, while the HTEA + PDTC group revealed the opposite results, and the PDTC and HTEA group revealed the increased mRNA and protein levels of Bcl-2, LC3, BNIP3, Atg5, mTOR, p50, p65, Bax and Fasl. These results prove that the inhibition of mTOR/NF-κB signaling pathway potentiates HTEA against myocardial IR injury by autophagy and apoptosis in rats. This article is protected by copyright. All rights reserved
      PubDate: 2017-12-05T09:25:36.855686-05:
      DOI: 10.1002/jcp.26320
  • Network Analysis of Hippocampal Neurons by Microelectrode Array in the
           Presence of HIV-1 Tat and Cocaine
    • Authors: Taha Mohseni Ahooyi; Masoud Shekarabi, Emilie A. Decoppet, T. Dianne Langford, Kamel Khalili
      Abstract: HIV-associated neurocognitive disorders affecting greater than 30% of patients are caused by HIV-1 infection of the CNS, and in part, include neurotoxic effects of the viral transactivator of transcription, Tat protein. In addition to increasing the risk for becoming HIV infected, cocaine abuse enhances the neuropathogenic impacts of HIV-1. To investigate the outcome of Tat and cocaine interference in the hippocampal neuronal network, microelectrode arrays were employed to develop a systematic framework to assess the rank-cross-correlation coefficient in cultured hippocampal neurons. Tat and cocaine differentially disturbed neuronal spiking rates, amplitude, synchronous activity and oscillations within the hippocampal neuronal network via potentiation of inhibitory neurotransmission. The Tat-mediated impairment of neuronal spiking was reversible by removal of Tat, which restored neuronal activity. The presence of astrocytes co-cultured with neuronal networks diminished the effects of Tat and cocaine on neuron function suggesting a role for astrocytes in stabilizing neuronal behavior and increasing neuronal spontaneous activities such as bursting amplitude, frequency and wave propagation rate. Taken together, our studies indicate that the HIV protein Tat and cocaine impair hippocampal neuronal network functioning and that the presence of astrocytes alleviates network dysfunction pointing to a newly discovered pathway through which ionic homeostasis is maintained by neuron-glial crosstalk in the CNS. This article is protected by copyright. All rights reserved
      PubDate: 2017-12-05T09:25:25.942816-05:
      DOI: 10.1002/jcp.26322
  • MicroRNA-6869-5p acts as a tumor suppressor via targeting TLR4/NF-κB
           signaling pathway in colorectal cancer
    • Authors: Shushan Yan; Zengfang Wang, Quanhong Duan, Changcheng Wang, Donghua Xu
      Abstract: Many studies have implicated that microRNAs (miRNAs), as non-coding RNAs, play important roles in the development and progression of colorectal cancer (CRC). However, little is known about the role of a newly identified miRNA, miR-6869-5p, in CRC. We aim to investigate the modifying effects and underlying mechanisms of miR-6869-5 in colorectal carcinogenesis and progression. Significantly reduced levels of miR-6869-5p were observed in both serum exosomes tumor tissue samples from patients with CRC. The prediction of targets of miR-6869-5p in databases of targetscan, microRNA.ORG and miRDBA revealed that toll-like receptor 4 (TLR4) is a potential target for this miRNA. MiR-6869-5p could inhibit cell proliferation and the production of inflammatory cytokines (TNF-α and IL-6) in CRC cells via directly targeting TLR4. The protective effect of miR-6869-5p from colorectal carcinogenesis was dependent on TLR4/NF-κB signaling pathway. In addition, the 3-year survival was poor among CRC patients with decreased levels of miR-6869-5p in serum exosomes. Thus, miR-6869-5p may serve as a tumor suppressor in CRC, and serum exosomal miR-6869-5p is a promising circulating biomarker for the prediction of CRC prognosis. This article is protected by copyright. All rights reserved
      PubDate: 2017-12-05T08:40:59.869063-05:
      DOI: 10.1002/jcp.26316
  • Hormone regulates endometrial function via cooperation of endoplasmic
           reticulum stress and mTOR-autophagy
    • Authors: Diqi Yang; Tingting Jiang, Jianguo Liu, Jin Hong, Pengfei Lin, Huatao Chen, Dong Zhou, Keqiong Tang, Aihua Wang, Yaping Jin
      Abstract: In ruminant, the receptive endometrium and the elongation of the hatched blastocyst are required to complete the process of implantation. However, the mechanisms regulating goat endometrial function during the peri-implantation period of pregnancy are still unclear. In this study, EECs were treated with progesterone, estradiol and interferon-tau (IFNT). We have found that endoplasmic reticulum (ER) stress were activated under hormones treatment. To identify the cellular mechanism of regulation of endometrial function, we investigated the effect of ER stress activator thapsigargin (TG) and inhibitor 4 phenyl butyric acid (4-PBA) on EECs. We found that TG, which activated the three branches of UPR, increased the expression of genes associated with promoting conceptus elongation and cellular attachment, significantly up-regualted the spheroid attachment rate and PGE2/PGF2α ratio. 4-PBA pre-treatment inhibited UPR and inhibited promoting conceptus elongation and cellular attachment related genes, but the spheroid attachment rate and PGE2/PGF2α ratio were not changed significantly. Moreover, knockdown of ATF6 via shATF6 promoted the conceptus elongation related genes, but increased the dissolution of the corpus luteum. Besides, blocking ATF6 attenuated autophagy by activating mammalian target of rapamycin (mTOR) pathway. Moreover, rapamycin (mTOR inhibitor) pre-treatment inhibited the expression of promoting conceptus elongation and increased PGE2/PGF2α ratio. Taken together, our study indicated that physiological level of ER stress may contribute to early pregnancy success, and ATF6 signaling pathway cooperated with autophagy to regulate endometrial function by modulating mTOR pathway. This article is protected by copyright. All rights reserved
      PubDate: 2017-12-05T08:40:57.056431-05:
      DOI: 10.1002/jcp.26315
  • MicroRNA-137 promotes endothelial progenitor cell proliferation and
           angiogenesis in cerebral ischemic stroke mice by targeting NR4A2 through
           the Notch pathway
    • Authors: Xing-Li Liu; Gang Wang, Wei Song, Wei-Xin Yang, Jian Hua, Liang Lyu
      Abstract: Cerebral ischemic stroke (CIS) is one of the common causes of death and disability worldwide. This study aims to investigate effect of miR-137 on endothelial progenitor cells and angiogenesis in CIS by targeting NR4A2 via the Notch pathway. Brain tissues were extracted from CIS and normal mice. Immunohistochemistry was used to determine positive rate of NR4A2 expression. Serum VEGF, Ang, HGF and IκBα levels were determined by ELISA. RT-qPCR and western blotting were used to determine expression of related factors. Endothelial progenitor cells in CIS mice were treated and grouped into blank, NC, miR-137 mimics, miR-137 inhibitors, siRNA-NR4A2 and miR-137 inhibitors + siRNA-NR4A2 groups, and cells in normal mice into normal group. Proliferation and apoptosis were determined by MTT and flow cytometry, respectively. NR4A2 protein expression was strongly positive in CIS mice, which showed higher serum levels of VEGF, Ang and HGF but lower IκBα than normal mice. Compared with normal group, the rest groups (endothelial progenitor cells from CIS mice) showed decreased expressions of miR-137, Hes1, Hes5 and IκBα but elevated NR4A2, Notch, Jagged1, Hey-2, VEGF, Ang and HGF, inhibited proliferation and enhanced apoptosis. Compared with blank and NC groups, the miR-137 mimics and siRNA-NR4A2 groups exhibited increased expression of miR-137, Hes1, Hes5 and IκBα, but decreased NR4A2, Notch, Jagged1 and Hey-2, with enhanced proliferation and attenuated apoptosis. The miR-137 inhibitors group reversed the conditions. miR-137 enhances the endothelial progenitor cell proliferation and angiogenesis in CIS mice by targeting NR4A2 through the Notch signaling pathway. This article is protected by copyright. All rights reserved
      PubDate: 2017-12-05T08:40:24.488494-05:
      DOI: 10.1002/jcp.26312
  • Dickkopf Homolog 3 (DKK3): a candidate for detection and treatment of
    • Authors: Leila Hamzehzadeh; Michele Caraglia, Stephen L. Atkin, Amirhossein Sahebkar
      Abstract: Wnt signaling is an evolutionary highly conserved pathway that is modulated by several inhibitors and activators, and plays a key role in numerous physiological processes. One of the extracellular Wnt inhibitors is the DKK (Dickkopf Homolog) family which has four members (Dkk1-4) and a unique Dkk3-related gene, Dkkl1 (soggy). DKK3 is a divergent member of the DKK protein family. Evidence suggests that DKK3 may serve as a potential therapeutic target in several types of human cancers. We review here the biological role of DKK3 as a tumor suppressor gene (TSG) or oncogene, and its correlation with various miRNAs. In addition, we discuss the role of polymorphisms and promoter methylation of the DKK3 gene, and of its expression in regulating cancer cell proliferation. Finally, we propose that DKK3 may be considered as both a biomarker and a therapeutic target in different cancers. This article is protected by copyright. All rights reserved
      PubDate: 2017-12-05T08:40:20.88251-05:0
      DOI: 10.1002/jcp.26313
  • Evaluation of RNA from human trabecular bone and identification of stable
           reference genes
    • Authors: Simona Cepollaro; Elena Della Bella, Dario Biase, Michela Visani, Milena Fini
      Abstract: The isolation of good quality RNA from tissues is an essential prerequisite for gene expression analysis to study pathophysiological processes. This study evaluated the RNA isolated from human trabecular bone and defined a set of stable reference genes. After pulverization, RNA was extracted with a phenol/chloroform method and then purified using silica columns. The A260/280 ratio, A260/230 ratio, RIN, and ribosomal ratio were measured to evaluate RNA quality and integrity. Moreover, the expression of six candidates was analyzed by qPCR and different algorithms were applied to assess reference gene stability. A good purity and quality of RNA was achieved according to A260/280 and A260/230 ratios, and RIN values. TBP, YWHAZ and PGK1 were the most stable reference genes that should be used for gene expression analysis. In summary, the method proposed is suitable for gene expression evaluation in human bone and a set of reliable reference genes has been identified. This article is protected by copyright. All rights reserved
      PubDate: 2017-12-05T08:35:31.954988-05:
      DOI: 10.1002/jcp.26319
  • Effects of poly(I:C) and MF59 co-adjuvants on immunogenicity and efficacy
           of survivin polypeptide immunogen against melanoma
    • Authors: Xiaoyu Jiang; Shanshan Guan, Yongbo Qiao, Xiao Li, Yan Xu, Lan Yang, Ziyu Kuai, Haihong Zhang, Yuhua Shi, Wei Kong, Yaming Shan, Hao Zhang
      Abstract: Malignant tumors pose a public health problem that jeopardizes human life and quality of living. At present, tumor vaccines in clinical research typically are aimed at stimulating the cellular immune response, while more effective vaccines should take into account the synergy between broad spectrum antibodies and high levels of cellular immunity. In this study, epitope peptides (68–81, 95–104, 80–88) of the tumor antigen survivin were chosen as immunogens and supplemented with poly(I:C) and/or MF59 adjuvant to evaluate the immune effects and anti-melanoma activities. The results indicated that poly(I:C) and MF59 could assist the survivin epitope peptide immunogen to control the tumor size, quality and volume in black melanoma mouse models. Analyses by antibody titering, antibody isotyping and ELISPOT suggested that the adjuvanted immunogen could induce humoral immunity in mice. Poly(I:C) and MF59 combined with survivin peptide 95–104 could effectively induce humoral immunity mediated by type 2 T helper (Th2) cells. This study provides a basis for candidate immunogen design based on survivin and provides support for tumor therapy that can induce a more balanced Th1/Th2 immune response. This article is protected by copyright. All rights reserved
      PubDate: 2017-12-05T08:35:21.392787-05:
      DOI: 10.1002/jcp.26317
  • A new parameter of growth inhibition for cell proliferation assays
    • Authors: Francesco P. Fiorentino; Luigi Bagella, Irene Marchesi
      Abstract: Cell proliferation assays are performed by four decades to test the anti-proliferative activity of natural products and synthetic compounds in cell cultures. In cancer research, they are widely employed to evaluate drug efficacy in in vitro tumor models, such as established cell lines, primary cultures, and recently developed three-dimensional tumor organoids. In this manuscript, we demonstrated that current employed parameters used by researchers to quantify in vitro growth inhibition, IC50 and GI50, lead to a misinterpretation of results based on the exponential, and not linear, proliferation of the cells in culture. Therefore, we introduce a new parameter for the analysis of growth inhibition in cell proliferation assays, termed relative population doubling capacity, that can be employed to properly quantify the anti-proliferative activity of tested compounds and to compare drug efficacy between distinct cell models.We introduce here a new parameter for the analysis of growth inhibition in cell proliferation assays, termed relative population doubling capacity, that can be used to properly quantify and compare the anti-proliferative activity of tested compounds on exponential growing cell models.
      PubDate: 2017-12-04T05:23:02.540617-05:
      DOI: 10.1002/jcp.26208
  • Differentially regulated gene expression in quiescence vs. senescence and
           identification of ARID5A as a quiescence associated marker
    • Authors: Tarique Anwar; Bijoya Sen, Savera Aggarwal, Rhisita Nath, Niteen Pathak, Ajay Katoch, Mohamed Aiyaz, Nirupma Trehanpati, Sanjeev Khosla, Gayatri Ramakrishna
      Abstract: In multicellular organisms majority of the cells remain in a non‐dividing states of either fully differentiated or quiescence (reversible) or senescence (irreversible) conditions. In the present study, gene expression signatures unique to quiescence and senescence were identified using microarray in osteosarcoma cell line, U2OS. Besides, it was also noted that certain genes and pathways like NOD pathway was shared by both the growth arrest conditions. A major highlight of the present study was increased expression of number of chemokines and cytokines in both quiescence and senescence. While senescence‐associated secretory phenotype (SASP) is well known, the quiescence‐associated secretory phenotype (QASP) is relatively unknown and appeared novel in this study. ARID5A, a subunit of SWI/SNF complex was identified as a quiescence associated gene. The endogenous expression of ARID5A increased during serum starved condition of quiescence. Overexpression of ARID5A resulted in more number of cells in G0/G1 phase of cell cycle. Further ARID5A overexpressing cells when subjected to serum starvation showed a pronounced secretory phenotype. Overall, the present work has identified gene expression signatures which can distinguish quiescence from senescence. This article is protected by copyright. All rights reserved
      PubDate: 2017-12-04T05:11:58.058834-05:
      DOI: 10.1002/jcp.26227
  • Annexin A1, Annexin A2, and Dyrk 1B are upregulated during GAS1-induced
           cell cycle arrest
    • Authors: Gilberto Pérez-Sánchez; Adriana Jiménez, Marco A. Quezada-Ramírez, Enrique Estudillo, Alberto E. Ayala-Sarmiento, Guillermo Mendoza-Hernández, Justino Hernández-Soto, Fidel C. Hernández-Hernández, Febe E. Cázares-Raga, Jose Segovia
      Abstract: GAS1 is a pleiotropic protein that has been investigated because of its ability to induce cell proliferation, cell arrest, and apoptosis, depending on the cellular or the physiological context in which it is expressed. At this point, we have information about the molecular mechanisms by which GAS1 induces proliferation and apoptosis; but very few studies have been focused on elucidating the mechanisms by which GAS1 induces cell arrest. With the aim of expanding our knowledge on this subject, we first focused our research on finding proteins that were preferentially expressed in cells arrested by serum deprivation. By using a proteomics approach and mass spectrometry analysis, we identified 17 proteins in the 2-DE protein profile of serum deprived NIH3T3 cells. Among them, Annexin A1 (Anxa1), Annexin A2 (Anxa2), dual specificity tyrosine-phosphorylation-regulated kinase 1B (Dyrk1B), and Eukaryotic translation initiation factor 3, F (eIf3f) were upregulated at transcriptional the level in proliferative NIH3T3 cells. Moreover, we demonstrated that Anxa1, Anxa2, and Dyrk1b are upregulated at both the transcriptional and translational levels by the overexpression of GAS1. Thus, our results suggest that the upregulation of Anxa1, Anxa2, and Dyrk1b could be related to the ability of GAS1 to induce cell arrest and maintain cell viability. Finally, we provided further evidence showing that GAS1 through Dyrk 1B leads not only to the arrest of NIH3T3 cells but also maintains cell viability.Two dimensional protein profiles in serum-deprived NIH3T3 cells revealed the upregulation of Gas1, Anxa1, Anxa 2, Dyrk 1B. Further experiments with GAS1-overexpressing cells demonstrated that GAS1 induces the upregulation of Anxa1, Anxa 2 and Dyrk 1B. Finally, we provide evidence showing that GAS1, through Dyrk 1B, leads not only NIH3T3 cells to cell arrest but also maintains cell viability.
      PubDate: 2017-12-04T05:10:54.438901-05:
      DOI: 10.1002/jcp.26226
  • Synergistic actions of Vitamin D and metformin on skeletal muscles and
           insulin resistance of Type 2 Diabetic Rats
    • Authors: Shaimaa N. Amin; Usama K. Hussein, Hanan D. Yassa, Sherif S. Hassan, Laila A. Rashed
      Abstract: Diabetes Mellitus is a chronic generalized disorder due to insulin insufficiency or resistance. Skeletal muscles represent one of the most important target organs that is affected by insulin signaling. The aim of the current work was to investigate the effect of metformin versus vitamin D (and also simultaneous administration) therapy in type 2 diabetic (T2D) rats on the state of the muscle and insulin sensitivity. Thirty six male rats constituted the animal model and have been divided into five groups: control, Diabetic, Diabetic+Metformin, Diabetic+ VitaminD, Diabetic+Metformin+Vitamin D. blood samples were taken for biochemical measurements of serum calcium, interleukin-6 (IL-6), Triglycerides (TG), glucose, insulin, and calculation of HOMA-IR, and then rats were sacrificed , dissected for removal of gastrocnemius muscle that is used for both biochemical, histopathological and electron microscopy examination. Oral administration of vitamin D alone or in combination with metformin improved insulin sensitivity in skeletal muscles, and sustained the metabolic complications along with muscle atrophy and inflammation in T2D rats. We demonstrated super-beneficial action on insulin resistance of additional vitamin D therapy in T2DM rats that were insufficiently controlled by metformin alone. This article is protected by copyright. All rights reserved
      PubDate: 2017-12-02T15:30:47.986625-05:
      DOI: 10.1002/jcp.26300
  • Using the Connectivity Map to discover compounds influencing human
           osteoblast differentiation
    • Authors: A M Brum; J Peppel, L Nguyen, A Aliev, M Schreuders-Koedam, T V Gajadien, C S Leije, A Kerkwijk, M Eijken, J P T M Leeuwen, B C J Eerden
      Abstract: Osteoporosis is a common skeletal disorder characterized by low bone mass leading to increased bone fragility and fracture susceptibility. Identification of factors influencing osteoblast differentiation and bone formation is very important. Previously, we identified parbendazole to be a novel compound that stimulates osteogenic differentiation of human mesenchymal stromal cells (hMSCs), using gene expression profiling and bioinformatic analyses, including the Connectivity Map (CMap), as an in‐silico approach. The aim for this paper is to identify additional compounds affecting osteoblast differentiation using the CMap. Gene expression profiling was performed on hMSCs differentiated to osteoblasts using Illumina microarrays. Our osteoblast gene signature, the top regulated genes 6‐hours after induction by dexamethasone, was uploaded into CMap ( Through this approach we identified compounds with gene signatures positively correlating (withaferin‐A, calcium folinate, amylocaine) or negatively correlating (salbutamol, metaraminol, diprophylline) to our osteoblast gene signature. All positively correlating compounds stimulated osteogenic differentiation, as indicated by increased mineralization compared to control treated cells. One of three negatively correlating compounds, salbutamol, inhibited dexamethasone‐induced osteoblastic differentiation, while the other two had no effect. Based on gene expression data of withaferin‐A and salbutamol, we identified HMOX1 and STC1 as being strongly differentially expressed . shRNA knockdown of HMOX1 or STC1 in hMSCs inhibited osteoblast differentiation. These results confirm that the CMap is a powerful approach to identify positively compounds that stimulate osteogenesis of hMSCs, and through this approach we can identify genes that play an important role in osteoblast differentiation and could be targets for novel bone anabolic therapies. This article is protected by copyright. All rights reserved
      PubDate: 2017-12-01T06:25:43.237584-05:
      DOI: 10.1002/jcp.26298
  • TGFβ signaling controls intrahepatic bile duct development may through
           regulating the Jagged1‐Notch‐Sox9 signaling axis
    • Authors: Wei Wang; Yuan Feng, Yasen Aimaiti, Xin Jin, Xixian Mao, Dewei Li
      Abstract: Due to the inherent limitations of the mouse models, the molecular mechanism of TGFβ signaling involved in the development of intrahepatic bile ducts (IHBDs) has been investigated little. Here, we investigated the role of TGFβ signaling and its regulatory mechanism in IHBDs development. We demonstrate that TGFβ signaling pathway activity is essential for IHBDs development. When blocking TGFβ signaling at E10.5, the number of bile ducts in hilum was reduced more than 2‐fold and number of CK19 positive chlangiocytes in periphery was reduced more than 3.5‐fold compared with controls. We also show that alpha‐smooth muscle actin (α‐SMA)‐immunoreactive cells are located in the portal vein mesenchyme (PVM) adjacent to the bile ducts during IHBDs development and identify the α‐SMA positive cells expressing the Notch ligand Jagged1 in the periportal area. Importantly, after blocking TGFβ signaling, the expression of Jagged1 was selectively decreased in the PVM but not in biliary epithelial cells (BECs), which is associated with the transformation of portal mesenchyme cells (PMCs) into portal myofibroblasts (PMFs). In addition, Sox9, which is downstream of Notch, is decreased after blocking the TGFβ signaling pathway in the liver. Our findings uncover a novel mechanism of TGFβ signaling in controlling the development of IHBDs may through regulating the Jagged1‐Notch‐Sox9 signaling axis. This article is protected by copyright. All rights reserved
      PubDate: 2017-12-01T00:25:51.662603-05:
      DOI: 10.1002/jcp.26304
  • Comparative analysis of single‐cell RNA sequencing data from mouse
           spermatogonial and mesenchymal stem cells to identify differentially
           expressed genes and transcriptional regulators of germline cells
    • Authors: Sajjad Sisakhtnezhad; Parvin Heshmati
      Abstract: Identifying effective internal factors for regulating germline commitment during development and for maintaining spermatogonial stem cells (SSCs) self‐renewal is important to understand the molecular basis of spermatogenesis process, and to develop new protocols for the production of the germline cells from other cell sources. Therefore, this study was designed to investigate single‐cell RNA‐sequencing data for identification of differentially expressed genes (DEGs) in 12 mouse‐derived single SSCs (mSSCs) in compare with 16 mouse‐derived single mesenchymal stem cells. We also aimed to find transcriptional regulators of DEGs. Collectively, 1584 up‐regulated DEGs were identified that are associated with 32 biological processes. Moreover, investigation of the expression profiles of genes including in spermatogenesis process revealed that Dazl, Ddx4, Sall4, Fkbp6, Tex15, Tex19.1, Rnf17, Piwil2, Taf7l, Zbtb16 and Cadm1 are presented in the first 30 up‐regulated DEGs. We also found 12 basal transcription factors (TFs) and 3 sequence‐specific TFs that control the expression of DEGs. Our findings also indicated that MEIS1, SMC3, TAF1, KAT2A, STAT3, GTF3C2, SIN3A, BDP1, PHC1 and EGR1 are the main central regulators of DEGs in mSSCs. In addition, we collectively detected 2 significant protein complexes in the protein‐protein interactions network for DEGs regulators. Finally, this study introduces the major upstream kinases for the main central regulators of DEGs and the components of core protein complexes. In conclusion, this study provides a molecular blueprint to uncover the molecular mechanisms behind the biology of SSCs and offers a list of candidate factors for cell type conversion approaches and production of germ cells. This article is protected by copyright. All rights reserved
      PubDate: 2017-12-01T00:25:48.534685-05:
      DOI: 10.1002/jcp.26303
  • Immunomodulatory effects of foreskin mesenchymal stromal cells
           (FSK‐MSCs) on natural killer (NK) cells
    • Authors: Mehdi Najar; Mohammad Fayyad-Kazan, Nathalie Meuleman, Dominique Bron, Hussein Fayyad-Kazan, Laurence Lagneaux
      Abstract: Foreskin‐mesenchymal stromal cells (FSK‐MSCs) are immune‐privileged thus making them valuable immunotherapeutic cell product. Characterization of the relationship between FSK‐MSCs and natural killer (NK) cells is essential to improve cell‐based therapy. In the present study, we studied for the first time FSK‐MSCs‐NK interaction and showed that the result of such cross talk was robustly dependent on the type of cytokines (IL‐2, IL‐12, IL‐15 and IL‐21) employed to activate NK cells. Distinctly activated‐NK cells showed uneven cytotoxicity against FSK‐MSCs, triggering their death in fine. The expression of different cell‐surface ligands (CD112, CD155, ULPB‐3) and receptors (LAIR, KIRs) ensuring such interaction was altered following co‐culture of both populations. Despite their partial negative effect on NK cell proliferation, FSK‐MSCs boosted the capacity of activated NK‐cells to secrete IFN‐γ and TNF‐α. Moreover, FSK‐MSCs enhanced degranulation of NK cells, reinforced secretion of perforin and granzymes, whilst only modestly increased ROS production. On the other hand, FSK‐MSCs‐mediated expression of C1 and B9 serpins was significantly lowered in the presence of activated NK cells. Altogether, our results highlight major immunological changes following FSK‐MSCs‐NK interaction. Understanding these outcomes will therefore enhance the value of the therapeutic strategy. This article is protected by copyright. All rights reserved
      PubDate: 2017-12-01T00:25:45.851765-05:
      DOI: 10.1002/jcp.26305
  • Narrower insight to SIRT1 role in cancer: A potential therapeutic target
           to control epithelial‐mesenchymal transition in cancer cells
    • Authors: Jalal Choupani; Sima Mansouri Derakhshan, Sahar Bayat, Mohammad Reza Alivand, Mahmood Shekari Khaniani
      Abstract: The epithelial–mesenchymal transition (EMT) is a highly networked cellular process which involves cell transition from the immotile epithelial to the motile mesenchymal phenotype, whereby cells lose their cell‐cell adhesion and cell polarity. This important process is one of the underlying mechanisms for enabling invasion and metastasis of cancer cells which is considered as malignant phase of tumor progression. However, the molecular mechanisms of this process are not fully clarified. It is reported that the nicotinamide adenine dinucleotide‐dependent histone deacetylase SIRT1 is associated with tumor metastasis through positive regulation of EMT in several types of cancers. Besides that, recent studies confirmed that up and down regulation of SIRT1 expression remarkably change the migration ability of different cancer cells in vitro and tumor metastasis in vivo. Also, according to this fact that carcinomas as the main human solid tumors, originate from different epithelial cell types, SIRT1 role in EMT has received a great attention due to its potential role in tumor development and metastasis. Therefore, SIRT1 has been proposed as a key regulator of cancer metastasis by promoting EMT, although little is known about the cleared effect of SIRT1 in this transition. Our aim in this review is to explain in more detail the role of SIRT1 in various signaling pathways related to carcinogenesis, with the focus on the promoting role of SIRT1 in EMT as a potential therapeutic target to control EMT and to prevent cancer progression. This article is protected by copyright. All rights reserved
      PubDate: 2017-12-01T00:22:31.895544-05:
      DOI: 10.1002/jcp.26302
  • Nanocurcumin restores aberrant miRNA expression profile in multiple
           sclerosis, randomized, double‐blind, placebo‐controlled trial
    • Authors: Sanam Dolati; Leili Aghebati Maleki, Majid Ahmadi, Faroogh Marofi, Zohreh Babaloo, Hormoz Ayramloo, Zahra Jafarisavari, Hamid Oskouei, Amir Afkham, Vahid Younesi, Mohammad Nouri, Mehdi Yousefi
      Abstract: Objective: In the current study, we aimed to identify nanocurcumin effects on microRNAs (miRNAs) in the peripheral blood of patients with relapsing‐remitting multiple sclerosis (RRMS). We intended to investigate the expression pattern of these miRNAs in experimental settings in vivo.Methods: The expression levels of the selected 27 miRNAs known to be involved in the regulation of immune responses were analyzed in 50 RRMS patients and 35 healthy controls. The miRNA expression profiles were investigated by quantitative PCR (qPCR) at baseline and after 6 months of nanocurcumin therapy.Results: Our data revealed that the expression of a number of microRNAs including miR‐16, miR‐17‐92, miR‐27, miR‐29b, miR‐126, miR‐128, miR‐132, miR‐155, miR‐326, miR‐550, miR‐15a, miR‐19b, miR‐106b, miR‐320a, miR‐363, miR‐31, miR‐181c, miR‐150, miR‐340, and miR‐599 is regulated by nanocurcumin.Interpretation: The results of the current work indicate that nanocurcumin is able to restore the expression pattern of dysregulated miRNAs in MS patients. We discovered that some miRNAs are deregulated in untreated patients compared with healthy controls and nanocurcumin‐treated patients. This is a new finding that might represent the potential contribution of these miRNAs to MS pathogenesis. Taken together, these data provide novel insights into miRNA‐dependent regulation of the function of B and T cells in MS disease and enrich our understanding of the effects mediated by a therapeutic approach that targets B and T cells. This article is protected by copyright. All rights reserved
      PubDate: 2017-12-01T00:22:29.44495-05:0
      DOI: 10.1002/jcp.26301
  • Effects of microRNA‐129 and its target gene c‐Fos on proliferation and
           apoptosis of hippocampal neurons in rats with epilepsy via the MAPK
           signaling pathway
    • Authors: Yu-Tong Zhang
      Abstract: This study aims to investigate the effect of microRNA‐129 (miR‐129) on proliferation and apoptosis of hippocampal neurons in epilepsy rats by targeting c‐Fos via the MAPK signaling pathway. Thirty rats were equally classified into a model group (successfully established as chronic epilepsy models) and a control group. miR‐129, c‐Fos, bax and MAPK expressions were detected by RT‐qPCR and Western blotting. Hippocampal neurons were assigned into normal, blank, negative control (NC), miR‐129 mimic, miR‐129 inhibitor, siRNA‐c‐Fos, miR‐129 inhibitor + siRNA‐c‐Fos groups. The targeting relationship between miR‐129 and c‐Fos was predicted and verified by bioinformatics websites and dual‐luciferase reporter gene assay. Cell proliferation after transfection was measured by MTT assay, and cell cycle and apoptosis by flow cytometry. c‐Fos is a potential target gene of miR‐129. Compared with the normal group, the other 6 groups showed a decreased miR‐129 expression, increased expressions of c‐Fos, Bax and MAPK, decreased proliferation, accelerated apoptosis, more cells in the G1 phase and fewer cells in the S phase. Compared with the blank and NC groups, the miR‐129 mimics group and the siRNA‐c‐Fos group showed increased miR‐129 expression, decreased c‐Fos, Bax and MAPK expressions, increased cells proliferation, and decreased cell apoptosis, fewer cells in the G1 phase and more cells in the S phase. However, the miR‐129 inhibitors groups showed reverse consequences. This study suggests that miR‐129 could inhibit the occurrence and development of epilepsy by repressing c‐Fos expression through inhibiting the MAPK signaling pathway. This article is protected by copyright. All rights reserved
      PubDate: 2017-11-30T08:16:19.028725-05:
      DOI: 10.1002/jcp.26297
  • Induced Mutation and Epigenetics Modification in Plants for crop
           improvement by targeting CRISPR/Cas9 technology
    • Authors: Muhammad Hafeez Ullah Khan; Shahid Ullah Khan, Ali Muhammad, Hu Limin, Yang Yang, Fan Chuchuan
      Abstract: Clustered regularly interspaced palindromic repeats associated protein Cas9 (CRISPR‐Cas9), originally an adaptive immunity system of prokaryotes, is revolutionizing genome editing technologies with minimal off‐targets in the present era. The CRISPR/CAS9 is now highly emergent, advanced and highly specific tool for genome engineering. The technology is widely used to animal and plant genomes to achieve desirable results. The present review will encompass how CRISPR‐Cas9 is revealing its beneficial role in characterizing plant genetic functions, genomic rearrangement, how it advances the site‐specific mutagenesis and epigenetics modification in plants to improve the yield of field crops with minimal side‐effects. The possible pitfalls of using and designing CRISPR‐Cas9 for plant genome editing are also discussed for its more appropriate applications in plant biology. Therefore, CRISPR/Cas9 system has multiple benefits that mostly scientists select for genome editing in several biological systems. This article is protected by copyright. All rights reserved
      PubDate: 2017-11-30T08:15:41.540565-05:
      DOI: 10.1002/jcp.26299
  • JAK3 and PI3K mediates bovine Interferon‐tau stimulated gene expression
           in the blood neutrophils
    • Authors: Aasif Ahmad Sheikh; O. K. Hooda, A. K. Dang
      Abstract: Interferon tau, a 23 KDa trophoblast derived protein diffuses out from the uterus into the circulation and leads to the expression of IFNτ stimulated genes viz. ISG15 and OAS1 in blood neutrophils. The IFNτ pathway is species as well as tissue specific. To unsnarl the IFNτ downstream signaling pathway, the blood neutrophils were incubated simultaneously with 10 ng/ml of Recombinant Bovine Interferon tau and the inhibitors of JAK2 (AG490), JAK3 (CP690550), p38 (SB202190), PI3K/Akt (LY294002) and MAPK/Erk (U0126) at specific doses for 4 hour duration. The IFNτ pathway was determined through real‐time gene expression of ISG15 and OAS1, immunocytochemistry of ISG15 and western blotting of ISG15, OAS1, pJAK3 and PI3K. The ISG15 and OAS1 expression decreased significantly (p
      PubDate: 2017-11-30T08:15:35.888702-05:
      DOI: 10.1002/jcp.26296
  • LncRNA‐PVT1 promotes pancreatic cancer cells proliferation and migration
           through acting as a molecular sponge to regulate miR‐448
    • Authors: Liang Zhao; Hongru Kong, Hongwei Sun, Zongjing Chen, Bicheng Chen, Mengtao Zhou
      Abstract: The identification and characterization of long non‐coding RNAs (lncRNAs) in diverse biological process has currently developed rapidly. LncRNA‐PVT1, located adjacent to the MYC locus on chromosomal region 8q24, has been reported to be associated with many biological processes. However, the function and mechanism of PVT1 in pancreatic carcinoma (PC) is poorly understood. In this present study, we first measured the level of PVT1 in the PC cell lines and tissues by quantitative real‐time PCR (qRT‐PCR), and then employed loss‐of‐function and gain‐of‐function approaches to explore the association between PVT1 expression levels and PC cell proliferation/migration ability. Furthermore, bioinformatics analysis was utilized to show that PVT1 contains binding site for miR‐448 and an inverse correlation between PVT1 and miR‐448 was obtained in PC specimens. Additionally, dual luciferase reporter assay, RNA‐binding protein immunoprecipitation (RIP) and applied biotin‐avidin pulldown system were applied to further confirm that PVT1 directly bind with microRNA binding site harboring in the PVT1 sequence. Then, SERBP1 was identified as a target of miR‐448 according to the gene expression array analysis of PC clinical samples. Together, we revealed that PVT1 functions as an endogenous “sponge” by competing for miR‐448 binding to regulate the miRNA target SERBP1 and, therefore, promotes the proliferation and migration of PC cells.lncRNA‐PVT1 functions as an endogenous “sponge” by competing for miR‐448 binding to regulate the miRNA target SERBP1 and, therefore, promotes the proliferation and migration of PC cells.
      PubDate: 2017-11-30T04:27:08.599791-05:
      DOI: 10.1002/jcp.26072
  • Exercise improves nicotine reward‐associated cognitive behaviors and
           related α7 nAChR‐mediated signal transduction in adolescent rats
    • Authors: Yuehui Zhou; Cuicui Li, Rena Li, Chenglin Zhou
      Abstract: The adolescent brain is vulnerable to long‐lasting cognitive disturbances resulting from nicotine exposure. Although exercise has been used as an intervention for ameliorating cognitive deficits in various disorders, the effect on cognitive changes induced by nicotine exposure and its underlying mechanisms remain unclear. The purpose of this study was to investigate the effects of exercise on nicotine reward‐associated cognitive behaviors in adolescent rats subjected to nicotine conditioned place preference paradigm (CPP). Male adolescent rats were trained on the Go/NoGo task, then subjected to nicotine CPP, and then randomly separated into four groups: sedentary (SED) and high‐ (HE), moderate‐ (ME) and low‐intensity (LE) exercise. Rats in exercise groups performed treadmill running 30 min daily for 10 days. Results showed that MEs had shorter escape latencies in the Morris water maze (MWM) test compared to SEDs. Although time spent and distance swam in the target quadrant significantly increased in both the MEs and HEs, the number of target quadrant crosses increased significantly only in MEs. MEs and HEs showed higher performance accuracy on NoGo and lower scores on CPP tasks. Expression of α7 nicotinic acetylcholine receptors (nAChRs) and downstream signaling molecules increased in MEs in prefrontal cortex but not hippocampus, with α7 nAChRs expression positively associated with NoGo accuracy and MWM probe test performance, but negatively correlated with CPP scores. The findings of this study suggest that moderate‐intensity exercise can improve nicotine induced cognitive behaviors, and implicates prefrontal cortical α7 nAChR‐mediated signal transduction as a possible mechanism. This article is protected by copyright. All rights reserved
      PubDate: 2017-11-29T09:12:31.031768-05:
      DOI: 10.1002/jcp.26295
  • Involvement of Dying Beta Cell Originated Messenger Molecules in
           Differentiation of Pancreatic Mesenchymal Stem Cells Under Glucotoxic and
           Glucolipotoxic Conditions
    • Authors: Selda Gezginci-Oktayoglu; Evren Onay-Ucar, Serap Sancar-Bas, Ayse Karatug-Kacar, Emine S. N. Arda, Sehnaz Bolkent
      Abstract: Beta cell mass regulation represents a critical issue for understanding and treatment of diabetes. The most important process in the development of diabetes is beta cell death, generally induced by glucotoxicity or glucolipotoxicity, and the regeneration mechanism of new beta cells that will replace dead beta cells is still not fully understood. The aim of this study was to investigate the generation mechanism of new beta cells by considering the compensation phase of type2 diabetes mellitus. Pancreatic islet derived mesenchymal stem cells (PI‐MSCs) were isolated from adult rats and characterized. Then, beta cells isolated from rats were co‐cultured with PI‐MSCs and they were exposed to glucotoxicity, lipotoxicity and glucolipotoxicity conditions for 72 hours. As the results apoptotic and necrotic cell death were increased in both PI‐MSCs and beta cells especially by the exposure of the co‐culture systems to glucotoxic and glucolipotoxic conditions. Glucotoxicity induced‐differentiated beta cells were functional due to their capability of insulin secretion in response to rising glucose concentrations. Moreover, beta cell proliferation was induced in the glucotoxicity‐treated co‐culture system whereas suppressed in lipotoxicity or glucolipotoxicity‐treated co‐culture systems. In addition, 11 novel proteins, that may release from dead beta cells and have the ability to stimulate PI‐MCs in the direction of differentiation, were determined in media of lipotoxicity or glucolipotoxicity‐treated co‐culture systems. In conclusion, these molecules were considered as important for understanding cellular mechanism of beta cell differentiation and diabetes. Thus, they may be potential targets for diagnosis and cellular or therapeutic treatment of diabetes. This article is protected by copyright. All rights reserved
      PubDate: 2017-11-28T04:16:40.518022-05:
      DOI: 10.1002/jcp.26242
  • Genetic and epigenetic factors influencing vitamin D status
    • Authors: Afsane Bahrami; Hamid Reza Sadeghnia, Seyed-Amir Tabatabaeizadeh, Hamidreza Bahrami-Taghanaki, Negin Behboodi, Habibollah Esmaeili, Gordon A. Ferns, Majid Ghayour Mobarhan, Amir Avan
      Abstract: The global prevalence of vitamin D deficiency appears to be increasing, and the impact of this on human health is important because of the association of vitamin D insufficiency with increased risk of osteoporosis, cardiovascular disease and some cancers. There are few studies on the genetic factors that can influence vitamin D levels. In particular, the data from twin and family‐based studies have reported that circulating vitamin D concentrations are partially determined by genetic factors. Moreover, it has been shown that genetic variants (e.g., mutation) and alteration (e.g., deletion, amplification, inversion) in genes involved in the metabolism, catabolism, transport, or binding of vitamin D to it receptor, might affect vitamin D level. However, the underlying genetic determinants of plasma 25‐hydroxyvitamin D3 [25(OH)D] concentrations remain to be elucidated. Furthermore, the association between epigenetic modifications such as DNA methylation and vitamin D level has now been reported in several studies. The aim of current review was to provide an overview of the possible value of loci associated to vitamin D metabolism, catabolism, and transport as well epigenetic modification and environmental factors influencing vitamin D status.
      PubDate: 2017-11-28T04:10:35.700377-05:
      DOI: 10.1002/jcp.26216
    • Authors: Michele Andreucci; Teresa Faga, Antonio Pisani, Raffaele Serra, Domenico Russo, Giovambattista De Sarro, Ashour Michael
      Abstract: Radiocontrast media (RCM)‐induced acute kidney injury (CI‐AKI) is a major clinical problem whose, pathophysiology is not well understood. Direct toxic effects on renal cells, possibly mediated by reactive oxygen species, have been postulated as contributing to CI‐AKI. We investigated the effect of quercetin on human renal proximal tubular (HK‐2) cells treated with the radiocontrast medium (RCM) sodium diatrizoate. Quercetin is the most widely studied flavonoid, and the most abundant flavonol present in foods. It has been suggested to have many health benefits, including angioprotective properties and anti‐cancer effects. These beneficial effects have been attributed to its antioxidant properties and its ability to modulate cell signaling pathways. Incubation of HK‐2 cells with 100μM quercetin caused a decrease in cell viability and pre‐treatment of HK‐2 cells with 100μM quercetin followed by incubation with 75mgI/ml sodium diatrizoate for 2h caused a decrease in cell viability which was worse than in cells treated with diatrizoate alone. However, further incubation of the cells (for 22 h) after removal of the diatrizoate and quercetin caused a recovery in cell viability in those cells previously treated with quercetin+diatrizoate and quercetin alone. Analysis of signaling molecules by Western blotting showed that in RCM‐treated cells receiving initial pre‐treatment with quercetin, followed by its removal, an increase in phosphorylation of Akt (Ser473), pSTAT3 (Tyr705) and FoxO3a (Thr32) as well as an induction of Pim‐1 and decrease in PARP1 cleavage were observed. Quercetin may alleviate the longer‐term toxic effects of RCM toxicity and its possible beneficial effects should be further investigated. This article is protected by copyright. All rights reserved
      PubDate: 2017-11-28T04:10:32.017784-05:
      DOI: 10.1002/jcp.26213
  • Cover Image, Volume 233, Number 3, March 2018
    • Authors: Meng-Hao Pan; Fei Wang, Yujie Lu, Feng Tang, Xing Duan, Yu Zhang, Bo Xiong, Shao-Chen Sun
      Abstract: Cover: The cover image, by Meng‐Hao Pan et al., is based on the Original Research Article, FHOD1 regulates cytoplasmic actin‐based spindle migration for mouse oocyte asymmetric cell division,
      DOI : 10.1002/jcp.26099.
      PubDate: 2017-11-26T08:49:21.118527-05:
  • C‐C motif chemokine ligand 23 abolishes ER stress‐ and LPS‐induced
           reduction in proliferation of bovine endometrial epithelial cells
    • Authors: Whasun Lim; Hyocheol Bae, Fuller W. Bazer, Gwonhwa Song
      Abstract: To reduce embryonic losses in domestic animals for economic production of livestock meat and milk, chemokines and their receptors are required for proper implantation and placentation during early pregnancy. In this study, we investigated the effects of C‐C‐motif chemokine ligand 23 (CCL23) on the proliferation of bovine endometrial (BEND) epithelial cells. CCL23 treatment improved BEND cell proliferation by enhancing PCNA and cyclin D1 expression via activation of the PI3K/AKT and MAPK signaling pathways. In addition, a combination of CCL23 and tunicamycin reversed the ER stress‐induced reduction in cell proliferation and the decreased expression of UPR‐mediated signaling proteins, including IRE1α, PERK, and ATF6α. Moreover, it regulated the lipopolysaccharide‐induced inflammation in BEND cells by inhibiting the expression of pro‐inflammatory cytokines (IL‐6 and IL‐8), and by restoring intracellular Ca2+ levels. These findings demonstrate that CCL23 improves endometrial development and uterine receptivity required for implantation and placentation during early pregnancy.In the present study, we identified that treatment of bovine endometrial (BEND) cells with CCL23 enhanced their proliferation with increased PCNA protein expression, and changed the number of cells within cell cycle phase via the PI3K/AKT and MAPK signal transduction pathways. Furthermore, the ER stress‐ and LPS‐induced reduction in cell proliferation was abolished by additional treatment of BEND cells with CCL23. These results provide evidence that CCL23 is essential for improving endometrial development and uterine receptivity required for implantation and placentation.
      PubDate: 2017-11-24T03:40:55.674932-05:
      DOI: 10.1002/jcp.26210
  • TIEG and estrogen modulate SOST expression in the murine skeleton
    • Authors: Malayannan Subramaniam; Kevin S. Pitel, Elizabeth S. Bruinsma, David G. Monroe, John R. Hawse
      Abstract: TIEG knockout (KO) mice exhibit a female‐specific osteopenic phenotype and altered expression of TIEG in humans is associated with osteoporosis. Gene expression profiling studies identified sclerostin as one of the most highly up‐regulated transcripts in the long bones of TIEG KO mice relative to WT littermates suggesting that TIEG may regulate SOST expression. TIEG was shown to substantially suppress SOST promoter activity and the regulatory elements through which TIEG functions were identified using promoter deletion and chromatin immunoprecipitation assays. Knockdown of TIEG in IDG‐SW3 osteocyte cells using shRNA and CRISPR‐Cas9 technology resulted in increased SOST expression and delayed mineralization, mimicking the results obtained from TIEG KO mouse bones. Given that TIEG is an estrogen regulated gene, and since changes in the hormonal milieu affect SOST expression, we performed ovariectomy (OVX) and estrogen replacement therapy (ERT) studies in WT and TIEG KO mice followed by miRNA and mRNA sequencing of cortical and trabecular compartments of femurs. SOST expression levels were considerably higher in cortical bone compared to trabecular bone. In cortical bone, SOST expression was increased following OVX only in WT mice and was suppressed following ERT in both genotypes. In contrast, SOST expression in trabecular bone was decreased following OVX and significantly increased following ERT. Interestingly, a number of miRNAs that are predicted to target sclerostin exhibited inverse expression levels in response to OVX and ERT. These data implicate important roles for TIEG and estrogen‐regulated miRNAs in modulating SOST expression in bone. This article is protected by copyright. All rights reserved
      PubDate: 2017-11-24T03:35:33.254612-05:
      DOI: 10.1002/jcp.26211
  • Effect of mono‐(2‐ethylhexyl) phthalate (MEHP) on proliferation of and
           steroid hormone synthesis in rat ovarian granulosa cells in vitro
    • Authors: Na Li; Te Liu, Kun Guo, Jian Zhu, Guangyan Yu, Shuyue Wang, Lin Ye
      Abstract: This study aimed to examine the proliferation of and secretion by rat ovarian granulosa cells (GCs) treated with mono‐(2‐ethylhexyl) phthalate (MEHP). Ovarian GCs were incubated with MEHP at concentration of 0, 25, 50, 100, and 200 µM for 24 hr. Cell viability was determined using the MTT Cell Proliferation Assay. Progesterone and estradiol production was evaluated by radioimmunoassay (RIA) and the expression of FSHR, PR, and ER was measured by immunocytochemistry. StAR, P450scc, 3β‐HSD, 17β‐HSD, and P450arom mRNA levels were determined by RT‐PCR. MEHP markedly attenuated proliferation of GCs, increased expression of sex hormone receptors and key enzymes in progesterone production, and stimulated steroid hormone secretion. The result of these analyses demonstrates that MEHP exposure of GCs may have effects on rat ovarian functions.The objectives of this work were to characterize mono‐(2‐ethylhexyl) phthalate (MEHP) cytotoxicity and effects on steroid hormone systems in rat ovarian granulosa cells (GCs) in vitro. Our results showed that MEHP inhibited the proliferation of GCs, increased sex hormone receptors and key enzymes in progesterone production, and caused abnormal steroid hormone synthesis, including increased progesterone and estradiol synthesis. Since MEHP is widely used in plastics, it has broad exposure potential in humans and should be studied further to establish the safety of this material.
      PubDate: 2017-11-24T03:30:32.478582-05:
      DOI: 10.1002/jcp.26224
    • Authors: Mattia Criscuoli; Irene Filippi, Daniela Osti, Carlo Aldinucci, Giuditta Guerrini, Giuliana Pelicci, Fabio Carraro, Antonella Naldini
      Abstract: Neuroblastoma (NB) is a highly malignant pediatric solid tumor where a hypoxic signature correlates with unfavorable patient outcome. The hypoxia‐inducible factor (HIF)‐1α plays an important role in NB progression, contributing to cell proliferation and invasiveness. RAI belongs to the Shc family proteins, it is mainly neuron specific and protects against cerebral ischemia. RAI is also expressed in several NB cell lines, where it promotes cell survival. In this work, hypoxia differently affected cell survival and pro‐apoptotic program in two NB cell lines, either expressing RAI (SKNBE) or not (SKNMC). RAI expression appeared to promote NB cell survival and to reduce some pro‐apoptotic markers under hypoxia. Accordingly, the RAI silencing in SKNBE cells resulted in a reduction of cell survival and HIF‐1α expression. Furthermore, using SKNMC cells stably expressing RAI, we defined a role of RAI in NB cell responses to hypoxia. Of interest, in hypoxic SKNMC cells expressing RAI HIF‐1α protein levels were higher than in control cells. This was associated with a) an increased cell survival; b) an increased expression of anti‐apoptotic markers; c) a pro‐autophagic and not pro‐apoptotic phenotype; d) an increased metabolic activity. We may conclude that RAI plays an important role in hypoxic signaling in NB cells and the interplay between RAI and HIF‐1α may be relevant in the protection of NB cells against hypoxia. Our results may contribute to a further understanding the physiology of NB cells and the molecular mechanisms involved in their survival, with important implications in NB progression. This article is protected by copyright. All rights reserved
      PubDate: 2017-11-24T03:26:08.28498-05:0
      DOI: 10.1002/jcp.26247
  • Effects of microRNA‐206 and its target gene IGF‐1 on
           sevoflurane‐induced activation of hippocampal astrocytes in aged rats
           through the PI3K/AKT/CREB signaling pathway
    • Authors: Tie-Jun Liu; Bin Wang, Qun-Xi Li, Xiao- Liu Dong, Xiao-Liang Han, Shu-Bo Zhang
      Abstract: The study aims to explore the effects of microRNA‐206 (miR‐206) targeting IGF‐1 on the activation of hippocampal astrocytes in aged rats induced by sevoflurane through the PI3K/AKT/CREB signaling pathway. Wistar rats and astrocytes were divided into the normal/blank, sham/negative control (NC), sevoflurane (sevo), miR‐206 mimics + sevo, miR‐206 inhibitors + sevo, miR‐206 NC + sevo, IGF‐1 shRNA + sevo and miR‐206 inhibitors + IGF‐1 shRNA + sevo groups. The Morris water maze test was exhibited to assess the cognitive functions. Glial fibrillary acidic protein (GFAP) expression was detected by immunofluorescence assay. Western blotting and RT‐qPCR were used to detect the expression of miR‐206, IGF‐1, PI3K, AKT, CREB, pPI3K, pAKT, pCREB, cytochrome‐c (Cyt‐c) and caspase‐3. Cell viability and apoptosis were detected by MTT assay and Annexin V/PI double staining respectively. Mitochondrial transmembrane potential (MTP) were determined by flow cytometry. The IGF‐1 shRNA + sevo group showed reduced miR‐206 expression. Compared with the normal/blank group, the sevo and miR‐206 NC + sevo groups showed decreased miR‐206 and GFAP expressions, cell viability and MTP but increased expressions of IGF‐1, PI3K, AKT, CREB, pPI3K, pAKT, pCREB, Cyt‐c and caspase‐3, as well as cell apoptosis. Similar trends were observed in the miR‐206 inhibitors + sevo group when compared with the sevo group. The study provides evidence that miR‐206 alleviates the inhibition of activation of hippocampal astrocytes in aged rats induced by sevoflurane by targeting IGT‐1 through suppressing the PI3K/AKT/CREB signaling pathway. This article is protected by copyright. All rights reserved
      PubDate: 2017-11-24T03:26:03.567827-05:
      DOI: 10.1002/jcp.26248
  • A high throughput method to study the physiology of E2:ERα signaling
           in breast cancer cells
    • Authors: Stefano Leone; Claudia Busonero, Filippo Acconcia
      Abstract: 17β‐estradiol (E2) regulates diverse physiological effects including cell proliferation through the estrogen receptor α (ERα), which as a transcription factor drives gene transcription and as an extra‐nuclear localized receptor triggers the membrane‐dependent activation of diverse kinase cascades. E2 also modifies ERα intracellular levels via diverse intracellular mechanisms. In this way, the E2‐acivated ERα integrates signaling cascades with the modulation of receptor intracellular concentration and with the induction of DNA synthesis and ultimately drives cell proliferation. In turn, E2 signaling deregulation can cause many diseases including breast cancer (BC). Recently, we performed a Western blotting (WB)‐based screen to identify novel pathways affecting ERα intracellular levels and BC cell proliferation. However, because WB lacks high throughput potential, a high‐content method to detect all aspects of E2:ERα signaling (nuclear and extra‐nuclear receptor activity, ERα levels, E2‐induced DNA synthesis) is desirable. Here, we set up a rapid way to measure E2:ERα signaling in 96‐well plate format. To demonstrate its robustness, we also challenged 4OH‐tamoxifen resistant (Tam‐Res) BC cells with a library of anti‐cancer drugs and identified methotrexate (MTX) as a molecule inducing ERα degradation and preventing BC cell proliferation. Overall, our research provides a high‐content technique to study the physiology of E2:ERα signaling in cells and further suggests a possible anti‐ERα and anti‐proliferative use for MTX in Tam‐Res BCs. This article is protected by copyright. All rights reserved
      PubDate: 2017-11-24T03:25:41.221019-05:
      DOI: 10.1002/jcp.26251
  • CpG oligodeoxynucleotide preconditioning improves cardiac function after
           myocardial infarction via modulation of energy metabolism and angiogenesis
    • Authors: Deng-Cheng Zhou; Yong-Hui Su, Fu-Qing Jiang, Jing-Bo Xia, Hai-Yan Wu, Zao-Shang Chang, Wen-Tao Peng, Guo-Hua Song, Kyu-Sang Park, Soo-Ki Kim, Dong-Qing Cai, Li Zheng, Xu-Feng Qi
      Abstract: Unmethylated CpG oligodeoxynucleotide (CpG‐ODN), a Toll‐like receptor 9 (TLR9) ligand, has been shown to protect against myocardial ischemia/reperfusion injury. However, the potential effects of CpG‐ODN on myocardial infarction (MI) induced by persistent ischemia remains unclear. Here, we investigated whether and how CpG‐ODN preconditioning protects against MI in mice. C57BL/6 mice were treated with CpG‐ODN by i.p. injection 2 hours prior to MI induction, and cardiac function and histology were analyzed two weeks after MI. Both 1826‐CpG and KSK‐CpG preconditioning significantly improved the left ventricular (LV) ejection fraction (LVEF) and LV fractional shortening (LVFS) when compared with non‐CpG controls. Histological analysis further confirmed the cardioprotection of CpG‐ODN preconditioning. In vitro studies further demonstrated that CpG‐ODN preconditioning increases cardiomyocyte survival under hypoxic/ischemic conditions by enhancing stress tolerance through TLR9‐mediated inhibition of the SERCA2/ATP and activation of AMPK pathways. Moreover, CpG‐ODN preconditioning significantly increased angiogenesis in the infarcted myocardium compared with non‐CpG. However, persistent TLR9 activation mediated by lentiviral infection failed to improve cardiac function after MI. Although CpG‐ODN preconditioning increased angiogenesis in vitro, both the persistent stimulation of CpG‐ODN and stable overexpression of TLR9 suppressed the tube formation of cardiac microvascular endothelial cells. CpG‐ODN preconditioning significantly protects cardiac function against MI by suppressing the energy metabolism of cardiomyocytes and promoting angiogenesis. Our data also indicate that CpG‐ODN preconditioning may be useful in MI therapy. This article is protected by copyright. All rights reserved
      PubDate: 2017-11-24T03:18:40.825128-05:
      DOI: 10.1002/jcp.26243
  • Transcription factor HMG box‐containing protein 1 (HBP1) modulates
           mitotic clonal expansion (MCE) during adipocyte differentiation
    • Authors: Chien-Yi Chan; Ping Yu, Feng-Tzu Chang, Zih-Hua Chen, Ming-Fen Lee, Chun-Yin Huang
      Abstract: Transcription factor HMG box‐containing protein 1 (HBP1) has been found to be up‐regulated in rat adipose tissue and differentiated preadipocyte; however, how HBP1 is involved in adipocyte formation remains unclear. In the present study, we demonstrated that under a standard differentiation protocol HBP1 expression fluctuates with down‐regulation in the mitotic clonal expansion (MCE) stage followed by up‐regulation in the terminal differentiation stage in both 3T3‐L1 and MEF cell models. Also, HBP1 knockdown accelerated cell cycle progression in the MCE stage, but it impaired final adipogenesis. To gain further insight into the role of HBP1 in the MCE stage, we found that the HBP1 expression pattern is reciprocal to that of C/EBPβ, and ectopic expression of HBP1suppresses C/EBPβ expression. These data indicate that HBP1 functions as a negative regulator of MCE. In contrast, when HBP1 expression was gradually elevated along with a concomitant induction of C/EBPα at the end of the MCE, HBP1 knockdown leads to a significant reduction of C/EBPα expression, suggesting that HBP1‐mediated C/EBPα expression may be needed for the termination of the cell cycle at the end of MCE for terminal differentiation. All told, our findings show that HBP1 is a key transcription factor in the already complicated regulatory cascade during adipocyte differentiation.HBP1 regulates adipocyte differentiation. Upon MDI stimulation, HBP1 expression is down‐regulated at the beginning of the MCE to ensure the up‐regulation of C/EBPβ and allows the growth‐arrested preadipocytes to reentry into postconfluent mitosis. However, HBP1 expression is up‐regulated at the end of the MCE to ensure the up‐regulation of C/EBPα and the termination of the MCE as well as the onset of terminal differentiation.
      PubDate: 2017-11-24T03:18:20.903688-05:
      DOI: 10.1002/jcp.26237
  • Clinical and Prognostic Value of the C‐Met/HGF Signaling Pathway in
           Cervical Cancer
    • Authors: Nadia Boromand; Malihe Hasanzadeh, Soodabeh ShahidSales, Marjaneh Farazestanian, Masoumeh Gharib, Hamid Fiuji, Negin Behboodi, Niloofar Ghobadi, Seyed Mahdi Hassanian, Gordon A. Ferns, Amir Avan
      Abstract: Aberrant activation of the HGF/c‐Met signalling pathway is reported to be associated with cell proliferation, progression, and metastasis features of several tumor types, including cervical cancer, suggesting that it may be of potential value as a novel therapeutic target. Furthermore, HPV‐positive patients had a higher serum level of HGF or c‐Met protein, compared with HPV‐negative patients. c‐Met or HGF overexpression in lesions of cervical cancer is reported to be related to a poorer prognosis, and hence this may be of value as a prognostic and predictive biomarker. Several approaches have been developed for targeting HGF and/or c‐Met. One of these is crizotinib (a dual c‐Met/ALK inhibitor). This has been approved by FDA for the treatment of lung‐cancer. Further investigations are required to evaluate and optimize the use of c‐Met inhibitors in cervical cancer or parallel targeting signalling pathway associated/activated via MET/HGF pathway. The main aim of current review was to give an overview of the potential of the c‐Met/HGF pathway as a prognostic, or predictive biomarker in cervical cancer. This article isprotected by copyright. All rights reserved
      PubDate: 2017-11-24T02:20:21.061138-05:
      DOI: 10.1002/jcp.26232
  • Lipid signaling affects primary fibroblast collective migration and
           anchorage in response to stiffness and microtopography
    • Authors: Michael A. Mkrtschjan; Snehal B. Gaikwad, Kevin J. Kappenman, Christopher Solís, Sagar Dommaraju, Long V. Le, Tejal A. Desai, Brenda Russell
      Abstract: Cell migration is regulated by several mechanotransduction pathways, which consist of sensing and converting mechanical microenvironmental cues to internal biochemical cellular signals, such as protein phosphorylation and lipid signaling. While there has been significant progress in understanding protein changes in the context of mechanotransduction, lipid signaling is more difficult to investigate. In this study, physical cues of stiffness (10 kPa, 100 kPa, 400 kPa, and glass), and microrod or micropost topography were manipulated in order to reprogram primary fibroblasts and assess the effects of lipid signaling on the actin cytoskeleton. In an in vitro wound closure assay, primary cardiac fibroblast migration velocity was significantly higher on soft polymeric substrata. Modulation of PIP2 availability through neomycin treatment nearly doubled migration velocity on 10 kPa substrata, with significant increases on all stiffnesses. The distance between focal adhesions and the lamellar membrane (using wortmannin treatment to increase PIP2 via PI3K inhibition) was significantly shortest compared to untreated fibroblasts grown on the same surface. PIP2 localized to the leading edge of migrating fibroblasts more prominently in neomycin‐treated cells. The membrane‐bound protein, lamellipodin, did not vary under any condition. Additionally, fifteen micron‐high micropost topography, which blocks migration, concentrates PIP2 near to the post. Actin dynamics within stress fibers, measured by fluorescence recovery after photobleaching, was not significantly different with stiffness, microtopography, nor with drug treatment. PIP2‐modulating drugs delivered from microrod structures also affected migration velocity. Thus, manipulation of the microenvironment and lipid signaling regulatory drugs might be beneficial in improving therapeutics geared toward wound healing. This article is protected by copyright. All rights reserved
      PubDate: 2017-11-24T02:15:48.957032-05:
      DOI: 10.1002/jcp.26236
  • Regulating the transcriptomes that mediate the conversion of fibroblasts
           to various nervous system neural cell types
    • Authors: Niusha Khazaei; Shima Rastegar-Pouyani, Nicholas O'Toole, Ping Wee, Abdulshakour Mohammadnia, Moein Yaqubi
      Abstract: Our understanding of the mechanism of cell fate transition during the direct reprogramming of fibroblasts into various central nervous system (CNS) neural cell types has been limited by the lack of a comprehensive analysis on generated cells, independently and in comparison with other CNS neural cell types. Here, we applied an integrative approach on 18 independent high throughput expression data sets to gain insight into the regulation of the transcriptome during the conversion of fibroblasts into induced neural stem cells, induced neurons, induced astrocytes, and induced oligodendrocyte progenitor cells. We found common down‐regulated genes to be mostly related to fibroblast‐specific functions, and suggest their potential as markers for screening of the silencing of the fibroblast‐specific program. For example, Tagln was significantly down‐regulated across all considered data sets. In addition, we identified specific profiles of up‐regulated genes for each CNS neural cell types, which could be potential markers for maturation and efficiency screenings. Furthermore, we identified the main TFs involved in the regulation of the gene expression program during direct reprogramming. For example, in the generation of induced neurons from fibroblasts, the Rest TF was the main regulator of this reprogramming. In summary, our computational approach for meta‐analyzing independent expression data sets provides significant details regarding the molecular mechanisms underlying the regulation of the gene expression program, and also suggests potentially useful candidate genes for screening down‐regulation of fibroblast gene expression profile, maturation, and efficiency, as well as candidate TFs for increasing the efficiency of the reprogramming process. This article is protected by copyright. All rights reserved
      PubDate: 2017-11-24T02:10:26.957294-05:
      DOI: 10.1002/jcp.26221
  • Melatonin application in targeting oxidative‐induced liver injuries:
           A review
    • Authors: Keywan Mortezaee; Neda Khanlarkhani
      Abstract: It is believed that oxidative stress is a key causing factor of liver damage induced by a variety of agents, and it is a major contributing factor in almost all conditions compromising liver function, including ischemia–reperfusion injury (IRI), nonalcoholic fatty liver disease (NAFLD), nonalcoholic steatohepatitis (NASH), liver fibrosis, liver cirrhosis, and hepatocellular carcinoma (HCC). Liver is the organ that high concentration of melatonin (N‐acetyl‐5‐methoxytryptamine) accumulates, and it is the sole organ where circulating melatonin is metabolized. Melatonin is one of the best antioxidants that protects liver, and its metabolites also have antioxidative function. Melatonin exerts its antioxidative function directly through its radical scavenging ability and indirectly through stimulation of antioxidant enzymes. The antioxidative response from melatonin in liver affects from various factors, including its dosage, route, time and duration of administration, the type of oxidative‐induced agent and species aging. This indoleamine is also an effective and promising antioxidative choice for targeting liver IRI, NAFLD, NASH, fibrosis, cirrhosis, and HCC.Melatonin is one of the best antioxidants that protects liver. Melatonin exerts its antioxidative function directly through its radical scavenging ability and indirectly through stimulation of antioxidant enzymes. This indoleamine is also an effective and promising antioxidative choice for targeting liver IRI, NAFLD, NASH, fibrosis, cirrhosis, and HCC.
      PubDate: 2017-11-24T02:01:12.680204-05:
      DOI: 10.1002/jcp.26209
  • miR‐19b‐3p induces cell proliferation and reduces
           heterochromatin‐mediated senescence through PLZF in goat male germline
           stem cells
    • Authors: John Clotaire Daguia Zambe; Yuanxin Zhai, Zhe Zhou, Xiaomi Du, Yudong Wei, Fanglin Ma, Jinlian Hua
      Abstract: Promyelocytic leukemia zinc finger PLZF, known as ZBTB16 or ZFP145 is a critical zinc finger protein of male germline stem cells (mGSCs), it's an essential transcriptional factor for goat testis development and spermatogenesis. Loss of PLZF results in progressive depletion of SSCs after the first wave of spermatogenesis leading to eventual spermatogenic arrest, apparently the result of a shift in the balance in SSC fate away from self‐renewal and toward differentiation. Cumulating evidences have demonstrated that microRNAs are expressed in a cell‐specific or stage‐specific manner during spermatogenesis and acts as regulators on specific makers such as Stra8, ETV5, and PLZF. However, the post transcriptional function of PLZF still poorly elucidate in mGSCs. Bioinformatic analysis and dual luciferase reporter assay showed that miR‐19b‐3p binds the 3′UTR of PLZF, suggesting that PLZF is a direct target of miR‐19b‐3p. The profile of miR‐19b‐3p and PLZF analyzed in dairy goat testis at different age showed that miR‐19b‐3p was significantly up‐regulated in goat testis at 1, 3, 6 months and downregulated at 12, 18, and 24 months which was inversely correlated with PLZF in the same testis. Focusing on the role of miR‐19b‐3p, we found that miR‐19b‐3p changes c‐KIT and mTOR signaling through PLZF to promote proliferation in goat nGSCs and infertile mice testes. Over‐expression of PLZF significantly reversed miR‐19b‐3p‐mediated proliferation in mice testes. We found also that miR‐19b‐3p reduced heterochromatin‐mediated senescence through PLZF localized on HP1α. Taken together, our findings indicate that miR‐19b‐3p promotes proliferation and reduces heterochromatin‐mediated senescence through PLZF in mGSCs.miR‐19b‐3p changes c‐KIT and mTOR signaling through PLZF, induces goat mGSCs proliferation and reduces heterochromatin‐mediated cell senescence.
      PubDate: 2017-11-24T02:01:05.197444-05:
      DOI: 10.1002/jcp.26231
  • Differences in definitive endoderm induction approaches using growth
           factors and small molecules
    • Authors: Mariia S. Bogacheva; Sofia Khan, Liisa K. Kanninen, Marjo Yliperttula, Alan W. Leung, Yan-Ru Lou
      Abstract: Definitive endoderm (DE) is the first stage of human pluripotent stem cell (hPSC) differentiation into hepatocyte‐like cells. Developing human liver cell models for pharmaceutical applications is highly demanding. Due to the vast number of existing protocols to generate DE cells from hPSCs, we aimed to compare the specificity and efficiency of selected published differentiation conditions. We differentiated two hPSC lines (induced PSC and embryonic stem cell) to DE cells on Matrigel matrix using growth factors (Activin A and Wnt‐3a) and small molecules (sodium butyrate and IDE 1) in different combinations. By studying dynamic changes during six days in cell morphology and the expression of markers for pluripotency, DE, and other germ layer lineages, we found that Activin A is essential for DE differentiation, while Wnt‐3a and sodium butyrate are dispensable. Although sodium butyrate exerted rapid DE differentiation kinetics, it caused massive cell death and could not generate sufficient cells for further differentiation and applications. We further discover that IDE 1 could not induce DE as reported previously. Hereby, we compared different conditions for DE induction and found an effective six day‐protocol to obtain DE cells for the further differentiation and applications. This article is protected by copyright. All rights reserved
      PubDate: 2017-11-23T06:31:01.383803-05:
      DOI: 10.1002/jcp.26214
  • MiR‐145‐5p inhibits proliferation and inflammatory responses of RMC
           through regulating AKT/GSK pathway by targeting CXCL16
    • Authors: Junbiao Wu; Yu He, Yining Luo, Lei Zhang, Hua Lin, Xusheng Liu, Bihao Liu, Chunling Liang, Yuan Zhou, Jiuyao Zhou
      Abstract: The main pathological characteristics of chronic glomerulonephritis (CGN) are diffuse mesangial cells proliferation and inflammatory responses. Our previous studies have confirmed that miR‐145‐5p was abnormally elevated in CGN rats, but its mechanism remains unclear. Therefore, this study aimed to elucidate the mechanism of miR‐145‐5p in regulation of renal mesangial cells proliferation and inflammatory responses. In vivo study, the cationic bovine serum albumin (C‐BSA)‐induced CGN rat model was established, and the content of miR‐145‐5p in renal was examined by qRT‐PCR, meanwhile, we also determined the renal function and inflammatory infiltrate. In vitro, the cell proliferation rate, cell cycle and inflammatory changes of rat mesangial cells (RMCs) were measured. Our results suggested that miR‐145‐5p extended the G0‐G1 phase, shortened S phase, inhibited cell proliferation and suppressed inflammatory responses in RMCs. Moreover, miR‐145‐5p inhibited CXCL16 protein expression through binding the 3′‐UTR of CXCL16, suppressed AKT/GSK signaling pathway, and decreased expression of inflammation related mRNAs, such as IL‐1α, IL‐2, IL‐6, and TNF‐α mRNAs. Further, locking CXCL16 alleviated inflammatory reactions and down‐regulated AKT/GSK pathway in RMCs. Above all, we concluded that miR‐145‐5p inhibited proliferation and inflammatory responses of RMCs through regulation of AKT/GSK pathway by targeting CXCL16.Our study aimed to elucidate the mechanism of miR‐145‐5p in regulation of renal mesangial cell proliferation and inflammatory responses. Base on the in vivo and in vitro study, the results suggested that miR‐145‐5p inhibited proliferation and inflammatory responses of RMCs through regulation of AKT/GSK pathway by targeting CXCL16.
      PubDate: 2017-11-23T06:15:55.543253-05:
      DOI: 10.1002/jcp.26228
  • Extracellular adenosine‐induced Rac1 activation in pulmonary
           endothelium: molecular mechanisms and barrier‐protective role
    • Authors: Anita Kovacs-Kasa; Kyung Mi Kim, Mary Cherian-Shaw, Stephen M. Black, David J. Fulton, Alexander D. Verin
      Abstract: We have previously shown that Gs‐coupled adenosine receptors (A2a) are primarily involved in adenosine‐induced human pulmonary artery endothelial cell (HPAEC) barrier enhancement. However, the downstream events that mediate the strengthening of the endothelial cell (EC) barrier via adenosine signaling are largely unknown. In the current study we tested the overall hypothesis that adenosine‐induced Rac1 activation and EC barrier enhancement is mediated by Gs‐dependent stimulation of cAMP‐dependent Epac1‐mediated signaling cascades. Adenoviral transduction of HPAEC with constitutively‐active (C/A) Rac1 (V12Rac1) significantly increases transendothelial electrical resistance (TER) reflecting an enhancement of the EC barrier. Conversely, expression of an inactive Rac1 mutant (N17Rac1) decreases TER reflecting a compromised EC barrier. The adenosine‐induced increase in TER was accompanied by activation of Rac1, decrease in contractility (MLC dephosphorylation), but not Rho inhibition. Conversely, inhibition of Rac1 activity attenuates adenosine‐induced increase in TER. We next examined the role of cAMP‐activated Epac1 and its putative downstream targets Rac1, Vav2, Rap1 and Tiam1. Depletion of Epac1 attenuated the adenosine‐induced Rac1 activation and the increase in TER. Furthermore, silencing of Rac1 specific guanine nucleotide exchange factors (GEFs), Vav2 and Rap1a expression significantly attenuated adenosine‐induced increases in TER and activation of Rac1. Depletion of Rap1b only modestly impacted adenosine‐induced increases in TER and Tiam1 depletion had no effect on adenosine‐induced Rac1 activation and TER. Together these data strongly suggest that Rac1 activity is required for adenosine‐induced EC barrier enhancement and that the activation of Rac1 and ability to strengthen the EC barrier depends, at least in part, on cAMP‐dependent Epac1/Vav2/Rap1‐mediated signaling. This article is protected by copyright. All rights reserved
      PubDate: 2017-11-23T00:50:35.42441-05:0
      DOI: 10.1002/jcp.26281
  • Therapeutic potency of mTOR signaling pharmacological inhibitors in the
           treatment of proinflammatory diseases, current status and perspectives
    • Authors: Arash Soltani; Amirhossein Bahreyni, Nadia Boroumand, Mostafa karimi Roshan, Majid Khazaei, Mikhail Ryzhikov, Saman Soleimanpour, Amir Avan, Seyed Mahdi Hassanian
      Abstract: Mammalian target of rapamycin (mTOR) signaling pathway controls cell energy metabolism. There is an interplay between mTOR and proinflammatory signaling pathways, supporting the role of the pathway in the pathogenesis of inflammatory diseases. Inhibition of mTOR signaling using specific pharmacological inhibitors could offer therapeutic promise in several inflammatory‐associated diseases. In this review, we summarize recent findings on the regulatory effects of mTOR signaling on inflammation and the therapeutic potency of mTOR pharmacological inhibitors in the treatment of inflammatory diseases including cancer, neurodegenerative diseases, atherosclerosis, sepsis and rheumatoid arthritis for a better understanding and hence a better management of these diseases. This article is protected by copyright. All rights reserved
      PubDate: 2017-11-22T06:55:20.195488-05:
      DOI: 10.1002/jcp.26276
  • Pharmacological and molecular approaches for the treatment of
           β‐hemoglobin disorders
    • Authors: Neelam Lohani; Nupur Bhargava, Anjana Munshi, Sivaprakash Ramalingam
      Abstract: β‐hemoglobin disorders, such as β‐thalassemia and sickle cell anemia are among the most prevalent inherited genetic disorders worldwide. These disorders are caused by mutations in the gene encoding hemoglobin‐β (HBB), a vital protein found in red blood cells (RBCs) that carries oxygen from lungs to all parts of the human body. As a consequence, there has been an enduring interest in this field in formulating therapeutic strategies for the treatment of these diseases. Currently, there is no cure available for hemoglobin disorders, although, some patients have been treated with bone marrow transplantation, whose scope is limited because of the difficulty in finding a histocompatible donor and also due to transplant‐associated clinical complications that can arise during the treatment. On account of these constraints, reactivation of fetal hemoglobin (HbF) synthesis holds immense promise and is a viable strategy to alleviate the symptoms of β‐hemoglobin disorders. Development of new genomic tools has led to the identification of important natural genetic modifiers of hemoglobin switching which include BCL11A, KLF1, HBSIL‐MYB, LRF, LSD1, LDB1, histone deacetylases 1 and 2 (HDAC1 and HDAC2). miRNAs are also promising therapeutic targets for development of more effective strategies for the induction of HbF production. Many new small molecule pharmacological inducers of HbF production are already under pre‐clinical and clinical development. Furthermore, recent advancements in gene and cell therapy includes targeted genome editing and iPS cell technologies, both of which utilizes a patient's own cells, are emerging as extremely promising approaches for significantly reducing the burden of β‐hemoglobin disorders. This article is protected by copyright. All rights reserved
      PubDate: 2017-11-20T22:40:31.456005-05:
      DOI: 10.1002/jcp.26292
    • Authors: Francesca Veronesi; Veronica Borsari, Maria Sartori, Monia Orciani, Monica Mattioli-Belmonte Cima, Milena Fini
      Abstract: Tissue regenerative medicine combines the use of cells, scaffolds and molecules to repair damaged tissues. Different cell types are employed for musculoskeletal diseases, both differentiated and mesenchymal stromal cells (MSCs). In recent years, the hypothesis that cell‐based therapy is guided principally by cell‐secreted factors has become increasingly popular. The aim of the present literature review was to evaluate preclinical and clinical studies that used conditioned medium (CM), rich in cell‐factors, for musculoskeletal regeneration with a cell‐free approach. Thirty‐one were in vitro, 12 in vivo studies, 1 was a clinical study and 2 regarded extracellular vesicles. Both differentiated cells and MSCs produce CM that induces reduction in inflammation and increases synthetic activity. MSC recruitment and differentiation, endothelial cell recruitment and angiogenesis have also been observed. In vivo studies were performed with CM in bone and periodontal defects, arthritis and muscle dystrophy pathologies. The only clinical study was performed with CM from MSCs in patients needing alveolar bone regeneration, showing bone formation and no systemic or local complications. Platelet derived growth factor receptor β, C3a, vascular endothelial growth factor, monocyte chemoattractant protein‐1 and ‐3, interleukin 3 and 6, insulin‐like growth factor‐I were identified as responsible of cell migration, proliferation, osteogenic differentiation and angiogenesis.The use of CM could represent a new regenerative treatment in several musculoskeletal pathologies because it overcomes problems associated with the use of cells and avoids the use of exogenous GFs or gene delivery systems. However, some issues remain to be clarified. This article is protected by copyright. All rights reserved
      PubDate: 2017-11-20T22:40:29.389255-05:
      DOI: 10.1002/jcp.26291
  • Inhibition of HSP90α protects cultured neurons from oxygen‐glucose
           deprivation induced necroptosis by decreasing RIP3 expression
    • Authors: Zhen Wang; Li-min Guo, Yong Wang, Hong-kang Zhou, Shu-chao Wang, Dan Chen, Ju-fang Huang, Kun Xiong
      Abstract: Heat shock protein 90α (HSP90α) maintains cell stabilization and regulates cell death, respectively. Recent studies have shown that HSP90α is involved in receptor interacting protein 3 (RIP3)‐mediated necroptosis in HT29 cells. It is known that oxygen and glucose deprivation (OGD) can induce necroptosis, which is regulated by RIP3 in neurons. However, it is still unclear whether HSP90α participates in the process of OGD‐induced necroptosis in cultured neurons via the regulation of RIP3. Our study found that necroptosis occurs in primary cultured cortical neurons and PC‐12 cells following exposure to OGD insult. Additionally, the expression of RIP3/p‐RIP3, MLKL/p‐MLKL, and the RIP1/RIP3 complex (necrosome) significantly increased following OGD, as measured through immunofluorescence (IF) staining, western blotting (WB), and immunoprecipitation (IP) assay. Additionally, data from computer simulations and IP assays showed that HSP90α interacts with RIP3. In addition, HSP90α was overexpressed following OGD in cultured neurons, as measured through WB and IF staining. Inhibition of HSP90α in cultured neurons, using the specific inhibitor, geldanamycin (GA), and siRNA/shRNA of HSP90α, protected cultured neurons from necrosis. Our study showed that the inhibitor of HSP90α, GA, rescued cultured neurons not only by decreasing the expression of total RIP3/MLKL, but also by decreasing the expression of p‐RIP3/p‐MLKL and the RIP1/RIP3 necrosome. In this study, we reveal that inhibition of HSP90α protects primary cultured cortical neurons and PC‐12 cells from OGD‐induced necroptosis through the modulation of RIP3 expression. This article is protected by copyright. All rights reserved
      PubDate: 2017-11-20T08:36:08.701548-05:
      DOI: 10.1002/jcp.26294
  • Baicalein Inhibits Acinar‐to‐Ductal Metaplasia of Pancreatic Acinal
           Cell AR42J via Improving the Inflammatory Microenvironment
    • Authors: Wei-ling PU; Ying-ying LUO, Ru-Yu BAI, Ao-wei GUO, Kun ZHOU, Yun-sha ZHANG, Lin MIAO, Curzio RÜEGG, Micheal O. HOTTIGER, Xiu-mei GAO, Li-kang SUN
      Abstract: Pancreatic ductal adenocarcinoma (PDAC) is one of the most aggressive cancers. Recent research has demonstrated that chronic pancreatitis (CP) is associated with an increased risk of PDAC, partly due to acinar‐to‐ductal metaplasia (ADM). Baicalein has been shown to exert anti‐inflammatory and anti‐tumor effects for CP or PDAC, respectively. The aim of our study was to investigate the effect of baicalein, and the putative underlying mechanism, on inflammatory cytokines‐induced ADM of rat pancreatic acinar cell line AR42J. To investigate ADM and baicalein effects in vitro, AR42J were treated with recombinant rat Tumor Necrosis Factor alpha (rTNFα) with or without baicalein for 5 days. Results showed that rTNFα‐induced AR42J cells switched their phenotype from dominantly amylase‐positive acinar cells to dominantly cytokeratin 19‐positive ductal cells. Moreover, expression of the transcripts for TNFα or Hes‐1, a Notch target, was up‐regulated in these cells. Interestingly, baicalein reduced the population of ADM as well as cytokines gene expression but not Hes‐1. Baicalein inhibited NF‐κB activation induced by rTNFα in AR42J, but no effect on Notch 1activation. Moreover, baicalein suppressed the secretion of TNFα and Nitric Oxide (NO) in macrophages stimulated with LPS and further inhibited ADM of conditional medium‐treated AR42J cells. Baicalein also suppressed the inflammatory response of LPS‐activated macrophages, thereby inhibited ADM of AR42J by altering their microenvironment. Taken together, our study indicates that baicalein reduces rTNFα‐induced ADM of AR42J cells by inhibiting NF‐κB activation. It also sheds new light on Chinese material medica therapy of pancreatitis and thereby prevention of PDAC. This article is protected by copyright. All rights reserved
      PubDate: 2017-11-20T08:30:23.77799-05:0
      DOI: 10.1002/jcp.26293
    • Authors: Simona Ultimo; Alberto M. Martelli, Giorgio Zauli, Marco Vitale, George A. Calin, Luca M. Neri
      Abstract: MicroRNAs (miRNAs) are a class of small noncoding RNAs which regulate the expression of target genes by binding to messenger RNAs. miRNAs play a role in various biological processes, including proliferation, apoptosis and tumorigenesis. Dysregulation of miRNAs is implicated in invasion and metastasis in several human cancer types, and leukemia is not an exception.Acute Lymphoblastic Leukemia (ALL) is an hematological malignancy characterized by the proliferation of early lymphoid precursors that replace normal hematopoietic cells of the bone marrow. The expression profiling of miRNAs in ALL could be used for the classification of the disease establishing specific diagnoses and offering prognostic values in the near future. The correlation of miRNAs dysregulation and biology of ALL demonstrates that specific miRNA may be a potential therapy target. In this review we have focused our attention on the correlations between ALL and miRNAs, their link with signaling pathways and transcription factors in the disease and miRNA targeting therapeutic strategies with their advantages and potential use in clinical applications. This article is protected by copyright. All rights reserved
      PubDate: 2017-11-20T08:25:35.889233-05:
      DOI: 10.1002/jcp.26290
  • ΔFosB regulates rosiglitazone‐induced milk fat synthesis and
           cell survival
    • Authors: Xuefeng Wei; Hui Li, Guangwei Zhao, Jiameng Yang, Lihui Li, Yongzhen Huang, Xianyong Lan, Yun Ma, Huiling Zheng, Hong Chen
      Abstract: Rosiglitazone induces adipogenesis in adipocyte and regulates cell survival and differentiation in number of cell types. However, whether PPARγ regulates the synthesis of milk fat and cell survival in goat mammary gland remains unknown. Rosiglitazone strongly enhanced cellular triacylglycerol content and accumulation of lipid droplet in goat mammary epithelial cells (GMEC). Furthermore, ΔFosB decreased the expression of PPARγ at both mRNA and protein levels, and rosiglitazone‐induced milk fat synthesis was abolished by ΔFosB overexpression. ΔFosB reduced milk fat synthesis and enhanced saturated fatty acid concentration. Rosiglitazone increased the number of GMEC in G0/G1 phase and inhibited cell proliferation, and these effects were improved by overexpression of ΔFosB. ΔFosB was found to promote the expression of Bcl‐2 and suppress the expression of Bax, and protected GMEC from apoptosis induced by rosiglitazone. Intracellular calcium trafficking assay revealed that rosiglitazone markedly increased intracellular calcium concentration. ΔFosB protected GMEC from apoptosis induced by intracellular Ca2+ overload. ΔFosB increased MMP‐9 gelatinolytic activity. SB‐3CT, an MMP‐9 inhibitor, suppressed the expression of Bcl‐2 and increased intracellular calcium levels, and this effect was abolished by ΔFosB overexpression. SB‐3CT induced GMEC apoptosis and this effect was inhibited by ΔFosB overexpression. These findings suggest that ΔFosB regulates rosiglitazone‐induced milk fat synthesis and cell survival. Therefore, ΔFosB may be an important checkpoint to control milk fat synthesis and cell apoptosis. This article is protected by copyright. All rights reserved
      PubDate: 2017-11-20T08:25:24.820156-05:
      DOI: 10.1002/jcp.26218
  • Long non‐coding RNA MALAT1 promotes oral squamous cell carcinoma
           development via microRNA‐125b/STAT3 axis
    • Authors: Shi-Min Chang; Wei-Wei Hu
      Abstract: Oral squamous cell carcinoma (OSCC), as the most common type of oral cancer, is responsible for almost 3% of all malignant tumors worldwide. Non‐coding RNAs such as lncRNAs and microRNAs have been involved in many cancers including OSCC. Recently, lncRNA metastasis‐associated lung adenocarcinoma transcript‐1 (MALAT1) has been reported to play an oncogenic role in OSCC metastasis. However, the underlying mechanism of MALAT1 in regulating OSCC progression remains unclear. The aim of this study was to investigate the specific role of MALAT1 in OSCC development. It was observed that MALAT1 was upregulated in OSCC cell lines. Inhibition of MALAT1 can prevent OSCC proliferation while overexpressing MALAT1 promoted OSCC progression. In addition, bioinformatics search was used to identify that miR‐125b was a direct target of MALAT1, which indicated a negative correlation between MALAT1 and miR‐125b. Besides these, STAT3 was predicted as a binding target of miR‐125b in OSCC. Overexpression of MALAT1 was able to suppress the tumor inhibitory effect of miR‐125b mimics via upregulating STAT3. Moreover, the function of MALAT1 in OSCC development was further investigated by using in vivo assays. The established nude mice models revealed that downregulated MALAT1 greatly inhibited OSCC tumor growth and reversely upregualated MALAT1 promoted OSCC development via miR‐125b/STAT3 axis respectively. In conclusion, MALAT1 can function as a competing endogenous RNA (ceRNA) to modulate STAT3 expression by absorbing miR‐125b in OSCC and could be used as a novel therapeutic target in OSCC diagnosis and treatment. This article is protected by copyright. All rights reserved
      PubDate: 2017-11-20T07:15:27.172187-05:
      DOI: 10.1002/jcp.26185
  • Comprehensive transcriptome analysis of fluid shear stress altered gene
           expression in renal epithelial cells
    • Authors: Steven J. Kunnen; Tareq B. Malas, Cornelis M. Semeins, Astrid D. Bakker, Dorien J. M. Peters
      Abstract: Renal epithelial cells are exposed to mechanical forces due to flow‐induced shear stress within the nephrons. Shear stress is altered in renal diseases caused by tubular dilation, obstruction and hyperfiltration, which occur to compensate for lost nephrons. Fundamental in regulation of shear stress are primary cilia and other mechano‐sensors, and defects in cilia formation and function have profound effects on development and physiology of kidneys and other organs. We applied RNA sequencing to get a comprehensive overview of fluid‐shear regulated genes and pathways in renal epithelial cells. Functional enrichment‐analysis revealed TGF‐β, MAPK and Wnt signaling as core signaling pathways up‐regulated by shear. Inhibitors of TGF‐β and MAPK/ERK signaling modulate a wide range of mechanosensitive genes, identifying these pathways as master regulators of shear‐induced gene expression. However, the main down‐regulated pathway, i.e. JAK/STAT, is independent of TGF‐β and MAPK/ERK. Other up‐regulated cytokine pathways include FGF, HB‐EGF, PDGF and CXC. Cellular responses to shear are modified at several levels, indicated by altered expression of genes involved in cell‐matrix, cytoskeleton and glycocalyx remodeling, as well as glycolysis and cholesterol metabolism. Cilia ablation abolished shear induced expression of a subset of genes, but genes involved in TGF‐β, MAPK and Wnt signaling were hardly affected, suggesting that other mechano‐sensors play a prominent role in the shear stress response of renal epithelial cells. Modulations in signaling due to variations in fluid shear stress are relevant for renal physiology and pathology, as suggested by elevated gene expression at pathological levels of shear stress compared to physiological shear. This article is protected by copyright. All rights reserved
      PubDate: 2017-11-20T07:10:54.024502-05:
      DOI: 10.1002/jcp.26222
  • Intracrine prostaglandin E2 pro‐tumoral actions in prostate epithelial
           cells originate from non‐canonical pathways
    • Authors: Antonio Madrigal-Martínez; Ana B. Fernández-Martínez, Francisco J. Lucio Cazaña
      Abstract: Prostaglandin E2 (PGE2) increases cell proliferation and stimulates migratory and angiogenic abilities in prostate cancer cells. However, the effects of PGE2 on non‐transformed prostate epithelial cells are unknown, despite the fact that PGE2 overproduction has been found in benign hyperplastic prostates. In the present work we studied the effects of PGE2 in immortalized, non‐malignant prostate epithelial RWPE‐1 cells and found that PGE2 increased cell proliferation, cell migration, and production of vascular endothelial growth factor‐A, and activated in vitro angiogenesis. These actions involved a non‐canonic intracrine mechanism in which the actual effector was intracellular PGE2 (iPGE2) instead of extracellular PGE2: inhibition of the prostaglandin uptake transporter (PGT) or antagonism of EP receptors prevented the effects of PGE2, which indicated that PGE2 activity depended on its carrier‐mediated translocation from the outside to the inside of cells and that EP receptors located intracellularly (iEP) mediated the effects of PGE2. iPGE2 acted through transactivation of epidermal growth factor‐receptor (EGFR) by iEP, leading to increased expression and activity of hypoxia‐inducible factor‐1α (HIF‐1α). Interestingly, iPGE2 also mediates the effects of PGE2 on prostate cancer PC3 cells through the axis iPGE2‐iEP receptors‐EGFR‐HIF‐1α. Thus, this axis might be responsible for the growth‐stimulating effects of PGE2 on prostate epithelial cells, thereby contributing to prostate proliferative diseases associated with chronic inflammation. Since this PGT‐dependent non‐canonic intracrine mechanism of PGE2 action operates in both benign and malignant prostate epithelial cells, PGT inhibitors should be tested as a novel therapeutic modality to treat prostate proliferative disease.In this contribution we have found in immortalized, non‐malignant prostate epithelial RWPE‐1 cells that PGE2, through the axis intracellular PGE2‐iEP receptors‐EGFR‐HIF‐1α, inhibits cell adhesion and stimulates cell proliferation, migration and in vitro angiogenesis (i.e., the same effects than we have previously described in PC3 cells). Since intracellular PGE2 mediates the growth‐stimulating effects of PGE2 on prostate epithelial cells, regardless they are benign or malign ones, the pharmacological inhibition of the prostaglandin trasnporter PGT could be a novel therapeutic approach to treat prostate proliferative diseases associated with chronic inflammation.
      PubDate: 2017-11-20T07:10:30.688967-05:
      DOI: 10.1002/jcp.26220
  • NEAT1 acts as an inducer of cancer stem cell‐like phenotypes in NSCLC by
           inhibiting EGCG‐upregulated CTR1
    • Authors: Pan Jiang; Aochang Chen, Xiaoyue Wu, Ming Zhou, Ijaz ul Haq, Zahula Mariyam, Qing Feng
      Abstract: Long non‐coding RNAs (lncRNAs) play significant roles in the pathogenesis of various cancers, including lung cancer. In this study, we aimed to investigate the biological function of lncRNA nuclear enriched abundant transcript 1 (NEAT1) in cancer stem cells (CSCs). CSCs have been suggested as the main cause of tumor metastasis, tumor recurrence and chemotherapy resistance. The copper transporter 1 (CTR1) has been the focus of many recent studies because of its correlation with cisplatin (CDDP) resistance. So far, the mechanism of how NEAT1 regulates CSCs in NSCLC remains unknown. In the current study, lung cancer stem cells were enriched from the parental NSCLC cells. We observed that NEAT1 was up‐regulated while copper transporter 1 (CTR1) was down‐regulated in the enriched NSCLC cancer stem cells. Knockdown of NEAT1 was able to decrease the CSC‐like properties in NSCLC cells, while over‐expression of NEAT1 could contribute to the stemness respectively. Meanwhile, appropriate doses of EGCG restrained the stemness triggered by over‐expressing NEAT1 via inducing CTR1 expression. Wnt signal pathway and epithelial‐to‐mesenchymal transition (EMT) process were involved in NEAT1‐induced CSCs in NSCLC. These findings may suggest a novel role of NEAT1 for NSCLC treatment. This article is protected by copyright. All rights reserved
      PubDate: 2017-11-19T23:25:21.093654-05:
      DOI: 10.1002/jcp.26288
  • Molecular Mechanisms of Drug Resistance in Ovarian Cancer
    • Authors: Leyla Norouzi-Barough; Mohammad Reza Sarookhani, Mohammadreza Sharifi, Sahar Moghbelinejad, Saranaz Jangjoo, Rasoul Salehi
      Abstract: Ovarian cancer is the most lethal malignancy among the gynecological cancers, with a 5‐year survival rate, mainly due to being diagnosed at advanced stages, recurrence and resistance to the current chemotherapeutic agents. Drug resistance is a complex phenomenon and the number of known involved genes and cross‐talks between signaling pathways in this process is growing rapidly. Thus, discovering and understanding the underlying molecular mechanisms involved in chemo‐resistance are crucial for management of treatment and identifying novel and effective drug targets as well as drug discovery to improve therapeutic outcomes.In this review, the major and recently identified molecular mechanisms of drug resistance in ovarian cancer from relevant literature have been investigated.In the final section of the paper, new approaches for studying detailed mechanisms of chemo‐resistance have been briefly discussed. This article is protected by copyright. All rights reserved
      PubDate: 2017-11-19T23:20:21.425433-05:
      DOI: 10.1002/jcp.26289
  • A novel epididymal quiescence factor inhibits sperm motility by modulating
           NOS activity and intracellular NO‐cGMP pathway
    • Authors: Prasanta Ghosh; Sandipan Mukherjee, Arpita Bhoumik, Sandhya Rekha Dungdung
      Abstract: Mature and potentially motile spermatozoa stored in cauda epididymis in an inactive state for approximately 30 days; however during ejaculation they regain motility. To understand the actual molecular mechanism of the sperm quiescence during caudal stay, a proteinaceous quiescence factor (QF) has been purified from caprine epididymal plasma to apparent homogeneity. In the present study complete purification, detailed characterization as well as mechanistic pathway of QF has been described. QF was purified to 215 fold with 45% activity recovery. It is a 59 kDa monomeric protein with isoelectric point 5.8 and optimally active at pH 7.5. Circular dichroism spectroscopy and atomic force microscopy study confirm its α‐helical secondary structure and globular tertiary conformation. QF is a thermo‐stable protein as higher temperature does not alter its helical structure. N‐terminal amino acid sequencing and MALDI analysis of QF did not find 100% similarity with any available protein of the database, proved its novelty. QF at 2 μM dose inhibits sperm progressive forward motility within 10 min. This motility inhibitory activity of QF is mediated by reducing NOS enzyme activity and subsequently decreasing the intracellular NO and cGMP concentration. It does not modulate intracellular Ca++ and cAMP concentration. QF has no adverse effect on DNA integrity and morphology of spermatozoa. Motility inhibitory action of QF is reversible. Thus, the role of QF in maintaining energy saving quiescence state of mature cauda spermatozoa and its reactive nitrogen species reducing activity may lead to a new direction for storage of spermatozoa and idiopathic male infertility. This article is protected by copyright. All rights reserved
      PubDate: 2017-11-18T11:00:41.914808-05:
      DOI: 10.1002/jcp.26275
  • The roles of Wnt/β‐catenin pathway in tissue development and
           regenerative medicine
    • Authors: Maryam Majidinia; Javad Aghazadeh, Rana Jahanban-Esfahlani, Bahman Yousefi
      Abstract: Regenerative medicine is a translational field which combines tissue engineering and molecular biology to construct spare organs or help injured or defective tissues to regenerate or restore their normal functions. This is particularly important with specific organs such as heart, central nervous system, retina or limbs which possess very limited regenerative capacity. As such, regenerative medicine has received peculiar attention in the last decade. In this regard, Wnt/β‐catenin signaling pathway has been subject to intensive research, since it plays many essential roles in the regulation of the progenitor cell fate, developmental decisions, proliferation during embryonic development, and adult tissue homeostasis. In this paper, we will briefly introduce Wnt/β‐catenin signaling pathway and discuss how it integrally contributes to both stem and cancer stem cell maintenance. Finally, we summarize the current understanding of the role of Wnt/β‐catenin signaling in the development and regeneration of heart, lung, liver, bone, and cartilage. This article is protected by copyright. All rights reserved
      PubDate: 2017-11-18T10:55:34.239117-05:
      DOI: 10.1002/jcp.26265
  • Generation of insulin‐producing cells from human adipose‐derived
           mesenchymal stem cells on PVA scaffold by optimized differentiation
    • Authors: Seyed Ehsan Enderami; Masoud Soleimani, Yousef Mortazavi, Samad Nadri, Ali Salimi
      Abstract: The studies have been done on patient‐specific human adipose‐derived from mesenchymal stem cells (hADSCs) like a series of autologous growth factors and nanofibrous scaffolds (3D culture) will probably have many benefits for regenerative medicine in type 1 diabetes mellitus (TIDM) patients in the future. For this purpose, we established a polyvinyl alcohol (PVA) scaffold and a differentiation protocol by adding platelet‐rich plasma (PRP) that induces the hADSCs into insulin‐producing cells (IPCs). The Characteristics of the derived IPCs in 3D culture were compared with conventional culture (2D) groups evaluated at the mRNA and protein levels. The viability of induced pancreatic cells was 14 days. The in vitro studies showed that the treatment of hADSCs in the 3D culture resulted in differentiated cells with strong characteristics of IPCs including pancreatic‐like cells, the expression of the islet‐associated genes at the mRNA and protein levels in comparison of 2D culture group. Furthermore, the immunoassay tests showed that these differentiated cells in these two groups are functional and secreted C‐peptide and insulin in a glucose stimulation challenge. The results of our study for the first time demonstrated that the PVA nanofibrous scaffolds along with the optimized differentiation protocol with PRP can enhance the differentiation of IPCs from hADSCs. In conclusion, this study provides a new approach to the future pancreatic tissue engineering and beta cell replacement therapies for T1DM. This article is protected by copyright. All rights reserved
      PubDate: 2017-11-18T10:55:32.359812-05:
      DOI: 10.1002/jcp.26266
  • Toll like receptor signaling pathway as a potential therapeutic target in
           colorectal cancer
    • Authors: Reyhaneh Moradi-Marjaneh; Seyed Mahdi Hassanian, Hamid Fiuji, Saman Soleimanpour, Gordon A. Ferns, Amir Avan, Majid Khazaei
      Abstract: Toll like receptor (TLR) signaling is involved in activating innate and adaptive immune responses and plays a critical role in inflammation‐induced diseases such as colorectal cancer (CRC). Dysregulation of this signaling pathway can result in disturbance of epithelial layer hemostasis, chronic inflammatory, excessive repair responses and development of CRC. There is now substantial evidence for the benefit of targeting of this pathway in cancer treatment, and several agents have been approved, such as BCG (Bacillus Calmette Guérin), MPL (monophosphoryl lipid A) and imiquimod. This review summarizes the current knowledge about the different functions of TLRs on tumor cells and their application in cancer therapy with particular emphasis on recent preclinical and clinical research in treatment of CRC. This article is protected by copyright. All rights reserved
      PubDate: 2017-11-18T10:55:30.287754-05:
      DOI: 10.1002/jcp.26273
  • MicroRNA‐497 Accelerates Apoptosis While Inhibiting Proliferation,
           Migration, and Invasion through negative regulation of the MAPK/ERK
           signaling pathway via RAF‐1
    • Authors: Ling Tao; Xia Li, Na-Na Han, Qi Zhou, Zhi-Le Liu
      Abstract: The aim of this study is to explore the various modes of action miR‐497 has on human cervical cancer (CC) cell behavior. We also speculate that miR‐497 achieves its anti‐tumor role by governing RAF‐1 via MAPK/ERK signaling pathway. CC tissues with corresponding adjacent normal tissues were collected from 168 CC patients. RAF‐1‐positive cells were identified by means of immunohistochemistry in tissues. A series of inhibitors, mimics and siRNA against RAF‐1 were introduced to validate regulatory mechanisms for miR‐497 and RAF‐1. Quantitative real‐time polymerase chain reaction (qRT‐PCR) and western blot assay were employed for evaluating alternations of miR‐497, RAF‐1, and MAPK/ERK signaling pathway. HeLa cell proliferation, invasion, migration, cycle progression, and apoptosis were assessed by means of CCK‐8, wound‐healing, transwell invasion assays, and flow cytometry, respectively. The target prediction program and luciferase activity determination were used to identify miR‐497 targeting RAF‐1. We determined reduced miR‐497 expression and elevated expression of RAF‐1 in CC tissues as opposed to adjacent tissues. Transfection of miR‐497 mimics and siRNA‐RAF‐1 both decreased levels of MEK1, ERK1, and p38 phosphorylation in HeLa cells, inhibited cell proliferation, migration and invasion, induced more cells arrested in the G0/G1 phase, and promoted cell apoptosis; while miR‐497 inhibitors led to opposite results. These findings indicate miR‐497 as a tumor suppressor results from negative regulation of the MAPK/ERK signaling pathway via RAF‐1 in CC. This article is protected by copyright. All rights reserved
      PubDate: 2017-11-18T10:55:28.703401-05:
      DOI: 10.1002/jcp.26272
  • miR‐34b‐5p inhibition attenuates lung inflammation and apoptosis in an
           LPS‐induced acute lung injury mouse model by targeting progranulin
    • Authors: Wang Xie; Qingchun Lu, Kailing Wang, Jingjing Lu, Xia Gu, Dongyi Zhu, Fanglei Liu, Zhongliang Guo
      Abstract: Background: Inflammation and apoptosis play important roles in the initiation and progression of acute lung injury (ALI). Our previous study has shown that progranulin (PGRN) exerts lung protective effects during LPS‐induced ALI. Here, we have investigated the potential roles of PGRN‐targeting microRNAs (miRNAs) in regulating inflammation and apoptosis in ALI and have highlighted the important role of PGRN. Methods: LPS‐induced lung injury and the protective roles of PGRN in ALI were first confirmed. The function of miR‐34b‐5p in ALI was determined by transfection of a miR‐34b‐5p mimic or inhibitor in intro and in vivo. Results: The PGRN level gradually increased and subsequently significantly decreased, reaching its lowest value by 24 h; PGRN was still elevated compared to the control. The change was accompanied by a release of inflammatory mediators and accumulation of inflammatory cells in the lungs. Using bioinformatics analysis and RT‐PCR, we demonstrated that, among 12 putative miRNAs, the kinetics of the miR‐34b‐5p levels were closely associated with PGRN expression in the lung homogenates. The gain‐ and loss‐of‐function analysis, dual‐luciferase reporter assays and rescue experiments confirmed that PGRN was ta functional target of miR‐34b‐5p. Intravenous injection of miR‐34b‐5p antagomir in vivo significantly inhibited miR‐34b‐5p up‐regulation, reduced inflammatory cytokine release, decreased alveolar epithelial cell apoptosis, attenuated lung inflammation and improved survival by targeting PGRN during ALI. Conclusion: miR‐34b‐5p knockdown attenuates lung inflammation and apoptosis in an LPS‐induced ALI mouse model by targeting PGRN. This study shows that miR‐34b‐5p and PGRN may be potential targets for ALI treatments. This article is protected by copyright. All rights reserved
      PubDate: 2017-11-18T10:55:24.714852-05:
      DOI: 10.1002/jcp.26274
  • Tumor‐suppressive roles of ΔNp63β‐miR‐205 axis in
           epithelial‐mesenchymal transition of oral squamous cell carcinoma via
           targeting ZEB1 and ZEB2
    • Authors: Yuma Hashiguchi; Shintaro Kawano, Yuichi Goto, Kaori Yasuda, Naoki Kaneko, Taiki Sakamoto, Ryota Matsubara, Teppei Jinno, Yasuyuki Maruse, Hideaki Tanaka, Masahiko Morioka, Taichi Hattori, Shoichi Tanaka, Tamotsu Kiyoshima, Seiji Nakamura
      Abstract: We previously revealed that epithelial‐to‐mesenchymal transition (EMT) was mediated by ΔNp63β, a splicing variant of ΔNp63, in oral squamous cell carcinoma (OSCC). Recent studies have highlighted the involvement of microRNA (miRNA) in EMT of cancer cells, though the mechanism remains unclear. To identify miRNAs responsible for ΔNp63β‐mediated EMT, miRNA microarray analyses were performed by ΔNp63β‐overexpression in OSCC cells; SQUU‐B, which lacks ΔNp63 expression and displays EMT phenotypes. miRNAs microarray analyses revealed miR‐205 was the most up‐regulated following ΔNp63β‐overexpression. In OSCC cells, miR‐205 expression was positively associated with ΔNp63 and negatively with zinc‐finger E‐box binding homeobox (ZEB) 1 and ZEB2, potential targets of miR‐205. miR‐205 overexpression by miR‐205 mimic transfection into SQUU‐B cells led to decreasing ZEB1, ZEB2, and mesenchymal markers, increasing epithelial markers, and reducing cell motilities, suggesting inhibition of EMT phenotype. Interestingly, the results opposite to this phenomenon were obtained by transfection of miR‐205 inhibitor into OSCC cells, which express ΔNp63 and miR‐205. Furthermore, target protector analyses revealed direct regulation by miR‐205 of ZEB1 and ZEB2 expression. These results showed tumor‐suppressive roles of ΔNp63β and miR‐205 by inhibiting EMT thorough modulating ZEB1 and ZEB2 expression in OSCC. This article is protected by copyright. All rights reserved
      PubDate: 2017-11-18T10:55:22.589224-05:
      DOI: 10.1002/jcp.26267
  • Methylation‐associated DOK1 and DOK2 down‐regulation: potential
           biomarkers for predicting adverse prognosis in acute myeloid leukemia
    • Authors: Pin-fang He; Zi-jun Xu, Jing-dong Zhou, Xi-xi Li, Wei Zhang, De-hong Wu, Zhi-hui Zhang, Xin-Yue Lian, Xin-yu Yao, Zhao-qun Deng, Jiang Lin, Jun Qian
      Abstract: DOK‐1 and DOK‐2 (DOK1/2) are closely related members of downstream of tyrosine kinase (DOK) family genes, which are found to be frequently rearranged in several hematopoietic cancers. However, the clinical implications of DOK1/2 in acute myeloid leukemia (AML) remain largely unknown. To investigate the clinical significance, real‐time quantitative PCR (RQ‐PCR) was carried out to detect DOK1/2 expressions in 125 de novo AML patients and 28 healthy controls. Real‐time quantitative methylation‐specific PCR (RQ‐MSP) and bisulfite sequencing PCR (BSP) were applied to detect DOK1/2 methylation level and density. DOK1/2 expressions were significantly down‐regulated in AML patients. The promoters of DOK1/2 were highly hypermethylated and negatively correlated with DOK1/2 expressions in AML patients. In addition, we also confirmed that DOK1/2 expressions could be restored by DOK1/2 demethylation using 5‐aza‐2′‐deoxycytidine in leukemia cell line THP‐1. Survival analyses showed that low‐expressed DOK1/2 were associated with markedly shorter overall survival and leukemia free survival in both whole‐cohort AML and non‐M3 AML patients. Multivariate analyses further revealed that DOK1/2 were act as independent prognostic factors in AML patients. These findings indicate that decreased DOK1/2 expressions associated with their promoter hypermethylations predict adverse prognosis in AML. This article is protected by copyright. All rights reserved
      PubDate: 2017-11-18T10:50:20.853805-05:
      DOI: 10.1002/jcp.26271
  • NONO ubiquitination is mediated by FBW7 and GSK3 β via a degron lost upon
           chromosomal rearrangement in cancer
    • Authors: Luigi Alfano; Antonella Caporaso, Angela Altieri, Caterina Costa, Iris M. Forte, Carmelina A. Iannuzzi, Daniela Barone, Luca Esposito, Antonio Giordano, Francesca Pentimalli
      Abstract: NONO is an RNA‐binding protein involved in transcription, mRNA splicing, DNA repair and checkpoint activation in response to UV radiation. NONO expression has been found altered in several tumour types, including prostate, colon, breast, melanoma and in papillary renal carcinoma, in which an X chromosome inversion generates a NONO‐TFE3 fusion protein. Upon such rearrangement, NONO loses its C‐terminal domain. Through bioinformatics analysis, we identified a putative degron motif, known to be recognized by the Skp1‐Cul1‐F‐box‐protein (SCF) complex. Here, we evaluated how this domain could affect NONO protein biology. We showed that NONO interacts with the nuclear FBW7α isoform and its ubiquitination is regulated following modulation of the GSK3β kinase. Mutation of T428A/T432A within the degron impaired polyubiquitination upon FBW7α and GSK3β overexpression. Overall, our data suggest that NONO is likely subjected to proteasome‐mediated degradation and add NONO to the list of proteins targeted by FBW7, which is itself often deregulated in cancer. This article is protected by copyright. All rights reserved
      PubDate: 2017-11-18T10:25:33.329522-05:
      DOI: 10.1002/jcp.26269
  • High fat diet‐induced oxidative stress blocks hepatocyte nuclear factor
           4α and leads to hepatic steatosis in mice
    • Authors: Dongsheng Yu; Gang Chen, Minglin Pan, Jia Zhang, Wenping He, Yang Liu, Xue Nian, Liang Sheng, Bin Xu
      Abstract: Nonalcoholic fatty liver disease (NAFLD) is the most common form of chronic liver disease with manifestation of over‐accumulation of fat in liver. Increasing evidences indicate that NAFLD may be in part caused by malfunction of very low density lipoprotein (VLDL) secretion. Hepatocyte nuclear factor 4α (HNF4α), a nuclear receptor protein, plays an important role in sustain hepatic lipid homeostasis via transcriptional regulation of genes involved in secretion of VLDL, such as apolipoprotein B (ApoB). However, the exact functional change of HNF4α in NAFLD remains to be elucidated. In the present study, we found that high fat diet (HFD) induced cytoplasmic retention of HNF4α in hepatocytes, which led to down‐regulation of hepatic ApoB expression and its protein level in serum, as well as reduced secretion of VLDL. We further revealed that oxidative stress, elevated in fatty liver, was the key factor inducing the cytoplasmic retention of HNF4α in hepatocytes by activating protein kinase C (PKC)‐mediated phosphorylation in HNF4α. Thus, our findings reveal a novel mechanism underlying HFD‐induced fatty liver that oxidative stress impairs function of HNF4α on ApoB expression and VLDL secretion via PKC activation, eventually promoting fat accumulation in the liver. Therefore, oxidative stress/PKC/HNF4α pathway may be a novel target to treat diet‐induced fatty liver. This article is protected by copyright. All rights reserved
      PubDate: 2017-11-18T10:25:31.446552-05:
      DOI: 10.1002/jcp.26270
  • Regulation of Calcification in Human Aortic Smooth Muscle Cells Infected
           with High‐glucose‐treated Porphyromonas Gingivalis
    • Authors: Te-Chuan Chen; Chien-Tsong Lin, Shao-Ju Chien, Shun-Fu Chang, Cheng-Nan Chen
      Abstract: Porphyromonas (P.) gingivalis infection leading to the periodontitis has been associated with the development of systemic diseases, including cardiovascular diseases and diabetes. However, the effect of a high concentration of glucose (HG) on the invasion efficiency of P. gingivalis and the consequent modulation of pathogenesis in vascular cells, especially in the vascular smooth muscle cells (VSMCs), remains unclear. Hence, the aim of this study was to investigate whether treating P. gingivalis with HG could change its invasion capability and result in VSMC calcification and the underlying mechanism. Human aortic SMCs (HASMCs) and P. gingivalis strain CCUG25226 were used in this study. We found that HGPg infection of HASMCs could initiate the HASMC calcification by stimulating the autocrine regulation of bone morphogenetic protein (BMP) 4 in HASMCs. The upregulation of BMP4 expression in HASMCs was mediated by toll‐like receptor 4 and ERK1/2‐p38 signaling after P. gingivalis infection. Moreover, the autocrine action of BMP4 in HGPg infection‐initiated HASMC calcification upregulated BMP4‐specific downstream smad1/5/8‐runx2 signaling to increase the expressions of bone‐related matrix proteins, i.e. osteopontin, osteocalcin, and alkaline phosphatase. This study elucidates the detailed mechanism of HGPg infection‐initiated calcification of HASMCs and indicates a possible therapeutic role of BMP4 in P. gingivalis infection‐associated vascular calcification. This article is protected by copyright. All rights reserved
      PubDate: 2017-11-18T10:25:19.610277-05:
      DOI: 10.1002/jcp.26268
  • LncRNA‐TCONS_00034812 in cell proliferation and apoptosis of pulmonary
           artery smooth muscle cells and its mechanism
    • Authors: Yun Liu; Zengxian Sun, Jinquan Zhu, Bingxin Xiao, Jie Dong, Xiaomin Li
      Abstract: Long noncoding RNAs (lncRNAs) have been discovered to be playing important role in various biological processes. However, the contribution of lncRNAs to pulmonary artery hypertension (PAH) remains largely unknown. Pulmonary vascular remodeling is an important pathological feature of PAH, leading to increased vascular resistance and reduced compliance. Here, we investigated the biological role of lncRNAs in PAH. Differences in the lncRNAs and mRNAs between hypoxia PAH rats and normoxia rats were screened using microarray analysis. The results showed that 36 lncRNAs and 519 mRNAs were upregulated in the pulmonary arteries (PAs) of hypoxia PAH rats, whereas 111 lncRNAs and 246 mRNAs were downregulated. Expressions of the screened lncRNAs, including TCONS_00034812, were validated by real‐time PCR. We revealed that the expression of TCONS_00034812 was significantly downregulated in PAs of PAH rats and hypoxia pulmonary artery smooth muscle cells (PASMCs). TCONS_00034812 knockdown promoted proliferation and inhibited apoptosis of PASMCs in vitro. Moreover, TCONS_00034812 regulated PASMCs function in vitro. We found that TCONS_00034812 increased the expression of transcription factors Stox1. TCONS_00034812 and Stox1 knockdown mediated PASMCs function through MAPK signaling. Our findings imply lncRNA as a critical regulator in PAH and demonstrate the potential of gene therapy and drug development for treating PAH. The present study reveals a novel mechano responsive lncRNA‐ TCONS_00034812, which modulates PASMCs proliferation and apoptosis, and participates in vascular remodelling during PAH. This article is protected by copyright. All rights reserved
      PubDate: 2017-11-18T09:10:33.945272-05:
      DOI: 10.1002/jcp.26279
    • Authors: Maria Guitart; Josep Lloreta, Laura Mañas-Garcia, Esther Barreiro
      Abstract: Reduced muscle activity leads to muscle atrophy and function loss in patients and animal models. Satellite cells (SCs) are postnatal muscle stem cells that play a pivotal role in skeletal muscle regeneration following injury. The regenerative potential, satellite cell numbers, and markers during recovery following immobilization of the hindlimb for seven days were explored. In mice exposed to seven days of hindlimb immobilization, in those exposed to recovery (seven days, splint removal), and in contralateral control muscles, muscle precursor cells were isolated from all hindlimb muscles (fluorescence‐activated cell sorting, FACS), and SCs and muscle regeneration were identified using immunofluorescence (gastrocnemius and soleus) and electron microscopy (EM, gastrocnemius). Expression of ki67, pax7, myoD, and myogenin was quantified (RT‐PCR) from SC FACS yields. Body and grip strength were determined. Following seven‐day hindlimb immobilization, a decline in SCs (FACS, immunofluorescence) was observed together with an upregulation of SC activation markers and signs of muscle regeneration including fusion to existing myofibers (EM). Recovery following hindlimb immobilization was characterized by a program of muscle regeneration events. Hindlimb immobilization induced a decline in SCs together with an upregulation of markers of SC activation, suggesting that fusion to existing myofibers takes place during unloading. Muscle recovery induced a significant rise in muscle precursor cells and regeneration events along with reduced SC activation expression markers and a concomitant rise in terminal muscle differentiation expression. These are novel findings of potential applicability for the treatment of disuse muscle atrophy, which is commonly associated with severe chronic and acute conditions. This article is protected by copyright. All rights reserved
      PubDate: 2017-11-18T09:10:26.881168-05:
      DOI: 10.1002/jcp.26282
  • Development of an In Vivo Mouse Model of Discogenic Low Back Pain
    • Authors: Changgui Shi; Vaskar Das, Xin Li, Ranjan Kc, Sujun Qiu, InSug O-Sullivan, Richard L Ripper, Jeffrey S Kroin, Fackson Mwale, Atiyayein A. Wallace, Bingqian Zhu, Lan Zhao, Andre J. van Wijnen, Mingliang Ji, Jun Lu, Gina Votta-Velis, Wen Yuan, Hee-Jeong Im
      Abstract: Discogenic Low back pain (DLBP) is extremely common and costly. Effective treatments are lacking due to DLBP's unknown pathogenesis. Currently, there are no in vivo mouse models of DLBP, which restricts research in this field. The aim of this study was to establish a reliable DLBP model in mouse that captures the pathological changes in the disc and allows longitudinal pain testing. The model was generated by puncturing the mouse lumbar discs (L4/5, L5/6, and L6/S1) and removing the nucleus pulposus using a microscalpel under the microscope. Histology, molecular pathways, and pain‐related behaviors were examined. Over 12 weeks post‐surgery, animals displayed the mechanical, heat, and cold hyperalgesia along with decreased burrowing and rearing. Histology showed progressive disc degeneration with loss of disc height, nucleus pulposus reduction, proteoglycan depletion, and annular fibrotic disorganization. Immunohistochemistry revealed a substantial increase in inflammatory mediators at 2 and 4 weeks. Nerve growth factor was upregulated from 2 weeks to the end of the experiment. Nerve fiber ingrowth was induced in the injured discs after 4 weeks. Disc‐puncture also produced an upregulation of neuropeptides in dorsal root ganglia neurons and an activation of glial cells in the spinal cord dorsal horn. These findings indicate that the cellular and structural changes in discs, as well as peripheral and central nervous system plasticity, paralleled persistent and robust behavioral pain responses. Therefore, this mouse DLBP model could be used to investigate mechanisms underlying discogenic pain, thereby facilitating effective drug screening and development of treatments for DLBP. This article is protected by copyright. All rights reserved
      PubDate: 2017-11-18T09:05:28.100345-05:
      DOI: 10.1002/jcp.26280
  • The current status and perspectives regarding the clinical implication of
           intracellular calcium in breast cancer
    • Authors: Amir Tajbakhsh; Alireza Pasdar, Mehdi Rezaee, Mostafa Fazeli, Saman Soleimanpour, Seyed Mahdi Hassanian, Zahra FarshchiyanYazdi, Tayebe Younesi Rad, Gordon A. Ferns, Amir Avan
      Abstract: Calcium ions (Ca2+) act as second messengers in intracellular signaling. Ca2+ pumps, channels, sensors, and calcium binding proteins, regulate the concentrations of intracellular Ca2+ as a key regulator of important cellular processes such as gene expression, proliferation, differentiation, DNA repair, apoptosis, metastasis, and hormone secretion. Intracellular Ca2+ also influences the functions of several organelles, that include: the endoplasmic reticulum, mitochondria, the Golgi, and cell membrane both in normal and breast cancer cells. In breast cancer, the disruption of intracellular: Ca2+ homeostasis may cause tumor progression by affecting key factors/pathways including phospholipase C (PLC), inositol 1,4,5‐trisphosphate (IP3), calmodulin (CaM), nuclear factor of activated T‐cells (NFAT), calpain, calmodulin‐dependent protein kinase II (CaMKII), mitogen‐activated protein kinase (MAPK), epithelial‐mesenchymal transition (EMT), vascular endothelial growth factor (VEGF), poly (ADP‐Ribose) polymerase‐1 (PARP1), estrogen, and estrogen receptor. Because the foregoing molecules play crucial roles in breast cancer, the factors/pathways influencing intracellular Ca2+ concentrations are putative targets for cancer treatment, using drugs such as Mephebrindole, Tilapia piscidin 4, Nifetepimine, Paricalcitol, and Prednisolone. We have explored the factors/pathways which are related to breast cancer and Ca2+ homeostasis and signaling in this review, and also discussed their potential as biomarkers for breast cancer staging, prognosis, and therapy.
      PubDate: 2017-11-18T09:05:25.177735-05:
      DOI: 10.1002/jcp.26277
  • Physiological role of urothelial cancer‐associated 1 long noncoding RNA
           in human skeletogenic cell differentiation
    • Authors: Takanori Ishikawa; Takashi Nishida, Mitsuaki Ono, Takeshi Takarada, Nguyen Ha, Shinnosuke Kurihara, Takayuki Furumatsu, Yurika Murase, Masaharu Takigawa, Toshitaka Oohashi, Hiroshi Kamioka, Satoshi Kubota
      Abstract: A vast number of long‐noncoding RNAs (lncRNA) are found expressed in human cells, which RNAs have been developed along with human evolution. However, the physiological functions of these lncRNAs remain mostly unknown. In the present study, we for the first time uncovered the fact that one of such lncRNAs plays a significant role in the differentiation of chondrocytes and, possibly, of osteoblasts differentiated from mesenchymal stem cells, which cells eventually construct the human skeleton. The urothelial cancer‐associated 1 (UCA1) lncRNA is known to be associated with several human malignancies. Firstly, we confirmed that UCA1 was expressed in normal human chondrocytes, as well as in a human chondrocytic cell line; whereas it was not detected in human bone marrow mesenchymal stem cells (hBMSCs). Of note, although UCA1 expression was undetectable in hBMSCs, it was markedly induced along with the differentiation towards chondrocytes, suggesting its critical role in chondrogenesis. Consistent with this finding, silencing of the UCA1 gene significantly repressed the expression of chondrogenic genes in human chondrocytic cells. UCA1 gene silencing and hyper‐expression also had a significant impact on the osteoblastic phenotype in a human cell line. Finally, forced expression of UCA1 in a murine chondrocyte precursor, which did not possess a UCA1 gene, overdrove its differentiation into chondrocytes. These results indicate a physiological and important role of this lncRNA in the skeletal development of humans, who require more sustained endochondral ossification and osteogenesis than do smaller vertebrates. This article is protected by copyright. All rights reserved
      PubDate: 2017-11-18T08:55:30.742314-05:
      DOI: 10.1002/jcp.26285
  • Epithelial Cell Adhesion Molecule Fragments and Signaling in Primary Human
           Liver Cells
    • Authors: Jörg C. Gerlach; Hubert G. Foka, Robert L. Thompson, Bruno Gridelli, Eva Schmelzer
      Abstract: Epithelial Cell Adhesion Molecule (EpCAM), or CD326, is a trans‐membrane glycoprotein expressed by multiple normal epithelia as well as carcinoma. Human hepatic stem cells and bile duct epithelium of the liver are EpCAM positive. In tumor cell lines, its intracellular domain can be released after cleavage of the extracellular domain. Within the cell nucleus it induces cell proliferation, but cleavage depends on cell contact. Fragments of various lengths have been described in tumor cells. Despite of its described important role in proliferation in tumor cells, there is not much known about the expression and role of EpCAM fragments in primary human liver cells. Here, we demonstrate that EpCAM protein fragments and function are considerable different between tumor cells, normal fetal and adult liver cells. Contrary to previously reported findings in tumor cells, gene knockdown or treatment with an inhibitor of the cleavage enzyme ADAM17 (TACE) rather increased cell numbers in primary human fetal liver‐derived EpCAM‐positive cells. EpCAM fragment sizes were not affected by treatment with inhibitor. Knockdown of EPCAM gene expression by siRNA in sorted cells did not significantly affect proliferation‐associated genes or cell numbers. The intracellular domain could not be detected within cell nuclei of fetal and adult liver cells. In conclusion, signaling through the intracellular domain of EpCAM appears to be a mechanism that induces proliferation specifically in tumorigenic cells but not in normal primary EpCAM‐positive liver cells. This article is protected by copyright. All rights reserved
      PubDate: 2017-11-18T08:55:25.989672-05:
      DOI: 10.1002/jcp.26286
  • Inflammasome: its role in traumatic brain and spinal cord injury
    • Authors: Keywan Mortezaee; Neda Khanlarkhani, Cordian Beyer, Adib Zendedel
      Abstract: Traumatic brain injury (TBI) and spinal cord injury (SCI) are pathological events that lead to neuropathological conditions which have in consequence the initiation of pro‐inflammatory cytokine production. Neuroinflammation plays a key role in the secondary phase of both TBI and SCI after initial cell death. Activation of cytoplasmic inflammasome complexes is regarded as the essential step of neuroinflammation and a key trigger for neuronal death called pyroptosis. Inflammasome complexes are involved in activation of caspase‐1 which catalyzes the cleavage of pro‐interleukins into their active forms (including interleukin‐18 (IL‐18) and IL‐1β). The focus of this article is to discuss the time‐course and regulation of inflammasome assembly and activation during TBI and SCI and their targeting in designing therapeutic approaches. We particularly focus on the inflammasomes NLRP1 and NLRP3 which play a pivotal function during TBI and SCI in the central nervous system (CNS). This article is protected by copyright. All rights reserved
      PubDate: 2017-11-18T08:55:23.504848-05:
      DOI: 10.1002/jcp.26287
  • Inhibition of miR‐200b/miR‐429 Contributes to Neuropathic Pain
           development Through Targeting Zinc finger E box binding protein‐1
    • Authors: Xue-Tao Yan; Ying Zhao, Xiao-Li Cheng, Xiang-Hu He, Yu Wang, Wen-Zhong Zheng, Hu Chen, Yan-Lin Wang
      Abstract: Many studies have reported that microRNAs participate in neuropathic pain development. Previously, miR‐200b and miR‐429 are reported to be involved in various diseases. In our current study, we focused on their roles in neuropathic pain and we found that miR‐200b and miR‐429 were significantly decreased in chronic constriction injury (CCI) rat spinal cords and isolated microglials. miR‐200b and miR‐429 overexpression were able to relieve neuropathic pain through modulating PWT and PWL in CCI rats. Meanwhile, we observed that both miR‐200b and miR‐429 upregulation could repress neuroinflammation via inhibiting inflammatory cytokines such as IL‐6, IL‐1β and TNF‐α in CCI rats. By carry out bioinformatics technology, Zinc finger E box binding protein‐1 (ZEB1) was predicted as target of miR‐200b and miR‐429 and dual‐luciferase reporter assays confirmed the correlation between them. ZEB1 has been reported to regulate a lot of diseases. Here, we found that ZEB1 was greatly increased in CCI rats and miR‐200b and miR‐429 overexpression markedly suppressed ZEB1 mRNA expression in rat microglial cells. In addition, knockdown of ZEB1 can reduce neuropathic pain development and co‐transfection of LV‐anti‐miR‐200b/miR‐429 reversed this phenomenon in vivo. Taken these together, our results suggested that miR‐200b/miR‐429 can serve as an important regulator of neuropathic pain development by targeting ZEB1. This article is protected by copyright. All rights reserved
      PubDate: 2017-11-18T08:50:27.35704-05:0
      DOI: 10.1002/jcp.26284
  • MicroRNAs as Predictive Biomarkers for Myocardial Injury in Aged Mice
           Following Myocardial Infarction
    • Authors: Natia Qipshidze Kelm; Kellianne M. Piell, Eugenia Wang, Marsha P. Cole
      Abstract: The occurrence of myocardial infarction (MI) increases appreciably with age. In the Framingham Heart Study, the incidence of MI more than doubles for men and increases more than 5‐fold in women (ages 55 to 64 years compared to 85 to 94 years). MicroRNAs (miRNAs) quantitatively regulate their target's expression post‐transcriptionally by either silencing action through binding at the 3'UTR domains or degrading the messages at their coding regions. In either case, these regulations affect the cardiac transcriptional output and cardiac function. Among the known cardiac associated miRNA, miRNA‐1, miRNA‐133a, and miRNA‐34a have been shown to induce adverse structural remodeling to impair cardiac contractile function. In the present study, an in vivo model of MI in young (3 month) and old (22 month) mice is used to investigate the possible role whereby these three miRNAs exert negative effects on heart function following MI. Herein we demonstrate that in older mouse heart, all three microRNAs show increased levels of expression, while miRNA‐1 shows a further increase in old mouse heart following MI, which corresponds to left ventricular (LV) wall thinning. These structural changes in cardiac tissue may causes downstream LV dilation and subsequent LV dysfunction. Results presented here suggest that significantly elevated levels of miRNA‐1 in post‐MI old heart could be predictive of cardiac injury in older mice as the high risk biomarker for MI in older individuals. This article is protected by copyright. All rights reserved
      PubDate: 2017-11-18T08:50:25.014723-05:
      DOI: 10.1002/jcp.26283
  • FSCN1 predicts survival and is regulated by a PI3K‐dependent
           mechanism in renal cell carcinoma
    • Authors: Mengping Zhang; Zhijian Zhao, Xiaolu Duan, Ping Chen, Zhenwei Peng, Huijuan Qiu
      Abstract: While overexpression of FSCN1 is reported in several cancers, the prognostic significance of FSCN1 in renal cell carcinoma (RCC) and the molecular mechanisms involved remain largely unclear. We retrospectively enrolled 194 patients with non‐metastatic clear‐cell RCC undergoing nephrectomy in our center between 2008 and 2011. FSCN1 expression was assessed by immunohistochemical staining and its association with clinicopathologic features and survival were evaluated. Functional effects of a modulated FSCN1 expression were analyzed with regard to invasion in RCC cell lines and metastasis in vivo. Here, we reported that FSCN1 was up‐regulated in RCC tissues compared to non‐tumor tissues, and associated with poor overall survival and recurrence‐free survival. Its expression was not associated with age, tumor size, and clinical TNM stage. The incorporation of FSCN1 into the T stage and histologic grade would help to refine individual risk stratification. Preclinical studies using multiple RCC cells and orthotopic xenografts mice model indicated that FSCN1 could promote RCC cell invasion in vitro, and metastasis in vivo. Mechanistically, overexpression of FSCN1 led to an up‐regulation of MMP9 and N‐Cadherin. Notably, treating RCC cells with PI3K/AKT inhibitors or knockdown GSK‐3β decreased the expression of FSCN1, and then attenuated RCC invasion. Together, our results demonstrate that FSCN as an oncogene is a potential novel prognostic biomarker for RCC patients after nephrectomy, and can promote RCC metastasis. This article is protected by copyright. All rights reserved
      PubDate: 2017-11-17T01:55:33.873092-05:
      DOI: 10.1002/jcp.26264
  • The MIP‐1α autocrine loop contributes to decreased sensitivity
           to anticancer drugs
    • Authors: Masanobu Tsubaki; Tomoya Takeda, Yoshika Tomonari, Kenji Mashimo, Yu-ichi Koumoto, Sachi Hoshida, Tatsuki Itoh, Motohiro Imano, Takao Satou, Katsuhiko Sakaguchi, Shozo Nishida
      Abstract: Several autocrine soluble factors, including macrophage inflammatory protein‐1α (MIP‐1α), tumor necrosis factor‐α, and hepatocyte growth factor, promote cell survival and growth in multiple myeloma (MM) cells. We hypothesized that inhibition of the MIP‐1α autocrine loop may enhance the cytotoxic effect of anticancer drugs in MM cell lines. In the present study, an MIP‐1α neutralizing antibody suppressed cell proliferation and enhanced the cytotoxic effect of melphalan or bortezomib on MM cells. In addition, melphalan resistance cells (RPMI8226/L‐PAM and HS‐sultan/L‐PAM cells) secreted MIP‐1α and neutralizing antibody of MIP‐1α partially overcame melphalan resistance. Moreover, combination treatment with MIP‐1α neutralizing antibody and melphalan or bortezomib inhibited extracellular signal regulated kinase 1/2 (ERK1/2), Akt, and mammalian target of rapamycin (mTOR) activation, Bcl‐2, Bcl‐xL, and Survivin expression, and upregulated the expression of Bim and cleaved Poly (ADP‐ribose) polymerase (PARP). Treatment of IM9 cells with MIP‐1α siRNA suppressed the activation of ERK1/2, Akt, and mTOR, and enhanced the cytotoxic effect of melphalan and bortezomib. These results indicate that MIP‐1α neutralizing antibodies or MIP‐1α siRNA enhance the cytotoxic effect of melphalan and bortezomib by suppressing the chemokine receptor/ERK and chemokine receptor/Akt/mTOR pathways. The inhibition of MIP‐1α may thus provide a new therapeutic approach to control tumor progression and bone destruction in patients with MM. This article is protected by copyright. All rights reserved
      PubDate: 2017-11-16T04:31:02.367289-05:
      DOI: 10.1002/jcp.26245
  • Synergistic effect of HIF‐1α and FoxO3a trigger cardiomyocyte apoptosis
           under hyperglycemic ischemia condition
    • Authors: Ya-Fang Chen; Sudhir Pandey, Cecilia Hsuan Day, Yu-Feng Chen, Ai-Zhi Jiang, Tsung-Jung Ho, Ray-Jade Chen, Vijaya V. Padma, Wei-Wen Kuo, Chih-Yang Huang
      Abstract: Cardiomyocyte death is an important pathogenic feature of ischemia and heart failure. Through this study, we showed the synergistic role of HIF‐1α and FoxO3a in cardiomyocyte apoptosis subjected to hypoxia plus elevated glucose levels. Using gene specific small interfering RNAs (siRNA), semi‐quantitative reverse transcriptase polymerase chain reaction (RT‐PCR), western blot, immunofluorescence, nuclear and cytosolic localization and TUNEL assay techniques, we determined that combined function of HIF‐1α and FoxO3a under high glucose plus hypoxia condition lead to enhanced expression of BNIP3 inducing cardiomyocyte death. Our results highlighted the importance of the synergistic role of HIF‐1α and FoxO3a in cardiomyocyte death which may add insight into therapeutic approaches to pathophysiology associated with ischemic diabetic cardiomyopathies. This article is protected by copyright. All rights reserved
      PubDate: 2017-11-16T04:30:36.700022-05:
      DOI: 10.1002/jcp.26235
  • Characterization and assessment of potential microRNAs involved in
           phosphate-induced aortic calcification
    • Authors: Maya Fakhry; Najwa Skafi, Mohammad Fayyad-Kazan, Firas Kobeissy, Eva Hamade, Saida Mebarek, Aida Habib, Nada Borghol, Asad Zeidan, David Magne, Hussein Fayyad-Kazan, Bassam Badran
      Abstract: Medial artery calcification, a hallmark of type 2 diabetes mellitus and chronic kidney disease (CKD), is known as an independent risk factor for cardiovascular mortality and morbidity. Hyperphosphatemia associated with CKD is a strong stimulator of vascular calcification but the molecular mechanisms regulating this process remain not fully understood. We showed that calcification was induced after exposing Sprague-Dawley rat aortic explants to high inorganic phosphate level (Pi, 6 mM) as examined by Alizarin red and Von Kossa staining. This calcification was associated with high Tissue-Nonspecific Alkaline Phosphatase (TNAP) activity, vascular smooth muscle cells de-differentiation, manifested by downregulation of smooth muscle 22 alpha (SM22α) protein expression which was assessed by immunoblot analysis, immunofluorescence, and trans-differentiation into osteo-chondrocyte-like cells revealed by upregulation of Runt related transcription factor 2 (Runx2), TNAP, osteocalcin, and osteopontin mRNA levels which were determined by quantitative real-time PCR. To unravel the possible mechanism(s) involved in this process, microRNA (miR) expression profile, which was assessed using TLDA technique and thereafter confirmed by individual qRT-PCR, revealed differential expression 10 miRs, five at day 3 and 5 at day 6 post Pi treatment versus control untreated aortas. At day 3, miR-200c, -155, 322 were upregulated and miR-708 and 331 were downregulated. After 6 days of treatment, miR-328, -546, -301a were upregulated whilst miR-409 and miR-542 were downregulated. Our results indicate that high Pi levels trigger aortic calcification and modulation of certain miRs. These observations suggest that mechanisms regulating aortic calcification might involve miRs, which warrant further investigations in future studies. This article is protected by copyright. All rights reserved
      PubDate: 2017-11-16T04:26:42.533014-05:
      DOI: 10.1002/jcp.26121
  • Horizontal transfer of miR‐23a from hypoxic tumor cell colonies can
           induce angiogenesis
    • Authors: T. V. Sruthi; Lincy Edatt, Grace R. Raji, Haritha Kunhiraman, Sharath S. Shankar, Vandana Shankar, Vishnu Ramachandran, Aswini Poyyakkara, Sameer V. B. Kumar
      Abstract: Neo vessel formation by angiogenesis is an important event during many pathological conditions including cancer, where it is indispensable for tumor growth and survival. Although, various pro‐angiogenic cytokines and soluble factors, secreted by tumor cells, have been reported to promote angiogenesis, recent studies have shown regulatory role of exosomes, secreted by tumor cells in the process of angiogenesis. These exosomes are capable of carrying nucleic acids, proteins etc. as their cargo. Under the light of these facts and considering the presence of miRNAs, the non‐coding RNAs capable of regulating target gene expression, as one of the major cargos in the exosomes, we investigated, whether exosomes derived from normoxic and hypoxic tumor cell colonies exhibit difference in levels of miR‐23∼27∼24 cluster members and if so, to check the significance of their horizontall transfer on the process of angiogenesis. Results of our study showed that exosomes secreted by hypoxic tumor cell colonies possess significantly higher levels of miR23a and can induce angiogenesis.. Further we have shown that exosomes secreted by cells that ectopically over express miR23a is capable of inducing angiogenesis in different angiogenic model systems such as CAM, in ovo Xenograft and HUVEC models systems. Further, mechanistic analysis revealed that miR23a driven regulation of angiogenesis is brought about by down regulation of SIRT1 in the recipient cells. Collectively, the results presented here suggest that exosomal transfer of miR23a from tumor cell colonies can induce the process of angiogenesis by targeting SIRT1 in the recipient endothelial cells. This article is protected by copyright. All rights reserved
      PubDate: 2017-11-16T04:10:42.368935-05:
      DOI: 10.1002/jcp.26202
  • Estrogen regulates stemness and senescence of bone marrow stromal cells to
           prevent osteoporosis via ERβ‐SATB2 pathway
    • Authors: Geng Wu; Rongyao Xu, Ping Zhang, Tao Xiao, Yu Fu, Yuchao Zhang, Yifei Du, Jinhai Ye, Jie Cheng, Hongbing Jiang
      Abstract: Decline of pluripotency in bone marrow stromal cells (BMSCs) associated with estrogen deficiency leads to a bone formation defect in osteoporosis. Special AT‐rich sequence binding protein 2 (SATB2) is crucial for maintaining stemness and osteogenic differentiation of BMSCs. However, whether SATB2 is involved in estrogen‐deficiency associated‐osteoporosis is largely unknown. In this study, we found that estrogen mediated pluripotency and senescence of BMSCs, primarily through estrogen receptor beta (ERβ). BMSCs from the OVX rats displayed increased senescence and weaker SATB2 expression, stemness, and osteogenic differentiation, while estrogen could rescue these phenotypes. Inhibition of ERβ or ERα confirmed that SATB2 was associated with ERβ in estrogen‐mediated pluripotency and senescence of BMSCs. Furthermore, estrogen mediated the upregulation of SATB2 through the induction of ERβ binding to estrogen response elements (ERE) located at ‐488 of the SATB2 gene. SATB2 overexpression alleviated senescence and enhanced stemness and osteogenic differentiation of OVX‐BMSCs. SATB2‐modified BMSCs transplantation could prevent trabecular bone loss in an ovariectomized rat model. Collectively, our study revealed the role of SATB2 in stemness, senescence and osteogenesis of OVX‐BMSCs. Collectively, these results indicate that estrogen prevents osteoporosis by promoting stemness and osteogenesis and inhibiting senescence of BMSCs through an ERβ‐SATB2 pathway. Therefore, SATB2 is a novel anti‐osteoporosis target gene. This article is protected by copyright. All rights reserved
      PubDate: 2017-11-16T04:06:18.574102-05:
      DOI: 10.1002/jcp.26233
  • LncRNA GHET1 predicts poor prognosis in hepatocellular carcinoma and
           promotes cell proliferation by silencing KLF2
    • Authors: Li Jin; Yangke He, Shijia Tang, Shan Huang
      Abstract: Background: Hepatocellular carcinoma (HCC) has been identified as one of the leading causes of cancer‐related death worldwide. Recently, long non‐coding RNAs (lncRNAs) attract much attention of researchers, and they are demonstrated to be dysregulated in a variety of cancers, including HCC. LncRNA gastric carcinoma high expressed transcript 1 lncRNA GHET1 is found to be dysregulated in gastric cancer (GC). However, its clinical value and potential biological function in HCC remains unclear.Methods and results: In this study, the expression level of GHET1 was analyzed in 72 HCC tissues and matched normal tissues by using Quantitative RT‐PCR (qRT‐PCR). GHET1 expression was significantly up‐regulated in HCC tissues and the higher level of GHET1 was related to vascular invasion, cirrhosis, tumor size, edmindson grade and poor prognosis. Moreover, knockdown of GHET1 inhibited cell proliferation of HCC, and also caused cell cycle arrest and induced apoptosis in HCC cell lines. We also found that GHET1 could epigenetically repress transcription of Kruppel‐like factor 2 (KLF2) in HCC cells by recruiting PRC2 into KLF2 promoter region.Conclusion: Our results inidcated that lncRNA GHET1, as a growth regulator, might serve as a novel prognostic biomarker and therapy target for HCC. This article is protected by copyright. All rights reserved
      PubDate: 2017-11-15T06:55:50.050863-05:
      DOI: 10.1002/jcp.26257
  • Elevated potassium outward currents in hyperoxia treated atrial
    • Authors: Z Vysotskaya; B Chidipi, J.L Rodgers, X Tang, E Samal, N Kolliputi, S Mohapatra, E.S Bennett, S.K Panguluri
      Abstract: Supplementation of 100% oxygen is a very common intervention in intensive care units (ICU) and critical care centers for patients with dysfunctional lung and lung disorders. Although there is advantage in delivering sufficient levels of oxygen, hyperoxia is reported to be directly associated with increasing in‐hospital deaths. Our previous studies reported ventricular and electrical remodeling in hyperoxia treated mouse hearts, and in this article, for the first time, we are investigating the effects of hyperoxia on atrial electrophysiology using whole‐cell patch‐clamp electrophysiology experiments along with assessment of Kv1.5, Kv4.2 and KChIP2 transcripts and protein profiles using real‐time quantitative RT‐PCR and Western blotting. Our data showed that induction of hyperoxia for 3 days in mice showed larger outward potassium currents with shorter action potential durations (APD). This increase in current densities is due to significant increase in ultrarapid delayed rectifier outward K+ currents (IKur) and rapidly activating, rapidly inactivating transient outward K+ current (Ito) densities. We also observed a significant increase in both transcripts and protein levels of Kv1.5 and KChIP2 in hyperoxia treated atrial cardiomyocytes, whereas no significant change was observed in Kv4.2 transcripts or protein. The data presented here further support our previous findings that hyperoxia induces not only ventricular remodeling, but also atrial electrical remodeling. This article is protected by copyright. All rights reserved
      PubDate: 2017-11-15T06:55:25.9111-05:00
      DOI: 10.1002/jcp.26263
  • Acrosomal alkalization triggers Ca2+ release and acrosome reaction in
           mammalian spermatozoa
    • Authors: Julio C. Chávez; José L. Vega-Beltrán, Omar José, Paulina Torres, Takuya Nishigaki, Claudia L. Treviño, Alberto Darszon
      Abstract: The sperm acrosome reaction (AR), an essential event for mammalian fertilization, involves Ca2+ permeability changes leading to exocytosis of the acrosomal vesicle. The acrosome, an intracellular Ca2+ store whose luminal pH is acidic, contains hydrolytic enzymes. It is known that acrosomal pH (pHacr) increases during capacitation and this correlates with spontaneous AR. Some AR inducers increase intracellular Ca2+ concentration ([Ca2+]i) through Ca2+ release from internal stores, mainly the acrosome. Catsper, a sperm specific Ca2+ channel, has been suggested to participate in the AR. Curiously, Mibefradil and NNC55‐0396, two CatSper blockers, themselves elevate [Ca2+]i by unknown mechanisms. Here we show that these compounds, as other weak bases, can elevate pHacr, trigger Ca2+ release from the acrosome and induce the AR in both mouse and human sperm. To our surprise, μM concentrations of NNC55‐0396 induced AR even in nominally Ca2+ free media. Our findings suggest that alkalization of the acrosome is critical step for Ca2+ release from the acrosome that leads to the acrosome reaction. This article is protected by copyright. All rights reserved
      PubDate: 2017-11-14T08:25:50.621619-05:
      DOI: 10.1002/jcp.26262
  • Tetrahydrocurcumin Ameliorates Homocysteine Mediated Mitochondrial
           Remodeling in Brain Endothelial Cells
    • Authors: Jonathan C. Vacek; Jyotirmaya Behera, Akash K. George, Pradip K. Kamat, Anuradha Kalani, Neetu Tyagi
      Abstract: Homocysteine (Hcy) causes endothelial dysfunction by inducing oxidative stress in most neurodegenerative disorders. This dysfunction is highly correlated with mitochondrial dynamics such as fusion and fission. However, there are no strategies to prevent Hcy induced mitochondrial remodeling. Tetrahydrocurcumin (THC) is an anti-inflammatory and anti-oxidant compound. We hypothesized that THC may ameliorates Hcy induced mitochondria remodeling in mouse brain endothelial cells (bEnd3) cells. bEnd3 cells were exposed to Hcy treatment in the presence or absence of THC. Cell viability and autophagic cell death were measured with MTT and MDC staining assay. Reactive oxygen species (ROS) production was determined using DCFH-DA staining by confocal microscopy. Autophagy flux was assessed using a conventional GFP-microtubule-associated protein 1 light chain 3 (LC3) dot assay. Interaction of phagophore marker LC-3 with mitochondrial receptor NIX was observed by confocal imaging. Mitochondrial fusion and fission were evaluated by western blot and RT-PCR. Our results demonstrated that Hcy resulted in cell toxicity in a dose-dependent manner and supplementation of THC prevented the detrimental effects of Hcy on cell survival. Furthermore, Hcy also upregulated of fission marker (DRP-1), fusion markers (Mfn2) and autophagy marker (LC-3). Finally, we observed that Hcy activated mitochondrial specific phagophore marker (LC-3) was co-localized with the mitochondrial receptor NIX, as viewed by confocal microscopy. Pretreatment of bEnd3 with THC (15µM) ameliorated Hcy induced oxidative damage, mitochondrial fission/fusion, and mitophagy. Our studies strongly suggest that THC has beneficial effects on mitochondrial remodeling and could be developed as a potential therapeutic agent against hyperhomocysteinemia (HHcy) induced mitochondrial dysfunction. This article is protected by copyright. All rights reserved
      PubDate: 2017-11-14T00:40:29.776906-05:
      DOI: 10.1002/jcp.26145
  • Chrysin Attenuates Progression of Ovarian Cancer Cells by Regulating
           Signaling Cascades and Mitochondrial Dysfunction
    • Authors: Whasun Lim; Soomin Ryu, Fuller W. Bazer, Sung-Man Kim, Gwonhwa Song
      Abstract: Chrysin is mainly found in passion flowers, honey, and propolis acts as a potential therapeutic and preventive agent to inhibit proliferation and invasion of various human cancer cells. Although chrysin has anti-carcinogenic effects in several cancers, little is known about its functional roles in ovarian cancer which shows poor prognosis and chemoresistance to traditional therapeutic agents. In the present study, we investigated functional roles of chrysin in progression of ovarian cancer cells using ES2 and OV90 (clear cell and serous carcinoma, respectively) cell lines. Results of the current study demonstrated that chrysin inhibited ovarian cancer cell proliferation and induced cell death by increasing reactive oxygen species (ROS) production and cytoplasmic Ca2+ levels as well as inducing loss of mitochondrial membrane potential (MMP). Moreover, chrysin activated mitogen-activated protein kinase (MAPK) and phosphoinositide 3-kinase (PI3K)/AKT pathways in ES2 and OV90 cells in concentration-response experiments. Collectively, our results led us to propose that chrysin-induced apoptotic events are mediated by the activation of PI3K and MAPK pathways in human ovarian cancer cells. This article is protected by copyright. All rights reserved
      PubDate: 2017-11-14T00:35:39.724286-05:
      DOI: 10.1002/jcp.26150
  • Targeting P‐glycoprotein and SORCIN: Dihydromyricetin strengthens
           anti‐proliferative efficiency of adriamycin via MAPK/ERK and
           Ca2+‐mediated apoptosis pathways in MCF‐7/ADR and K562/ADR
    • Authors: Yaoting Sun; Changyuan Wang, Qiang Meng, Zhihao Liu, Xiaokui Huo, Pengyuan Sun, Huijun Sun, Xiaodong Ma, Jinyong Peng, Kexin Liu
      Abstract: Recently, a new target Ca2+‐binding protein SORCIN was reported to participate in multidrug resistance (MDR) in cancer. Here we aim to investigate whether dihydromyricetin (DMY), a dihydroflavonol compound with anti‐inflamatory, anti‐oxidant, anti‐bacterial and anti‐tumor actions, reverses MDR in MCF‐7/ADR and K562/ADR and to elucidate its potential molecular mechanism. DMY enhanced cytotoxicity of adriamycin (ADR) by downregulating MDR1 mRNA and P‐gp expression through MAPK/ERK pathway and also inhibiting the function of P‐gp significantly. Meanwhile, DMY decreased mRNA and protein expression of SORCIN, which resulted in elevating intracellular free Ca2+. Finally, we investigated co‐administration ADR with DMY remarkably increased ADR‐induced apoptosis. Further study showed DMY elevated ROS levels and caspase‐12 protein expression, which signal apoptosis in endoplasmic reticulum. At the same time, proteins related to mitochondrial apoptosis were also changed such as Bcl‐2, Bax, caspase‐3, caspase‐9, and PARP. Finally, nude mice model also demonstrated that DMY strengthened anti‐tumor activity of ADR in vivo. In conclusion, DMY reverses MDR by downregulating P‐gp, SORCIN expression and increasing free Ca2+, as well as, inducing apoptosis in MCF‐7/ADR and K562/ADR. These fundamental findings provide evidence for further clinical research in application of DMY as an assistant agent in the treatment of cancer.When ADR co‐administrated with DMY P‐gp and SORCIN were reduced and then induced, respectively, in increasing of intracellular ADR and calcium. Finally apoptosis was triggered through mitochondria, endoplasmic reticulum pathway.
      PubDate: 2017-11-14T00:25:33.993318-05:
      DOI: 10.1002/jcp.26087
  • Protective effects of curcumin against aflatoxicosis: a comprehensive
    • Authors: Mohammad Mohajeri; Behzad Behnam, Arrigo F. G. Cicero, Amirhossein Sahebkar
      Abstract: Aflatoxicosis is a deleterious medical condition that results from aflatoxins (AFs) or ochratoxins (OTs). Contamination with these toxins exerts detrimental effects on the liver, kidneys, reproductive organs, and also on immunological and cardiovascular systems. Aflatoxicosis is closely associated with overproduction of reactive oxygen species (ROS) as key contributors to oxidative and nitrosative stress responses, and subsequent damages to lipids, proteins, RNA, and DNA. The main target organ for AF toxicity is the liver, where DNA adducts, degranulation of endoplasmic reticulum, increased hepatic lipid peroxide, GSH depletion, mitochondrial dysfunction, and reduction of enzymatic and non‐enzymatic antioxidants are manifestations of aflatoxicosis. Curcuma longa L. (turmeric) is a medicinal plant widely utilized all over the world for culinary and phytomedical purposes. Considering the antioxidant characteristic of curcumin, the main active component of turmeric, this review is intended to critically summarize the available evidence supporting possible effectiveness of curcumin against aflatoxicosis. Curcumin can serve as a promising candidate for attenuation of the adverse consequences of aflatoxicosis, acting mainly through intrinsic antioxidant effects aroused from its structure, modulation of the immune system as reflected by interleukin‐1β and transforming growth factor‐β, and interfering with AF's biotransformation by cytochrome P450 isoenzymes CYP1A, CYP3A, CYP2A, CYP2B, and CYP2C. This article is protected by copyright. All rights reserved
      PubDate: 2017-11-14T00:20:44.054362-05:
      DOI: 10.1002/jcp.26212
  • C-C Motif Chemokine Ligand 2 Regulates LPS-Induced Inflammation and ER
           Stress to Enhance Proliferation of Bovine Endometrial Epithelial Cells
    • Authors: Whasun Lim; Hyocheol Bae, Fuller W. Bazer, Sung-Man Kim, Gwonhwa Song
      Abstract: Chemokines play an important role in regulating the complex immune system at the maternal-fetal interface during pregnancy. Among various chemokines, C-C motif chemokine ligand 2 (CCL2) plays a role in the recruitment of immune regulatory cells to implantation sites within the endometrium. In cattle, CCL2 is abundantly expressed in the uterine endometrium. However, its intracellular signaling has not been identified. In this study, we examined the effects of CCL2 on bovine endometrial (BEND) cell proliferation. CCL2 stimulated BEND cell proliferation by abundant expression of PCNA, accumulation of cells in the G2/M phase, and activation of the PI3K/AKT and MAPK signaling pathways. Moreover, CCL2 reduced endoplasmic reticulum stress and restored the inflammation-induced reduction in BEND cell proliferation by regulating the unfolded protein response genes and cytokines. Collectively, these results demonstrated that CCL2 plays a pivotal role in reproductive tissues and may support maternal-fetal interface to improve efficiency of pregnancy. This article is protected by copyright. All rights reserved
      PubDate: 2017-11-10T06:30:40.837009-05:
      DOI: 10.1002/jcp.26151
  • Key‐genes regulating the liposecretion process of mature adipocytes
    • Authors: Giulia Maurizi; Tiina Petäistö, Angela Maurizi, Lucio Della Guardia
      Abstract: White mature adipocytes (MAs) are plastic cells able to reversibly transdifferentiate toward fibroblast‐like cells maintaining stem cell gene signatures. The main morphologic aspect of this transdifferentiation process, called liposecretion, is the secretion of large lipid droplets and the development of organelles necessary for exocrine secretion.There is a considerable interest in the adipocyte plastic properties involving liposecretion process, but the molecular details are incompletely explored.This review analyzes the gene expression of MAs isolated from human subcutaneous fat tissue with respect to bone marrow (BM)‐derived mesenchymal stem cells (MSC) focusing on gene regulatory pathways involved into cellular morphology changes, cellular proliferation and transports of molecules through the membrane, suggesting potential ways to guide liposecretion.In particular, Wnt, MAPK/ERK and AKT pathways were accurately described, studying up‐ and down‐stream molecules involved. Moreover, adipogenic extra‐ and intra‐cellular interactions were analyzed studying the role of CDH2, CDH11, ITGA5, E‐Syt1, PAI‐1, IGF1 and INHBB genes. Additionally, PLIN1 and PLIN2 could be key‐genes of liposecretion process regulating molecules transport through the membrane.All together data demonstrated that liposecretion is regulated through a complex molecular networks that are able to respond to microenvironment signals, cytokines and growth factors. Autocrine as well as external signaling molecules might activate liposecretion affecting adipocytes physiology. This article is protected by copyright. All rights reserved
      PubDate: 2017-11-10T06:30:22.410609-05:
      DOI: 10.1002/jcp.26188
  • Regulating Osteogenesis and Adipogenesis in Adipose‐Derived Stem Cells
           by Controlling Underlying Substrate Stiffness
    • Authors: Tao Zhang; Shiyu Lin, Xiaoru Shao, Sirong Shi, Qi Zhang, Changyue Xue, Yunfeng Lin, Bofeng Zhu, Xiaoxiao Cai
      Abstract: Cells reside in a complex microenvironment (niche) in which the biochemical and biophysical properties of the extracellular matrix profoundly affect cell behavior. Extracellular stiffness, one important bio‐mechanical characteristic of the cell niche, is important in regulating cell proliferation, migration, and lineage specification. However, the mechanism by which mechanical signals guide osteogenic and adipogenic commitment of stem cells remains difficult to dissect. To explore this question, we generated a range of polydimethylsiloxane‐based matrices with differing degrees of stiffness that mimicked the stiffness seen in natural tissues and examined adipose stem cell morphology, spreading, vinculin expression and differentiation along the osteogenic and adipogenic pathways. Rigid matrices allowed broader cell spreading, faster growth rate and stronger expression of vinculin in adipose‐derived stem cells. In the presence of inductive culture media, stiffness‐dependent osteogenesis and adipogenesis of the adipose stem cells indicated that there was a combinatorial effect of biophysical and biochemical cues; no such lineage specification was observed in normal media. Osteogenic differentiation behavior showed a correlation with matrix rigidity, as well as with elevated expression of RhoA, ROCK‐1/‐2 and related proteins in the Wnt/β‐catenin pathway. The result provides a comprehensive understanding of how stem cells respond to the surrounding microenvironment and points to the fact that matrix stiffness is a critical element in biomaterial design and this will be an important advance in stem cell‐based tissue engineering. This article is protected by copyright. All rights reserved
      PubDate: 2017-11-10T06:25:55.822566-05:
      DOI: 10.1002/jcp.26193
  • Pioglitazone inhibits cancer cell growth through STAT3 inhibition and
           enhanced AIF expression via a PPARγ‐independent pathway
    • Authors: Masanobu Tsubaki; Tomoya Takeda, Yoshika Tomonari, Keishi Kawashima, Tatsuki Itoh, Motohiro Imano, Takao Satou, Shozo Nishida
      Abstract: Pioglitazone is an anti‐diabetic agent that belongs to the thiazolidinedione class, which target peroxisome proliferator‐activated receptor γ (PPARγ), a transcription factor in the nuclear receptor family. Different cancer cells expressing high levels of PPARγ and PPARγ ligands induce cell cycle arrest, cell differentiation, and apoptosis. However, the mechanisms underlying these processes remain unknown. Here, we investigated the mechanism underlying pioglitazone‐induced apoptosis in human cancer cells. We showed that at similar concentrations, pioglitazone induced death in cancer cells expressing high or low levels of PPARγ. Combined treatment of pioglitazone and GW9662, a PPARγ antagonist, did not rescue this cell death phenotype. Z‐VAD‐fmk, a pan‐caspase inhibitor, did not reverse pioglitazone‐induced apoptosis in cancer cells expressing PPARγ at high or low levels. Pioglitazone suppressed the activation of signal transducers and activator of transcription 3 (STAT3) and Survivin expression, and enhanced the apoptosis‐inducing factor (AIF) levels in these cells. Furthermore, pioglitazone enhanced the cytotoxic effect of cisplatin and oxaliplatin by suppressing Survivin and increasing AIF expression. These results indicated that pioglitazone induced apoptosis via a PPARγ‐independent pathway, thus describing pioglitazone as a potential therapeutic agent for controlling the progression of different cancers. This article is protected by copyright. All rights reserved
      PubDate: 2017-11-10T06:25:36.778755-05:
      DOI: 10.1002/jcp.26225
  • Potential roles of metalloproteinases of endometrium‐derived exosomes in
           embryo‐maternal crosstalk during implantation
    • Authors: Zeinab Latifi; Amir Fattahi, Ali Ranjbaran, Hamid Reza Nejabati, Kazuhiko Imakawa
      Abstract: During embryo implantation, crosstalk between the endometrial epithelium and the blastocyst, especially the trophoblasts, is a prerequisite for successful implantation. During this crosstalk, various molecular and functional changes occur to promote synchrony between the embryo and the endometrium as well as the uterine cavity microenvironment. In the past few years, growing evidence has shown that endometrium‐derived exosomes play pivotal roles in the embryonic‐maternal crosstalk during implantation, although the exact mechanism of this crosstalk has yet to be determined. The presence of metalloproteinases has been reported in endometrium‐derived exosomes, implying the importance of these enzymes in exosome‐based crosstalk. Thus, in this review, we describe the potential roles of the metalloproteinases of endometrium‐derived exosomes in promoting embryo attachment and implantation. This study could provide a better understanding of the potential roles of exosomal metalloproteinases in embryo implantation and pave the way for developing novel exosome‐based regulatory agents to support early pregnancy. This article is protected by copyright. All rights reserved
      PubDate: 2017-11-08T07:00:48.221218-05:
      DOI: 10.1002/jcp.26259
  • Methylation‐independent CHFR expression is a potential biomarker
           affecting prognosis in acute myeloid leukemia
    • Authors: Jing-dong Zhou; Ting-juan Zhang, Xi-xi Li, Ji-chun Ma, Hong Guo, Xiang-mei Wen, Dong-ming Yao, Wei Zhang, Jiang Lin, Jun Qian
      Abstract: CHFR acts as a tumor suppressor gene, which is frequently inactivated caused by its promoter hypermethylation in various solid tumors. Although a recent study showed that CHFR hypermethylation was a frequent event in acute myeloid leukemia (AML) and correlated with adverse clinical outcome, herein, we found that CHFR methylation was a rare event in patients with myeloid malignancies (including AML, chronic myeloid leukemia, and myelodysplastic syndromes), but its expression may serve as an independent prognostic biomarker in AML. CHFR expression was assessed by real‐time quantitative PCR, whereas CHFR methylation was detected by methylation‐specific PCR and bisulfite sequencing PCR. In AML patients, lower CHFR expression was associated with lower complete remission (CR) rate, and CHFR expression was significantly increased in CR after chemotherapy. Moreover, patients with lower CHFR expression showed shorter overall survival and leukemia‐free survival, and multivariate analysis confirmed that lower CHFR expression was an independent risk factor in AML. Importantly, the prognostic value of CHFR expression was validated using the published Gene Expression Omnibus datasets. Notably, CHFR promoter was nearly unmethylated in patients with myeloid malignancies. Our findings revealed that lower CHFR expression was independently associated with unfavorable prognosis in AML. Moreover, aberrant CHFR promoter methylation was a rare event in myeloid malignances. This article is protected by copyright. All rights reserved
      PubDate: 2017-11-08T06:56:25.331102-05:
      DOI: 10.1002/jcp.26253
  • Effects of XIST/miR‐137 axis on neuropathic pain by targeting
           TNFAIP1 in a rat model
    • Authors: Ying Zhao; Sen Li, Min Xiao, Yan Shi, Chang-Ming Zhao
      Abstract: Non‐coding RNAs have been reported to participate in the pathophysiology of neuropathic pain. The objective of our study was to investigate the biological role of XIST in neuropathic pain development. In our study, we identify and validate that lncRNA XIST was markedly increased and miR‐137 was significantly decreased in chronic constriction injury (CCI) rats. XIST silencing alleviated pain behaviors including both mechanical and thermal hyperalgesia in the CCI rats. XIST was predicted to interact with miR‐137 by bioinformatics technology and dual‐luciferase reporter assays confirmed the correlation between XIST and miR‐137. miR‐137 was negatively modulated by XIST and upregulation of miR‐137 greatly reduced neuropathic pain development in CCI rats. Moreover, we observed that tumor necrosis factor alpha‐induced protein 1 (TNFAIP1) was enhanced in CCI rats and 3'‐untranslated region (UTR) of TNFAIP1 was exhibited to be a target of miR‐137 by bioinformatics prediction. TNFAIP1 can act as a crucial inflammation regulator by activating NF‐kB activity. Overexpression of miR‐137 significantly suppressed TNFAIP1 both in vitro and in vivo. Furthermore, upregulation of XIST reversed the inhibitory role of miR‐137 in neuropathic pain development by inhibiting TNFAIP1. In conclusion, our current study indicates that XIST can positively regulate neuropathic pain in rats through regulating the expression of miR‐137 and TNFAIP1. Our results imply that XIST/miR‐137/TNFAIP1 axis may serve as a novel therapeutic target in neuropathic pain. This article is protected by copyright. All rights reserved
      PubDate: 2017-11-08T06:56:01.062641-05:
      DOI: 10.1002/jcp.26254
  • Fatty Acid Elongase‐7 Is Regulated Via SP1 and Is Involved in Lipid
           Accumulation in Bovine Mammary Epithelial Cells
    • Authors: Si Chen; Zhigang Hu, Hua He, Xiaolin Liu
      Abstract: Fatty Acid Elongase 7 (ELOVL7) is the newly discovered protein on human that catalyzes the rate‐limiting step towards the synthesis of very long‐chain fatty acids and exhibits the highest activity toward C18:3 (n‐3) acyl‐CoAs, which is the precursor of eicosapentaenoic acid (EPA, 20:5n‐3). However, in ruminants, an overall understanding of ELOVLs gene family and the transcriptional regulation of ELOVL7 remain unknown. The purpose of this study is to investigate the transcriptional regulation and the influence of bovine ELOVL7 in bovine mammary epithelial cells (bMECs). Quantitative real‐time PCR analysis demonstrated that ELOVLs gene family had differential expression patterns in bMECs, and bovine ELOVL7 was expressed in a tissue‐specific manner, which was high in kidney, followed by in abdominal fat and in bMECs. Promoter analysis of bovine ELOVL7, including bioinformatics analyses, dual‐luciferase reporter assays, protein pull‐down assay, western bolt assay, over expression and RNA interference assay, have independently and synthetically demonstrated that transcription factor Sp1 (SP1) specifically interacted with the GC‐box at ‐143 to ‐128 base pair on ELOVL7 promoter. Furthermore, the exogenous α‐linolenic acid (ALA, 18:3n‐3), strengthened the binding of SP1 to the ELOVL7 proximal promoter, resulting in the accumulation of lipid droplets in bMECs. In conclusion, these data suggest that the transcription of bovine ELOVL7 are affected by the binding of SP1 and the treatment of ALA , moreover, enlightening us the profound role of SP1 in modulating lipid synthesis of the mammary gland in cattle. This article is protected by copyright. All rights reserved
      PubDate: 2017-11-08T06:55:57.412155-05:
      DOI: 10.1002/jcp.26255
  • Endogenous annexin A1 (AnxA1) modulates early phase gestation and
           offspring sex‐ratio skewing
    • Authors: Cristina Bichels Hebeda; Isabel Daufenback Machado, Isadora Reif-Silva, Jusciele Brogin Moreli, Sonia Maria Oliani, Suchita Nadkarni, Mauro Perretti, Estela Bevilacqua, Sandra H. P. Farsky
      Abstract: Annexin A1 (AnxA1) is a glucocorticoid‐regulated anti‐inflammatory protein secreted by phagocytes and other specialised cells. In the endocrine system, AnxA1 controls secretion of steroid hormones and it is abundantly expressed in the testis, ovaries, placenta and seminal fluid, yet its potential modulation of fertility has not been described. Here we observed that AnxA1 knockout (KO) mice delivered a higher number of pups, with a higher percentage of female offsprings. This profile was not dependent on the male features, as sperm from KO male mice did not present functional alterations, and had an equal proportion of Y and X chromosomes, comparable to wild type (WT) male mice. Furthermore, mismatched matings of male WT mice with female KO yielded a higher percentage of female pups per litter, a phenomenon which was not observed when male KO mice mated with female WT animals. Indeed, AnxA1 KO female mice displayed several differences in parameters related to gestation including i) an arrested estrous cycle at proestrus phase, ii) increased sites of implantation, iii) reduced pre‐ and post‐implantation losses, iv) exacerbated features of the inflammatory reaction in the uterine fluid during implantation phase and v) enhanced plasma progesterone in the beginning of pregnancy. In summary, herein we highlight that AnxA1 pathway as a novel determinant of fundamental non‐redundant regulatory functions during early pregnancy. This article is protected by copyright. All rights reserved
      PubDate: 2017-11-08T06:55:49.630612-05:
      DOI: 10.1002/jcp.26258
  • MiR‐26a promoted endometrial epithelium cells (EECs) proliferation and
           induced stromal cells (ESCs) apoptosis via the PTEN‐PI3K/AKT pathway in
           dairy goats
    • Authors: Lei Zhang; Xiaorui Liu, Junze Liu, Xingna Ma, Zhanqin Zhou, Yuxuan Song, Binyun Cao
      Abstract: Changes in endometrial cell morphology and function are absolutely necessary for successful embryo implantation. In this study, miR‐26a was widely expressed in dairy goats, and was found to be regulated by β‐estradiol (E2) and progesterone (P4) in endometrial epithelium cells (EECs) as well as stromal cells (ESCs). Furthermore, miR‐26a played a role in the regulation of cells proliferation and apoptosis by directly regulating PTEN and indirectly regulating the PI3K/AKT pathway in EECs but not in ESCs of dairy goats in vitro. In addition, miR‐26a regulated the expression of osteopontin (OPN), vascular endothelial growth factor (VEGF), Cyclooxygenase‐2 (COX‐2) and prolactin (PRL) in endometrial cells. Therefore, we could get a conclusion that miR‐26a had very complex and diverse functions in the endometrial cells during the development of endometrial receptivity in dairy goats. This study provided an efficient platform for studying the regulatory effect of miR‐26a on endometrial cells during the development of endometrial receptivity in dairy goats. This article is protected by copyright. All rights reserved
      PubDate: 2017-11-08T06:55:47.618983-05:
      DOI: 10.1002/jcp.26252
  • Four differentially methylated gene pairs to predict the prognosis for
           early stage hepatocellular carcinoma patients
    • Authors: Shaoguang Liu; Changfeng Miao, Juan Liu, Chang-Cheng Wang, Xiao-Jie Lu
      Abstract: Hepatocellular carcinoma (HCC) was the second most common malignant tumor with a poor prognostic condition. We aimed to identify novel methylation signatures to predict HCC patients at their early stages. Differentially expressed methylated genes between HCC patients and normal liver tissues retrieved from TCGA were screened out by SAM. Genes highly related to patients' survival were figured out by COX. The signatures that could identify relapse HCC patients were identified by the forwarding search algorithm. Besides, functional enrichment analysis was performed on the methylation genes in the signature. A total of 5392 CPG sites that differentially methylated expressed were found out and 4294 differentially expressed genes were obtained. A total of 197 genes among were associated with RFS of HCC patients at both stage I and stage II. Signature composed of 21 pairs was obtained to predict the prognostic situation by C‐index forward search method. The function of these genes was figured out by functional enrichment analysis. To summary, the signature composed of 21 gene pairs that can predict the prognostic situation of HCC patients at both stage I and stage II, provided a reference standard for the adjuvant therapy of HCC patients after surgery. This article is protected by copyright. All rights reserved
      PubDate: 2017-11-08T06:55:23.335542-05:
      DOI: 10.1002/jcp.26256
  • Glucose stimulates intestinal epithelial crypt proliferation by modulating
           cellular energy metabolism
    • Authors: Weinan Zhou; Deepti Ramachandran, Abdelhak Mansouri, Megan J. Dailey
      Abstract: The intestinal epithelium plays an essential role in nutrient absorption, hormone release and barrier function. Maintenance of the epithelium is driven by continuous cell renewal by stem cells located in the intestinal crypts. The amount and type of diet influence this process and result in changes in the size and cellular make‐up of the tissue. The mechanism underlying the nutrient‐driven changes in proliferation is not known, but may involve a shift in intracellular metabolism that allows for more nutrients to be used to manufacture new cells. We hypothesized that nutrient availability drives changes in cellular energy metabolism of small intestinal epithelial crypts that could contribute to increases in crypt proliferation. We utilized primary small intestinal epithelial crypts from C57BL/6J mice to study 1) the effect of glucose on crypt proliferation, and 2) the effect of glucose on crypt metabolism using an extracellular flux analyzer for real‐time metabolic measurements. We found that glucose increased both crypt proliferation and glycolysis, and the glycolytic pathway inhibitor 2‐Deoxy‐D‐glucose (2‐DG) attenuated glucose‐induced crypt proliferation. Glucose did not enhance glucose oxidation, but did increase the maximum mitochondrial respiratory capacity, which may contribute to glucose‐induced increases in proliferation. Glucose activated Akt/HIF‐1α signaling pathway, which might be at least in part responsible for glucose‐induced glycolysis and cell proliferation. These results suggest that high glucose availability induces an increase in crypt proliferation by inducing an increase in glycolysis with no change in glucose oxidation. This article is protected by copyright. All rights reserved
      PubDate: 2017-11-01T08:10:51.248349-05:
      DOI: 10.1002/jcp.26199
  • Downregulation of P2Y12 in the superior cervical ganglia alleviates
           abnormal sympathetic activity after myocardial ischemia
    • Authors: Lifang Zou; Yingxin Gong, Shanhong Zhao, Zhihua Yi, Xinyao Han, Bing Wu, Tianyu Jia, Lin Li, Huilong Yuan, Liran Shi, Chunping Zhang, Yun Gao, Guilin Li, Hong Xu, Hui Liu, Shangdong Liang, Shuangmei Liu
      Abstract: Background/Aims: Superior cervical ganglia (SCG) innervate the myocardium and participate in sympathoexcitatory transmission. P2Y12 receptor is expressed in satellite glial cells (SGCs). This study seeks to clarify whether the P2Y12 receptor is involved in the sympathoexcitation reflex after myocardial ischemia (MI). Methods: MI model was induced by occlusion of the left coronary artery. P2Y12 were assayed by real time PCR and Western blotting. Results: Our results showed that expression levels of P2Y12 mRNA and protein were significantly higher in the MI group than in the sham group. Administration of P2Y12 short hairpin RNA (shRNA) caused downregulation of the P2Y12 receptor in the SCG. In MI rats plus P2Y12 shRNA treatment group, the abnormal changes in diastolic blood pressure (DBP), systolic blood pressure (SBP), heart rate (HR), electrocardiograms (ECGs), and cardiac tissue structures were alleviated. When the treatment of P2Y12 shRNA in MI rats, upregulated co‐expression values of P2Y12 and glial fibrillary acidic protein (GFAP), the upregulation of tumor necrosis factor α (TNF‐α) and phosphorylated P38 mitogen activated protein kinase (p‐P38 MAPK) in the SCG were decreased. Conclusion: Downregulation of the P2Y12 receptor in the SCG after MI may improve cardiac function by alleviating the sympathoexcitatory reflex. This article is protected by copyright. All rights reserved
      PubDate: 2017-11-01T08:10:42.588238-05:
      DOI: 10.1002/jcp.26184
    • Authors: Flávia de T. Frias; Karina C.e. Rocha, Mariana de Mendonça, Gilson M. Murata, Hygor N. Araujo, Luís G.O. de Sousa, Érica de Sousa, Sandro M. Hirabara, Nayara de C. Leite, Everardo M. Carneiro, Rui Curi, Leonardo R. Silveira, Alice C. Rodrigues
      Abstract: The effect of fenofibrate on the metabolism of skeletal muscle and visceral white adipose tissue of diet‐induced obese (DIO) mice was investigated. C57BL/6J male mice were fed either a control or high‐fat diet for eight weeks. Fenofibrate (50 mg/Kg b.w., daily) was administered by oral gavage during the last two weeks of the experimental period. Insulin‐stimulated glucose metabolism in soleus muscles, glucose tolerance test, insulin tolerance test, indirect calorimetry, lipolysis of visceral white adipose tissue, expression of miR‐103‐3p in adipose tissue and miR‐1a, miR‐133a/b, miR‐206, let7b‐5p, miR‐23b‐3p, miR‐29‐3p, miR‐143‐3p in soleus muscle, genes related to glucose and fatty acid metabolism in adipose tissue and soleus muscle, and proteins (phospho‐AMPKα2, Pgc1α, Cpt1b), intramuscular lipid staining, and activities of fatty acid oxidation enzymes in skeletal muscle were investigated. In DIO mice, fenofibrate prevented weight gain induced by HFD feeding by increasing energy expenditure; improved whole body glucose homeostasis, and in skeletal muscle, increased insulin dependent glucose uptake, miR‐1a levels, reduced intramuscular lipid accumulation, and phospho‐AMPKα2 levels. In visceral adipose tissue of obese mice, fenofibrate decreased basal lipolysis rate and visceral adipocytes hypertrophy, and induced the expression of Glut‐4, Irs1 and Cav‐1 mRNA and miR‐103‐3p suggesting a higher insulin sensitivity of the adipocytes. The evidence is presented herein that beneficial effects of fenofibrate on body weight, glucose homeostasis and muscle metabolism might be related to its action in adipose tissue. Moreover, fenofibrate regulates miR‐1a‐3p in soleus and miR‐103‐3p in adipose tissue, suggesting these microRNAs might contribute to fenofibrate beneficial effects on metabolism. This article is protected by copyright. All rights reserved
      PubDate: 2017-11-01T08:06:13.047536-05:
      DOI: 10.1002/jcp.26203
  • Efficacy of different chemotherapy regimens in treatment of advanced or
           metastatic pancreatic cancer: a network meta‐analysis
    • Authors: Shu-Hua Zhang; Gui-Feng Liu, Xue-Feng Li, Lin Liu, Shao-Nan Yu
      Abstract: We performed a network meta‐analysis (NMA) to compare the short‐ and long‐term efficacy of Gemcitabine, Gemcitabine + S‐1 (tegafur), Gemcitabine + nab‐paclitaxel, Gemcitabine + Capecitabine, Gemcitabine + Cisplatin, FOLFIRINOX (oxaliplatin + irinotecan + fluorouracil + leucovorin), Gemcitabine + oxaliplatin, Gemcitabine + irinotecan, Gemcitabine + Exatecan, Gemcitabine + pemetrexed, Gemcitabine + 5‐FU and S‐1 in treating advanced or metastatic pancreatic cancer (PC). The odds radios (OR) or weighted mean difference (WMD) and surface under the cumulative ranking curves (SUCRA) were evaluated by a combination of direct evidence and indirect evidence. In total twenty studies were included in this paper. For short‐term efficacy, the overall response rate (ORR) was lower for patients treated with Gemcitabine compared with Gemcitabine + S‐1, Gemcitabine + Cisplatin, Gemcitabine + irinotecan and S‐1. The ORR for FOLFIRINOX was higher compared with Gemcitabine, Gemcitabine + Capecitabine and Gemcitabine + Cisplatin. The disease control rate (DCR) for Gemcitabine was lower compared with Gemcitabine + S‐1, Gemcitabine + Cisplatin and FOLFIRINOX. For long‐term efficacy, the 12‐month overall survival (OS) rate for FOLFIRINOX was higher compared with Gemcitabine, Gemcitabine + Capecitabine, Gemcitabine + Cisplatin, Gemcitabine + irinotecan, Gemcitabine + Exatecan and Gemcitabine + pemetrexed. The SUCRA revealed that FOLFIRINOX was relatively better in both short‐term and long‐term efficacy, while Gemcitabine was relatively poorer. In both short‐term and long‐term efficacy, FOLFIRINOX had the best short term and long term efficacy among the 12 chemotherapy regimens while efficacy of Gemcitabine was relatively poorer in the treatment of advanced or metastatic PC. This article is protected by copyright. All rights reserved
      PubDate: 2017-11-01T08:05:38.499585-05:
      DOI: 10.1002/jcp.26183
    • Authors: Jon Ramsey; Kelly Butnor, Zhihua Peng, Tim Leclair, Jos van der Velden, Gary Stein, Jane Lian, C. Matthew Kinsey
      Abstract: The mammalian runt‐related factor 1 (RUNX1) is a master transcription factor that regulates lineage specification of hematopoietic stem cells. RUNX1 translocations result in the development of myeloid leukemias. Recently, RUNX1 has been implicated as a tumor suppressor in other cancers. We postulated RUNX1 expression may be associated with lung adenocarcinoma etiology and/or progression. We evaluated the association of RUNX1 mRNA expression with overall survival data from The Cancer Genome Atlas (TCGA), a publically available database. Compared to high expression levels, Low RUNX1 levels from lung adenocarcinomas were associated with a worse overall survival (Hazard Ratio = 2.014 (1.042 to 3.730 95% confidence interval), log‐rank P = 0.035) compared to those that expressed high RUNX1 levels. Further immunohistochemical examination of 85 surgical specimens resected at the University of Vermont Medical Center identified that low RUNX1 protein expression was associated with larger tumors (P = 0.038). Gene expression network analysis was performed on the same subset of TCGA cases that demonstrated differential survival by RUNX1 expression. This analysis, which reveals regulatory relationships, showed that reduced RUNX1 levels were closely linked to upregulation of the transcription factor E2F1. To interrogate this relationship, RUNX1 was depleted in a lung cancer cell line that expresses high levels of RUNX1. Loss of RUNX1 resulted in enhanced proliferation, migration, and invasion. RUNX1 depletion also resulted in increased mRNA expression of E2F1 and multiple E2F1 target genes. Our data implicate loss of RUNX1 as driver of lung adenocarcinoma aggression, potentially through deregulation of the E2F1 pathway. This article is protected by copyright. All rights reserved
      PubDate: 2017-11-01T08:00:46.494403-05:
      DOI: 10.1002/jcp.26201
  • Substrate stiffness regulated migration and angiogenesis potential of A549
           cells and HUVECs
    • Authors: Dan Zhao; Changyue Xue, Qianshun Li, Mengting Liu, Wenjuan Ma, Tengfei Zhou, Yunfeng Lin
      Abstract: Tumor tissue tends to stiffen during solid tumor progression. Substrate stiffness is known to alter cell behaviors, such as proliferation and migration, during which angiogenesis is requisite. Mono‐culture and co‐culture systems of lung cancer cell line A549 and human umbilical vein endothelial cells (HUVECs), on polydimethylsiloxane substrates (PDMS) with varying stiffness, were used for investigating the effects of substrate stiffness on the migration and angiogenesis of lung cancer. The expressions of matrix metalloproteinases (MMPs) and angiogenesis‐related growth factors were up‐regulated with the increase of substrate stiffness, whereas that of tissue inhibitor of matrix metalloproteinase (TIMPs) were down‐regulated with increasing substrate stiffness. Our data not only suggested that stiff substrate may promote the migration and angiogenesis capacities of lung cancer, but also suggested that therapeutically targeting lung tumor stiffness or response of ECs to lung tumor stiffness may help reduce migration and angiogenesis of lung tumor. This article is protected by copyright. All rights reserved
      PubDate: 2017-10-27T05:02:15.272281-05:
      DOI: 10.1002/jcp.26189
  • Pulmonary Hypertension: Molecular Aspects of Current Therapeutic
           Intervention and Future Direction
    • Authors: Taruna K. Arora; Amit K. Arora, Monika K. Sachdeva, Satyendra K. Rajput, Arun K. Sharma
      Abstract: Pulmonary hypertension (PH) is a life‐threatening lung disorder with towering prevalence and risk for future has been gradually rising worldwide. Even, no specific medications are available for pulmonary hypertension; various classes of treatment based upon the origin and magnitude of hypertension are still used for the treatment of PH. Consideration of molecular or signaling modulation is the imperative approach that can offer a new notion for prevalent pharmacotherapeutic agents. Instead of concurrent targets, including endothelin receptor antagonists (ETA/ETB), phosphodiesterase 5 inhibitor (PDF‐5), calcium channel blockers, anticoagulants, diuretics and long acting prostacyclin analogue, recent scientific reports revealed the numerous potential alternative therapeutic approaches that can significantly target the pathological signaling alteration associated with PH. Understanding precise molecular cascade involved in PH can be useful for designing preclinical animal experiments and human clinical trials to evaluate target specific novel therapeutic interventions for the treatment of PH. In this review, we discussed the possible molecular signaling involved in the pathogenesis of PH and detailed account of the current status of medications employed for the treatment of PH. Moreover, the newly identified potential target sites and alternative approaches for treating the PH have been discussed. This article is protected by copyright. All rights reserved
      PubDate: 2017-10-27T04:56:01.423562-05:
      DOI: 10.1002/jcp.26191
  • FAM20C regulates osteoblast behaviors and intracellular signaling pathways
           in a cell‐autonomous manner
    • Authors: Chao Liu; Hua Zhang, Priyam Jani, Xiaofang Wang, Yongbo Lu, Nan Li, Jing Xiao, Chunlin Qin
      Abstract: Recent studies indicate that Family with sequence similarity 20 member C (FAM20C) catalyzes the phosphorylation of secreted proteins, and participates in a variety of biological processes, including cell proliferation, migration, mineralization and phosphate homeostasis. To explore the local influences of FAM20C on osteoblast, Fam20c‐deficient osteoblasts were generated by treating the immortalized Fam20cf/f osteoblasts with CMV‐Cre‐IRES‐EGFP lentivirus. Compared with the normal Fam20cf/f osteoblasts, the expression of Bone sialoprotein (Bsp), Osteocalcin (Ocn), Fibroblast growth factor 23 (Fgf23) and transcription factors that promote osteoblast maturation were up‐regulated in the Fam20c‐deficient osteoblasts. In contrast, the expression of Dental matrix protein 1 (Dmp1), Dentin sialophosphoprotein (Dspp), Osteopontin (Opn), type I Collagen a 1 (Col1a1) and Alkine phosphatase (Alp) were down‐regulated in the Fam20c‐deficient cells. These alterations disclosed the primary regulation of Fam20c on gene expression. The Fam20c‐deficient osteoblasts showed a remarkable reduction in the ability of forming mineralized nodules. However, supplements of extracellular matrix proteins extracted from the normal bone failed to rescue the reduced mineralization, suggesting that FAM20C may affect the biomineralization by the means more than local phosphorylation of extracellular matrix proteins and systemic phosphorus homeostasis. Moreover, although Fam20c deficiency had little impact on cell proliferation, it significantly reduced cell migration and lowered the levels of p‐Smad1/5/8, p‐Erk and p‐p38, suggesting that the kinase activity of FAM20C might be essential to cell mobility and the activity of BMP ligands. In summary, these findings provide evidences that FAM20C may regulate osteoblast maturation, migration, mineralization and BMP signaling pathways in a cell‐autonomous manner. This article is protected by copyright. All rights reserved
      PubDate: 2017-10-27T04:53:58.894436-05:
      DOI: 10.1002/jcp.26200
  • Gap Junctional Intercellular Communication in Adipose‐derived
           Stromal/Stem Cells is Cell Density‐dependent and Positively Impacts
           Adipogenic Differentiation
    • Authors: Miriam Wiesner; Oliver Berberich, Christiane Hoefner, Torsten Blunk, Petra Bauer-Kreisel
      Abstract: Adipose‐derived stromal/stem cells (ASCs) represent a widely used cell source with multi‐lineage differentiation capacity in approaches for tissue engineering and regenerative medicine. Despite the multitude of literature on their differentiation capacity, little is reported about the physiological properties contributing to and controlling the process of lineage differentiation. Direct intercellular communication between adjacent cells via gap junctions has been shown to modulate differentiation processes in other cell types, with connexin 43 (Cx43) being the most abundant isoform of the gap junction‐forming connexins. Thus, in the present study we focused on the expression of Cx43 and gap junctional intercellular communication (GJIC) in human ASCs, and its significance for adipogenic differentiation of these cells.Cx43 expression in ASCs was demonstrated histologically and on the gene and protein expression level, and was shown to be greatly positively influenced by cell seeding density. Functionality of gap junctions was proven by dye transfer analysis in growth medium. Adipogenic differentiation of ASCs was shown to be also distinctly elevated at higher cell seeding densities. Inhibition of GJIC by 18α‐glycyrrhetinic acid (AGA) significantly compromised adipogenic differentiation, as demonstrated by histology, triglyceride quantification, and adipogenic marker gene expression. Flow cytometry analysis showed a lower proportion of cells undergoing adipogenesis when GJIC was inhibited, further indicating the importance of GJIC in the differentiation process.Altogether, this study demonstrates the impact of direct cell‐cell communication via gap junctions on the adipogenic differentiation process of ASCs, and may contribute to further integrate direct intercellular crosstalk in rationales for tissue engineering approaches. This article is protected by copyright. All rights reserved
      PubDate: 2017-10-27T04:50:58.009568-05:
      DOI: 10.1002/jcp.26178
  • Oroxyloside inhibits angiogenesis through suppressing internalization of
           VEGFR2/Flk‐1 in endothelial cells
    • Authors: Kai Zhao; Xiaorui Li, Binyan Lin, Dawei Yang, Yuxin Zhou, Zhiyu Li, Qinglong Guo, Na Lu
      Abstract: Increasing flavonoids have been reported to possess anti‐angiogenic effects. Inhibition of angiogenesis plays a critical role in the treatment of cancer, especially in advanced metastatic cancer. In this study, we assessed the effect of Oroxylin A‐7‐glucuronide (Oroxyloside), a main metabolite of Oroxylin A, on angiogenesis in human endothelial cell‐like EA.hy926 cells. Oroxyloside suppressed the migration and tube formation of EA.hy926 cells. Meanwhile, microvessels sprouting from aortic rings and new blood vessels on the chicken chorioallantoic membrane (CAM) were also inhibited. Mechanism studies showed that Oroxyloside reduced the autophosphorylation of vascular endothelial growth factor receptor 2 (VEGFR2/Flk‐1) while it up‐regulated the expression of R‐Ras and VE‐cadherin. In consequence, Oroxyloside inhibited the downstream Akt/MAPK/NF‐κB pathways and then decreased the nuclear translocation and DNA binding ability of NF‐κB. Furthermore, in‐vivo study showed that Oroxyloside exhibited a potential anti‐angiogenic effect in Matrigel plug assay and inhibited growth of xenografted tumors with low systemic toxicity, which could be ascribed to the inhibition of VEGFR2 internalization. Taken together, these results suggested that Oroxyloside could inhibit angiogenesis in vitro and in vivo via suppressing the internalization of VEGFR2 and might serve as a potential antitumor agent. This article is protected by copyright. All rights reserved
      PubDate: 2017-10-27T04:42:39.41833-05:0
      DOI: 10.1002/jcp.26198
  • Suppressing Angiogenesis Regulates the Irradiation‐induced Stimulation
           on Osteoclastogenesis in vitro
    • Authors: Ling Tong; Guoying Zhu, Jianping Wang, Ruilian Sun, Feilong He, Jianglong Zhai
      Abstract: Ionizing radiation‐induced bone loss is a potential health concern in radiotherapy, occupational exposure and astronauts. Although impaired bone vasculature and reduced proliferation of bone‐forming osteoblasts has been implicated in this process, it has not been clearly characterized that whether radiation affects the growth of bone‐resorbing osteoclasts. The molecular crosstalk between different cell populations in the skeletal system has not yet been elucidated in detail, especially between the increased bone resorption at early stage of post‐irradiation and bone marrow‐derived endothelial progenitor cells (BM‐EPCs). In order to further understand the mechanisms involved in radiation‐induced bone loss at the cellular level, we assessed the effects of irradiation on angiogenesis of BM‐EPCs and osteoclastogenesis of receptor activator for nuclear factor‐κB ligand (RANKL)‐stimulated RAW 264.7 cells and crosstalk between these cell populations. We herein found significantly dysfunction of BM‐EPCs in response to irradiation at a dose of 2 Gy, including inhibited proliferation, migration, tube‐forming abilities and downregulated expression of pro‐angiogenesis vascular endothelial growth factors A (VEGF A). Meanwhile, we observed that irradiation promoted osteoclastogenesis of RANKL‐stimulated RAW 264.7 cells directly or indirectly. These results provide quantitative evidences of irradiation induced osteoclastogenesis at a cellular level, and strongly suggest the involvement of osteoclastogenesis, angiogenesis and crosstalk between bone marrow cells in the radiation‐induced bone loss. This study may provide new insights for the early diagnosis and intervention of bone loss post‐irradiation.This article is protected by copyright. All rights reserved
      PubDate: 2017-10-27T04:39:14.302403-05:
      DOI: 10.1002/jcp.26196
  • Role of Viruses in Gastrointestinal cancer
    • Authors: Habibollah Mirzaei; Hossein Goudarzi, Gita Eslami, Ebrahim Faghihloo
      Abstract: Gastrointestinal cancers are a global public health problem, which represent a vast majority of all cancer‐caused deaths in both men and women. On the other hand, viral pathogens have been long implicated as etiological factors in the onset of certain human cancers, including gastrointestinal tumors. In this regard, Human Papilloma Virus (HPV), Epstein ‐ Barr Virus (EBV) and John Cunningham Virus (JCV) have been more strongly suggested to be involved in gastrointestinal carcinogenesis; so that, the association of HPV with oropharyngeal and anal cancers and also the association of EBV with gastric cancer have been etiologically confirmed by epidemiological and experimental investigations. Although, the association of other viruses is less evident, but may rely on co‐factors for their oncogenic roles. Therefore, to improve the prevention and treatment of these classes of cancer, their association with viral agents as potential risk factors should be investigated with care. In this respect, the present review has focused on the existing literature on the subject of viral involvement in gastrointestinal tumorgenesis, by covering and discussing various gastrointestinal cancers, corresponding viral agents and their oncogenic aspects and then summarizing evidences either supporting or rejecting a causal role of these pathogens in gastrointestinal malignancies. This article is protected by copyright. All rights reserved
      PubDate: 2017-10-27T04:38:09.098691-05:
      DOI: 10.1002/jcp.26194
  • Long non‐coding RNA LINC00968 acts as oncogene in NSCLC by activating
           the Wnt signaling pathway
    • Authors: Yíng Wang; Jun Zhou, Yu-Jun Xu, Hai-Bo Hu
      Abstract: Long non‐coding RNAs (lncRNAs) have played critical roles in a variety of cancers, including non‐small cell lung cancer (NSCLC). In our study, we focused on the biological function and clinical significance of lncRNA LINC00968 in NSCLC. It was indicated that LINC00968 was significantly increased in LUAD tissues, LUSC tissues and NSCLC cells compared to their corresponding controls. Inhibition of LINC00968 was able to repress NSCLC growth, migration and invasion in vitro while upregulation of LINC00968 reversed this process. Additionally, downregulation of LINC00968 induced apoptosis capacity of A549 cell. Apoptosis‐related proteins BCL‐2 were decreased and BAX was increased by knockdown of LINC00968 respectively. Meanwhile we observed that Wnt signaling pathway was involved in the LINC00968‐induced NSCLC progression. Finally, in vivo tumor xenografts were established using A549 cells to detect the function of LINC00968 in NSCLC tumorigenesis. Silencing LINC00968 greatly inhibited NSCLC tumor progression, which was consistent with the in vitro tests. In conclusion, we have uncovered that LINC00968 could be regarded as a novel prognostic biomarker and therapeutic target in NSCLC diagnosis and treatment. This article is protected by copyright. All rights reserved
      PubDate: 2017-10-27T04:32:49.907929-05:
      DOI: 10.1002/jcp.26186
  • PACAP and PAC1R are differentially expressed in motor cortex of
           amyotrophic lateral sclerosis patients and support survival of
           iPSC‐derived motor neurons
    • Authors: Gabriele Bonaventura; Rosario Iemmolo, Agata G. D'Amico, Valentina La Cognata, Erminio Costanzo, Mario Zappia, Velia D'Agata, Francesca L. Conforti, Eleonora Aronica, Sebastiano Cavallaro
      Abstract: Amyotrophic lateral sclerosis (ALS) is a fatal and disabling neurodegenerative disease characterized by upper and lower motor neurons depletion. In our previous work, comprehensive genomic profiling of 41 motor cortex samples enabled to discriminate controls from sporadic ALS patients, and segregated these latter into two distinct subgroups (SALS1 and SALS2), each associated with different deregulated genes. In the present study, we focused our attention on two of them, Pituitary Adenylate Cyclase‐Activating Polypeptide (PACAP) and its type 1 receptor (PAC1R), and validated the results of the transcriptome experiments by quantitative reverse transcription‐polymerase chain reaction (qRT‐PCR), immunohistochemistry and western blot analysis. To assess the functional role of PACAP and PAC1R in ALS, we developed an in vitro model of human induced Pluripotent Stem Cells (iPSC)‐derived motor neurons and examined the trophic effects of exogenous PACAP following neurodegenerative stimuli. Treatment with 100 nM PACAP was able to effectively rescue iPSC‐derived motor neurons from apoptosis, as shown by cell viability assay and protein dosage of the apoptotic marker (BAX). All together, these data suggest that perturbations in the PACAP‐PAC1R pathway may be involved in ALS pathology and represent a potential drug target to enhance motor neuron viability. This article is protected by copyright. All rights reserved
      PubDate: 2017-10-20T03:50:35.024095-05:
      DOI: 10.1002/jcp.26182
  • Functional Role of Mesenchymal Stem Cells in the Treatment of Chronic
           Neurodegenerative Diseases
    • Authors: Debora Lo Furno; Giuliana Mannino, Rosario Giuffrida
      Abstract: Mesenchymal stem cells (MSCs) can differentiate into not only cells of mesodermal lineages, but also into endodermal and ectodermal derived elements, including neurons and glial cells. For this reason, MSCs have been extensively investigated to develop cell‐based therapeutic strategies, especially in pathologies whose pharmacological treatments give poor results, if any. As in the case of irreversible neurological disorders characterized by progressive neuronal death, in which behavioral and cognitive functions of patients inexorably decline as the disease progresses. In this review, we focus on the possible functional role exerted by MSCs in the treatment of some disabling neurodegenerative disorders such as Alzheimer's Disease, Amyotrophic Lateral Sclerosis, Huntington's Disease and Parkinson's Disease. Investigations have been mainly performed in vitro and in animal models by using MSCs generally originated from umbilical cord, bone marrow or adipose tissue. Positive results obtained have prompted several clinical trials, the number of which is progressively increasing worldwide. To date, many of them have been primarily addressed to verify the safety of the procedures but some improvements have already been reported, fortunately. Although the exact mechanisms of MSC‐induced beneficial activities are not entirely defined, they include neurogenesis and angiogenesis stimulation, antiapoptotic, immunomodulatory and anti‐inflammatory actions. Most effects would be exerted through their paracrine expression of neurotrophic factors and cytokines, mainly delivered at damaged regions, given the innate propensity of MSCs to home to injured sites. Hopefully, in the near future more efficacious cell‐replacement therapies will be developed to substantially restore disease‐disrupted brain circuitry. This article is protected by copyright. All rights reserved
      PubDate: 2017-10-20T03:35:32.440098-05:
      DOI: 10.1002/jcp.26192
  • Butyrate stimulates the growth of human intestinal smooth muscle cells by
           activation of Yes-Associated Protein
    • Authors: Li-Na Dai; Jun-Kai Yan, Yong-Tao Xiao, Jie Wen, Tian Zhang, Ke-Jun Zhou, Yang Wang, Wei Cai
      Abstract: Intestinal smooth muscle cells play a critical role in the remodeling of intestinal structure and functional adaptation after bowel resection. Recent studies have shown that supplementation of butyrate (Bu) contributes to the compensatory expansion of a muscular layer of the residual intestine in a rodent model of short-bowel syndrome (SBS). However, the underlying mechanism remains elusive. In this study, we found that the growth of human intestinal smooth muscle cells (HISMCs) was significantly stimulated by Bu via activation of Yes-Associated Protein (YAP). Incubation with 0.5 mM Bu induced a distinct proliferative effect on HISMCs, as indicated by the promotion of cell cycle progression and increased DNA replication. Notably, YAP silencing by RNA interference or its specific inhibitor significantly abolished the proliferative effect of Bu on HISMCs. Furthermore, Bu induced YAP expression and enhanced the translocation of YAP from the cytoplasm to the nucleus, which led to changes in the expression of mitogenesis genes, including TEAD1, TEAD4, CTGF and Cyr61. These results provide evidence that Bu stimulates the growth of human intestinal muscle cells by activation of YAP, which may be a potential treatment for improving intestinal adaptation. This article is protected by copyright. All rights reserved
      PubDate: 2017-10-11T02:00:52.442976-05:
      DOI: 10.1002/jcp.26149
  • Relationship of Human Herpes Virus 6 and Multiple Sclerosis: A Systematic
           Review and Meta-analysis
    • Authors: Ali Pormohammad; Taher Azimi, Fateme Falah, Ebrahim Faghihloo
      Abstract: BackgroundInfection with human herpes viruses has been suggested to contribute to multiple sclerosis (MS), while interaction between human herpes 6 (HHV6) and MS remain unclear yet. Here, we conducted a meta-analysis on the relationship of HHV6 infection and MS.Materials and MethodsAll related studies were collected from major databases. The analyses were performed by STATA 14 and Comprehensive Meta-Analysis V2.0 softwares. Pooled odds ratios (ORs) and 95% CIs were calculated from the raw data of the including studies by the random effects models when I2 >50% and fix model when I2 6 score that used serum/blood sample with OR of [6.7 (95% CI 4.8-8.6), P 
      PubDate: 2017-10-09T02:36:06.061244-05:
      DOI: 10.1002/jcp.26000
  • Heparan Sulfate Proteoglycan Deficiency Up-Regulates the Intracellular
           Production of Nitric Oxide in Chinese Hamster Ovary Cell Lines
    • Authors: Sheyla V. Lucena; Gioconda E. D. D. Moura, Tiago Rodrigues, Carolina M. Watashi, Fabiana H. Melo, Marcelo Y. Icimoto, Gustavo M. Viana, Helena B. Nader, Hugo P. Monteiro, Ivarne L. S. Tersariol, Fernando T. Ogata
      Abstract: We investigated the role of glycosaminoglycans (GAGs) in the regulation of endothelial nitric oxide synthase (eNOS) activity in wild-type CHO-K1 cells and in xylosyltransferase-deficient CHO-745 cells. GAGs inhibit the integrin/FAK/PI3K/AKT signaling pathway in CHO-K1 cells, decreasing the phosphorylation of eNOS at Ser1177. Furthermore, in CHO-K1 cells, eNOS and PKCα are localized at sphingolipid- and cholesterol-rich domains in the plasma membrane called caveolae. At caveolae, PKCα activation stimulates the phosphorylation of eNOS on Thr495, resulting in further inhibition of NO production in these cells. In our data, CHO-745 cells generate approximately 12-fold more NO than CHO-K1 cells. Increased NO production in CHO-745 cells promotes higher rates of protein S-nitrosylation and protein tyrosine nitration. Regarding reactive oxygen species (ROS) production, CHO-745 cells show lower basal levels of superoxide (O2-) than CHO-K1 cells. In addition, CHO-745 cells express higher levels of GPx, Trx1 and catalase than CHO-K1 cells, suggesting that CHO-745 cells are in a constitutive nitrosative/ oxidative stress condition. Accordingly, we showed that CHO-745 cells are more sensitive to oxidant-induced cell death than CHO-K1 cells. The high concentration of NO and reactive oxygen species generated by CHO-745 cells can induce simultaneous mitochondrial biogenesis and antioxidant gene expression. These observations led us to propose that GAGs are part of a regulatory mechanism that participates in eNOS activation and consequently regulates nitrosative/oxidative stress in CHO cells. This article is protected by copyright. All rights reserved
      PubDate: 2017-10-09T02:31:24.723196-05:
      DOI: 10.1002/jcp.26160
  • Transition metal dependent regulation of the signal transduction cascade
           driving oocyte meiosis
    • Authors: Stephanie Schaefer-Ramadan; Satanay Hubrack, Khaled Machaca
      Abstract: The G2-M transition of the cell cycle requires the activation of members of the Cdc25 dual specificity phosphatase family. Using Xenopus oocyte maturation as a model system, we have previously shown that chelation of transition metals blocks meiosis progression by inhibiting Cdc25C activation. Here, using approaches that allow for the isolation of very pure and active recombinant Cdc25C, we show that Cdc25C does not bind zinc as previously reported. Additionally, we show that mutants in the disordered C-terminal end of Cdc25C are poor initiators of meiosis, likely due to their inability to localize to the proper sub-cellular location. We further demonstrate that the transition metal chelator, TPEN, acts on or upstream of polo-like kinases in the oocyte to block meiosis progression. Together our results provide novel insights into Cdc25C structure-function relationship and the role of transition metals in regulating meiosis. This article is protected by copyright. All rights reserved
      PubDate: 2017-10-05T01:30:50.291403-05:
      DOI: 10.1002/jcp.26157
  • Blunting of estrogen modulation of cardiac cellular chymase/RAS activity
           and function in SHR
    • Authors: Sarfaraz Ahmad; Xuming Sun, Marina Lin, Jasmina Varagic, Gisele Zapata-Sudo, Carlos M. Ferrario, Leanne Groban, Hao Wang
      Abstract: The relatively low efficacy of ACE‐inhibitors in the treatment of heart failure in women after estrogen loss may be due to their inability to reach the intracellular sites at which angiotensin (Ang) II is generated and/or the existence of cell‐specific mechanisms in which ACE is not the essential processing pathway for Ang II formation. We compared the metabolic pathway for Ang II formation in freshly isolated myocytes (CMs) and non‐myocytes (NCMs) in cardiac membranes extracted from hearts of gonadal‐intact and ovariectomized (OVX) adult WKY and SHR rats. Plasma Ang II levels were higher in WKY vs. SHR (strain effect: WKY: 62 ± 6 pg/mL vs. SHR: 42 ± 9 pg/mL; P 
      PubDate: 2017-10-04T06:55:25.858439-05:
      DOI: 10.1002/jcp.26179
  • Comparison of genome‐wide analysis techniques to DNA methylation
           analysis in human cancer
    • Authors: Narges Soozangar; Mohammad R. Sadeghi, Farhad Jeddi, Mohammad H. Somi, Masoud Shirmohamadi, Nasser Samadi
      Abstract: DNA methylation was the first epigenetic modification to be detected in human cancers with specific relation to aberrant gene expression. Herein, DNA methylation analysis explains how epigenetic patterns affect gene expression level. Hypermethylation at tumor suppressor gene loci leads to increased tumorigenesis due to tumor suppressor gene silencing, whereas global hypomethylation of CpG islands (CGIs) is followed by genomic instability and aberrant activation of multiple oncogenes. Therefore, characterization of the genes which silenced or activated epigenetically in human tumor cells can improve our understanding of cancer biology. Different genome‐wide methodologies are applied to evaluate methylation status. Various commonly conducted techniques for this evaluation are reviewed in this paper. We provided comparative description of the procedures, advantages, and drawbacks of genome‐wide DNA methylation analysis methods and biological applications, to give information on selecting the appropriate method for different methylation studies. This article is protected by copyright. All rights reserved
      PubDate: 2017-10-04T06:51:17.572623-05:
      DOI: 10.1002/jcp.26176
  • Table of Contents, Editor's Choice, Highlights
    • First page: 1763
      PubDate: 2017-11-26T08:49:06.384887-05:
      DOI: 10.1002/jcp.26260
  • Troxerutin with copper generates oxidative stress in cancer cells: Its
           possible chemotherapeutic mechanism against hepatocellular carcinoma
    • Authors: Ariraman Subastri; Arumugam Suyavaran, Ezhuthupurakkal Preedia Babu, Subramaniyam Nithyananthan, Rajamani Barathidasan, Chinnasamy Thirunavukkarasu
      First page: 1775
      Abstract: Troxerutin (TXER) a rutin derivative is known for its anticancer effect against hepatocellular carcinoma (HCC). As part of large study, recently we have shown TXER interact with genetic material and its anti‐mutagenic property. In the present study we have explored its possible mode of action in HCC. Since TXER alone did not show significant anticancer effect on Huh‐7 cells, in vitro biochemical assays were performed for determining anticancer efficacy of TXER + metal complex using transition metals such as Cu, Zn, and Fe. The anticancer efficacy of TXER + Cu on Huh‐7 cells were evaluated using MTT assay, DCFDA, JC‐1 staining, comet assay, cell cycle analysis, immunocytochemistry, and Western blotting. Non‐toxic nature of TXER was analyzed on primary rat hepatocytes. The in vivo efficacy of TXER was tested in N‐nitrosodiethylamine initiated and γ‐benzene hexachloride and partial hepatectomy promoted rat liver cancer. Liver markers, transition metal levels, histopathological examination, and expression levels of GST‐P, 8‐OHdG and Ki‐67 were studied to assess the in vivo anticancer effect of TXER. We observed that TXER + Cu induced extensive cellular death on Huh‐7 cells through generating free radicals and did not possess any toxic effect on normal hepatocytes. The in vivo studies revealed that TXER possess significant anti‐cancer effect as assessed through improved liver markers and suppressed GST‐P, 8‐OHdG, and Ki‐67 expression. TXER treatment reduced the hepatic Cu level in cancer bearing animals. Current study brings the putative mechanism involved in anti‐cancer effect of TXER, further it will help to formulate phytoconstituents coupled anti‐cancer drug for effective treatment of HCC.Cu is known to be elevated in cancer cells. Drug may have the possibility to interact with Cu and to alter homeostasis of cancer cells. In vitro study, shows that TXER + Cu complex exert significant Huh‐7 liver cancer cell death through apoptosis. Anticancer effect of TXER was confirmed in rat in vivo model. Our findings revealed the putative mechanisms behind the anticancer effect of TXER in hepatocellular carcinoma.
      PubDate: 2017-08-03T06:21:00.325178-05:
      DOI: 10.1002/jcp.26061
  • Evaluation of BAG3 levels in healthy subjects, hypertensive patients, and
           hypertensive diabetic patients
    • Authors: Giuseppe Derosa; Pamela Maffioli, Alessandra Rosati, De Marco M, Anna Basile, Angela D'Angelo, Davide Romano, Amirhossein Sahebkar, Antonia Falco, Maria C. Turco
      First page: 1791
      Abstract: BAG3 is a member of human BAG (Bcl‐2‐associated athanogene) proteins and plays a role in apoptosis, cell adhesion, cytoskeleton remodeling, and autophagy. The aim of this study was to evaluate BAG3 levels in healthy subjects, hypertensive patients, and hypertensive diabetic patients. We enrolled 209 Caucasian adults, of both sex, 18–75 years of age, 77 were healthy controls, 62 were affected by hypertension, and 70 were affected by hypertension and type 2 diabetes. All patients underwent an assessment that included medical history, physical examination, vital signs, a 12‐lead electrocardiogram, measurements of systolic (SBP), and diastolic blood pressure (DBP), heart rate (HR), fasting plasma glucose (FPG), glycated hemoglobin (HbA1c), triglycerides (TG), transaminases, high sensitivity C‐reactive protein (Hs‐CRP), and BAG3. We observed higher blood pressure values in hypertensive, and hypertensive diabetic patients compared to controls. As expected, FPG and HbA1c were higher in diabetic hypertensive patients, compared to the other two groups. No Tg levels differences were recorded among the three groups. Hs‐CRP was higher in diabetic hypertensive patients compared to healthy subjects. Finally, BAG3 levels were higher in hypertensives, and hypertensive diabetic patients compared to controls. We observed higher levels of BAG3 in hypertensive patients compared to healthy controls, and even higher levels in hypertensive diabetic patients compared to healthy subjects. This paper could be the first of a long way to identify potential involvement of deregulated BAG3 levels in cardiometabolic diseases.BAG3 is a member of human BAG (Bcl‐2‐associated athanogene) proteins and plays a role in apoptosis, cell adhesion, cytoskeleton remodeling, and autophagy.
      PubDate: 2017-08-25T05:45:22.400531-05:
      DOI: 10.1002/jcp.26093
  • Phosphatidylinositol 3‐kinase inhibition potentiates glucocorticoid
           response in B‐cell acute lymphoblastic leukemia
    • Authors: Cecilia Evangelisti; Alessandra Cappellini, Mariana Oliveira, Rita Fragoso, João T. Barata, Alice Bertaina, Franco Locatelli, Carolina Simioni, Luca M. Neri, Francesca Chiarini, Annalisa Lonetti, Francesca Buontempo, Ester Orsini, Andrea Pession, Lucia Manzoli, Alberto Maria Martelli, Camilla Evangelisti
      First page: 1796
      Abstract: Despite remarkable progress in polychemotherapy protocols, pediatric B‐cell acute lymphoblastic leukemia (B‐ALL) remains fatal in around 20% of cases. Hence, novel targeted therapies are needed for patients with poor prognosis. Glucocorticoids (GCs) are drugs commonly administrated for B‐ALL treatment. Activation of the phosphatidylinositol 3‐kinase (PI3K)/Akt/mammalian target of rapamycin signaling pathway is frequently observed in B‐ALL and contributes to GC‐resistance. Here, we analyzed for the first time to our knowledge, the therapeutic potential of pan and isoform‐selective PI3K p110 inhibitors, alone or combined with dexamethasone (DEX), in B‐ALL leukemia cell lines and patient samples. We found that a pan PI3K p110 inhibitor displayed the most powerful cytotoxic effects in B‐ALL cells, by inducing cell cycle arrest and apoptosis. Both a pan PI3K p110 inhibitor and a dual γ/δ PI3K p110 inhibitor sensitized B‐ALL cells to DEX by restoring nuclear translocation of the GC receptor and counteracted stroma‐induced DEX‐resistance. Finally, gene expression analysis documented that, on one hand the combination consisting of a pan PI3K p110 inhibitor and DEX strengthened the DEX‐induced up‐ or down‐regulation of several genes involved in apoptosis, while on the other, it rescued the effects of genes that might be involved in GC‐resistance. Overall, our findings strongly suggest that PI3K p110 inhibition could be a promising strategy for treating B‐ALL patients by improving GC therapeutic effects and/or overcoming GC‐resistance.We found that a pan PI3K p110 inhibitor displayed the most powerful cytotoxic effects in B‐ALL cells, by inducing cell cycle arrest and apoptosis. Both a pan PI3K p110 inhibitor and a dual γ/δ PI3K p110 inhibitor sensitized B‐ALL cells to DEX by restoring nuclear translocation of the GC receptor and counteracted stroma‐induced DEX‐resistance. Overall, our findings strongly suggest that PI3K p110 inhibition could be a promising strategy for treating B‐ALL patients by improving GC therapeutic effects and/or overcoming GC‐resistance.
      PubDate: 2017-09-07T04:40:54.077509-05:
      DOI: 10.1002/jcp.26135
  • Combined effects of bone morphogenetic protein 10 and crossveinless‐2 on
           cardiomyocyte differentiation in mouse adipocyte‐derived stem cells
    • Authors: Medet Jumabay; Jiayinaguli Zhumabai, Nurlan Mansurov, Katharine C. Niklason, Pierre J. Guihard, Mark R. Cubberly, Alan M. Fogelman, Luisa Iruela-Arispe, Yucheng Yao, Arman Saparov, Kristina I. Boström
      First page: 1812
      Abstract: Bone morphogenetic protein (BMP) 10, a cardiac‐restricted BMP family member, is essential in cardiomyogenesis, especially during trabeculation. Crossveinless‐2 (CV2, also known as BMP endothelial cell precursor derived regulator [BMPER]) is a BMP‐binding protein that modulates the activity of several BMPs. The objective of this study was to examine the combined effects of BMP10 and CV2 on cardiomyocyte differentiation using mouse dedifferentiated fat (mDFAT) cells, which spontaneously differentiate into cardiomyocyte‐like cells, as a model. Our results revealed that CV2 binds directly to BMP10, as determined by co‐immunoprecipitation, and inhibits BMP10 from initiating SMAD signaling, as determined by luciferase reporter gene assays. BMP10 treatment induced mDFAT cell proliferation, whereas CV2 modulated the BMP10‐induced proliferation. Differentiation of cardiomyocyte‐like cells proceeded in a reproducible fashion in mDFAT cells, starting with small round Nkx2.5‐positive progenitor cells that progressively formed myotubes of increasing length that assembled into beating colonies and stained strongly for Troponin I and sarcomeric alpha‐actinin. BMP10 enhanced proliferation of the small progenitor cells, thereby securing sufficient numbers to support formation of myotubes. CV2, on the other hand, enhanced formation and maturation of large myotubes and myotube‐colonies and was expressed by endothelial‐like cells in the mDFAT cultures. Thus BMP10 and CV2 have important roles in coordinating cardiomyogenesis in progenitor cells.BMP10 and CV2 affect cardiomyogenesis in vitro.
      PubDate: 2017-06-06T07:15:37.510972-05:
      DOI: 10.1002/jcp.25983
  • A perspective on stem cell therapy for ear disorders
    • Authors: Rahul Mittal; Hyunseo D. Jung, Jeenu Mittal, Adrien A. Eshraghi
      First page: 1823
      Abstract: The use of stem cells in cell‐based therapy is an emerging concept for the treatment of ear disorders. Tympanic membrane perforation (TMP) and inner ear disorders are some of the most commonly presented otologic disorders that can benefit from advances in cell‐based therapy. Studies have already demonstrated that stem cell‐based therapy can potentially be an effective treatment modality for acute and chronic TMP. Recent studies have also shown promise in application of cell‐based approach to treat inner ear dysfunction. In this perspective, we will discuss the recent advancements regarding the use of cell‐based therapy for ear disorders.
      PubDate: 2017-08-03T06:10:25.117767-05:
      DOI: 10.1002/jcp.26048
  • Stem cells applications in bone and tooth repair and regeneration: New
           insights, tools, and hopes
    • Authors: Eiman Abdel Meguid; Yuehai Ke, Junfeng Ji, Ahmed H.K. El-Hashash
      First page: 1825
      Abstract: The exploration of stem and progenitor cells holds promise for advancing our understanding of the biology of tissue repair and regeneration mechanisms after injury. This will also help in the future use of stem cell therapy for the development of regenerative medicine approaches for the treatment of different tissue‐species defects or disorders such as bone, cartilages, and tooth defects or disorders. Bone is a specialized connective tissue, with mineralized extracellular components that provide bones with both strength and rigidity, and thus enable bones to function in body mechanical supports and necessary locomotion process. New insights have been added to the use of different types of stem cells in bone and tooth defects over the last few years. In this concise review, we briefly describe bone structure as well as summarize recent research progress and accumulated information regarding the osteogenic differentiation of stem cells, as well as stem cell contributions to bone repair/regeneration, bone defects or disorders, and both restoration and regeneration of bones and cartilages. We also discuss advances in the osteogenic differentiation and bone regeneration of dental and periodontal stem cells as well as in stem cell contributions to dentine regeneration and tooth engineering.Stem cell applications have great potentials and hopes for both bone and tooth repair and regeneration.
      PubDate: 2017-05-23T11:55:27.669995-05:
      DOI: 10.1002/jcp.25940
  • Role of adenosine signaling in the pathogenesis of breast cancer
    • Authors: Amirhossein Bahreyni; Seyed Sattar Samani, Farzad Rahmani, Reihane Behnam-Rassouli, Majid Khazaei, Mikhail Ryzhikov, Mohammad Reza Parizadeh, Amir Avan, Seyed Mahdi Hassanian
      First page: 1836
      Abstract: The plasma level of adenosine increases under ischemic and inflamed conditions in tumor microenvironment. Adenosine elicits a range of signaling pathways in tumors, resulting in either inhibition or enhancement of tumor growth depending upon different subtypes of adenosine receptors activation and type of cancer. Metabolism of adenosine‐5′‐triphosphate (ATP) and its derivatives including adenosine is dysregulated in the breast tumor microenvironment, supporting the role of this metabolite in the pathogenesis of breast cancer. Adenosine regulates inflammation, apoptosis, cell proliferation, and metastasis in breast cancer cells. This review summarizes the role of adenosine in the pathogenesis of breast cancer for a better understanding and hence a better management of this disease.Metabolism of adenosine‐5′‐triphosphate (ATP) and its derivatives including adenosine is dysregulated in the breast tumor microenvironment, supporting the role of this metabolite in the pathogenesis of breast cancer. Adenosine regulates inflammation, apoptosis, cell proliferation, and metastasis in breast cancer cells. This review summarizes the role of adenosine in the pathogenesis of breast cancer for a better understanding and hence a better management of this disease.
      PubDate: 2017-05-23T11:25:25.638115-05:
      DOI: 10.1002/jcp.25944
  • CRISPR/Cas9: An RNA‐guided highly precise synthetic tool for plant
           genome editing
    • Authors: Yeliz Demirci; Baohong Zhang, Turgay Unver
      First page: 1844
      Abstract: CRISPR/Cas9 is a newly developed and naturally occurred genome editing tool, which is originally used by bacteria for immune defence. In the past years, it has been quickly employed and modified to precisely edit genome sequences in both plants and animals. Compared with the well‐developed zinc finger nucleases (ZFNs) and transcription activator‐like effector nucleases (TALENs), CRISPR/Cas9 has lots of advantages, including easier to design and implement, higher targeting efficiency, and less expensive. Thus, it is becoming one of the most powerful tools for knockout of an individual gene as well as insertion of one gene and/or control of gene transcription. Studies have shown that CRISPR/Cas9 is a great tool to edit many genes in a variety of plant species, including the model plant species as well as agriculturally important crops, such as cotton, maize, wheat, and rice. CRISPR/Cas9‐based genome editing can be used for plant functional studies and plant improvement to yield, quality, and tolerance to environmental stress.CRISPR/Cas9 is a newly developed and naturally occurred genome editing tool, which is originally used by bacteria for immune defence. In the past years, it has been quickly employed and modified to precisely edit genome sequences in both plants and animals, including the model plant species as well as agriculturally important crops, such as cotton, maize, and rice. Compared with the well‐developed zinc finger nucleases (ZFNs) and transcription activator‐like effector nucleases (TALENs), CRISPR/Cas9 has lots of advantages, including easier to design and implement, higher targeting efficiency, and less expensive.
      PubDate: 2017-06-06T06:45:50.01835-05:0
      DOI: 10.1002/jcp.25970
  • Effects of electrical stimulation on cell proliferation and apoptosis
    • Authors: Maria R. Love; Siripong Palee, Siriporn C. Chattipakorn, Nipon Chattipakorn
      First page: 1860
      Abstract: The application of exogenous electrical stimulation (ES) to cells in order to manipulate cell apoptosis and proliferation has been widely investigated as a possible method of treatment in a number of diseases. Alteration of the transmembrane potential of cells via ES can affect various intracellular signaling pathways which are involved in the regulation of cellular function. Controversially, several types of ES have proved to be effective in both inhibiting or inducing apoptosis, as well as increasing proliferation. However, the mechanisms through which ES achieves this remain fairly unclear. The aim of this review was to comprehensively summarize current findings from in vitro and in vivo studies on the effects of different types of ES on cell apoptosis and proliferation, highlighting the possible mechanisms through which ES induced these effects and define the optimum parameters at which ES can be used. Through this we hope to provide a greater insight into how future studies can most effectively use ES at the clinical trial stage.Roles of electrical stimulation on cells to manipulate cell apoptosis and proliferation has been widely investigated. This review summarizes the reports of various types of electrical stimulation on these effects.
      PubDate: 2017-06-06T06:56:23.82224-05:0
      DOI: 10.1002/jcp.25975
  • Biological function and histone recognition of family IV
           bromodomain‐containing proteins
    • Authors: Jonathan T. Lloyd; Karen C. Glass
      First page: 1877
      Abstract: Bromodomain proteins function as epigenetic readers that recognize acetylated histone tails to facilitate the transcription of target genes. There are approximately 60 known human bromodomains, which are divided into eight sub‐families based on structural conservation. The bromodomain‐containing proteins in family IV include seven members (BRPF1, BRPF2, BRPF3, BRD7, BRD9, ATAD2, and ATAD2b). The bromodomains of each of these proteins recognize and bind acetyllysine residues on histone tails protruding from the nucleosome. However, the histone marks recognized by each bromodomain protein can be very different. The BRPF1 subunit of the MOZ histone acetyltransferase (HAT) recognizes acetylated histones H2AK5ac, H4K12ac, H3K14ac, H4K8ac, and H4K5ac. While the bromodomain of BRD7, a member of the SWI/SNF complex, was shown to preferentially recognize acetylated histones H3K9ac, H3K14ac, H4K8ac, H4K12ac, and H4K16ac. The bromodomains of BRPF2 and BRPF3 have similar sequences, and function as part of the HBO1 HAT complex, but there is limited data on which histone ligands they bind. Similarly, there is little known about the histone targets of the BRD9 and ATAD2b bromodomain proteins. Interestingly, the ATAD2 bromodomain was recently shown to preferentially bind to the di‐acetylated H4K5acK12ac mark found in newly synthesized histones following DNA replication. However, despite the physiological importance of the family IV bromodomains, little is known about how they function at the molecular or atomic level. In this review, we summarize our understanding of how family IV bromodomains recognize and select for acetyllysine marks and discuss the importance of acetylated histone recognition for their biological functions.The bromodomain‐containing proteins in family IV include seven members (BRPF1, BRPF2, BRPF3, BRD7, BRD9, ATAD2, and ATAD2b), each of which recognize and bind acetyllysine residues on histone tails. However, the histone marks recognized by each bromodomain protein can be very different. Here, we summarize our understanding of how family IV bromodomains recognize and select for acetyllysine marks and discuss the importance of acetylated histone recognition for their biological functions.
      PubDate: 2017-06-13T04:50:29.898432-05:
      DOI: 10.1002/jcp.26010
  • The emerging role of NPNT in tissue injury repair and bone homeostasis
    • Authors: Youqiang Sun; Vincent Kuek, Heng Qiu, Jennifer Tickner, Leilei Chen, Haibin Wang, Wei He, Jiake Xu
      First page: 1887
      Abstract: Nephronectin (NPNT), a highly conserved extracellular matrix protein, plays an important role in regulating cell adhesion, differentiation, spreading, and survival. NPNT protein belongs to the epidermal growth factor (EGF)‐like superfamily and exhibits several common structural determinants; including EGF‐like repeat domains, MAM domain (Meprin, A5 Protein, and Receptor Protein‐Tyrosine Phosphatase µ), RGD motif (Arg‐Gly‐Asp) and a coiled‐coil domain. It regulates integrins‐mediated signaling pathways via the interaction of its RGD motif with integrin α8β1. Recent studies revealed that NPNT is involved in kidney development, renal injury repair, atrioventricular canal differentiation, pulmonary function, and muscle cell niche maintenance. Moreover, NPNT regulates osteoblast differentiation and mineralization, as well as osteogenic angiogenesis. Altered expression of NPNT has been linked with the progression of certain types of cancers, such as spontaneous breast tumor metastasis and malignant melanoma. Interestingly, NPNT gene expression can be regulated by a range of external factors such as tumor necrosis factor alpha (TNF‐α), transforming growth factor beta (TGF‐β), oncostatin M (OSM), bone morphogenic protein 2 (BMP2), Wnt3a, Vitamin D3, and microRNA‐378 (miR378). Further understanding the cellular and molecular mechanisms by which NPNT regulates tissue homeostasis in an organ‐specific manner is critical in exploring NPNT as a therapeutic target for tissue regeneration and tissue engineering.Recently studies show that NPNT regulates osteoblast differentiation and mineralization, as well as osteogenic angiogenesis. Interestingly, NPNT gene expression can be regulated by a range of external factors such as tumor necrosis factor alpha (TNF‐α), transforming growth factor beta (TGF‐β), oncostatin M (OSM), bone morphogenic protein 2 (BMP2), Wnt3a, Vitamin D3 and microRNA‐378 (miR378). Further understanding the cellular and molecular mechanisms by which NPNT regulates tissue homeostasis in an organ‐specific manner is critical in exploring NPNT as a therapeutic target for tissue regeneration and tissue engineering.
      PubDate: 2017-06-16T07:10:31.789611-05:
      DOI: 10.1002/jcp.26013
  • NAD+: A big player in cardiac and skeletal muscle remodeling and aging
    • Authors: Pankaj Chaturvedi; Suresh C. Tyagi
      First page: 1895
      Abstract: In the past decade, NAD+ has gained importance for its beneficial effects as antioxidant and anti‐aging molecule. A paper in science by Zhang et al. () has described that NAD+ when replenished, ameliorates muscle dystrophy in mice by improving mitochondrial function. NAD+ was also demonstrated by the authors to improve the life span of mice. Cox et al. () demonstrated the cardiac effects of NAD+ which mitigated chronic heart failure via mitochondrial redox state mechanism. Cox et al. () also demonstrated that NAD+ is provided in the drinking water, it improves cardiac relaxation in volume overload model of heart failure. Although NAD+ has a profound anti‐aging and anti‐oxidant effects, its effect on humans and use as a dietary supplement needs more exploration.NAD+ has gained importance for its beneficial effects as antioxidant and antiaging molecule, NAD+ when replenished, ameliorates muscle dystrophy in mice by improving mitochondrial function.
      PubDate: 2017-07-07T10:10:24.513005-05:
      DOI: 10.1002/jcp.26014
  • Intimate connections: Inositol pyrophosphates at the interface of
           metabolic regulation and cell signaling
    • Authors: Stephen B. Shears
      First page: 1897
      Abstract: Inositol pyrophosphates are small, diffusible signaling molecules that possess the most concentrated three‐dimensional array of phosphate groups in Nature; up to eight phosphates are crammed around a six‐carbon inositol ring. This review discusses the physico‐chemical properties of these unique molecules, and their mechanisms of action. Also provided is information on the enzymes that regulate the levels and hence the signaling properties of these molecules. This review pursues the idea that many of the biological effects of inositol pyrophosphates can be rationalized by their actions at the interface of cell signaling and metabolism that is essential to cellular and organismal homeostasis.A review of inositol pyrophosphate metabolism and their actions at the interface of metabolic control and cell signaling.
      PubDate: 2017-06-15T04:35:24.199478-05:
      DOI: 10.1002/jcp.26017
  • Effects of mechanical loading on human mesenchymal stem cells for
           cartilage tissue engineering
    • Authors: Jane Ru Choi; Kar Wey Yong, Jean Yu Choi
      First page: 1913
      Abstract: Today, articular cartilage damage is a major health problem, affecting people of all ages. The existing conventional articular cartilage repair techniques, such as autologous chondrocyte implantation (ACI), microfracture, and mosaicplasty, have many shortcomings which negatively affect their clinical outcomes. Therefore, it is essential to develop an alternative and efficient articular repair technique that can address those shortcomings. Cartilage tissue engineering, which aims to create a tissue‐engineered cartilage derived from human mesenchymal stem cells (MSCs), shows great promise for improving articular cartilage defect therapy. However, the use of tissue‐engineered cartilage for the clinical therapy of articular cartilage defect still remains challenging. Despite the importance of mechanical loading to create a functional cartilage has been well demonstrated, the specific type of mechanical loading and its optimal loading regime is still under investigation. This review summarizes the most recent advances in the effects of mechanical loading on human MSCs. First, the existing conventional articular repair techniques and their shortcomings are highlighted. The important parameters for the evaluation of the tissue‐engineered cartilage, including chondrogenic and hypertrophic differentiation of human MSCs are briefly discussed. The influence of mechanical loading on human MSCs is subsequently reviewed and the possible mechanotransduction signaling is highlighted. The development of non‐hypertrophic chondrogenesis in response to the changing mechanical microenvironment will aid in the establishment of a tissue‐engineered cartilage for efficient articular cartilage repair.The conventional articular cartilage repair techniques, such as ACI, microfracture, and mosaicplasty, have many shortcomings which negatively affect their clinical outcomes. Cartilage tissue engineering shows great promise for improving articular cartilage defect therapy. Mechanical loading is a key cartilage engineering approach, which could induce non‐hypertrophic chondrogenesis of human MSCs to produce a fully functional tissue‐engineered cartilage for articular cartilage defect therapy.
      PubDate: 2017-06-12T05:25:38.577019-05:
      DOI: 10.1002/jcp.26018
  • Genetic susceptibility in cervical cancer: From bench to bedside
    • Authors: Afsane Bahrami; Malihe Hasanzadeh, Soodabeh Shahidsales, Marjaneh Farazestanian, Seyed Mahdi Hassanian, Mehrdad Moetamani Ahmadi, Mina Maftouh, Masoumeh Gharib, Zohreh Yousefi, Sima Kadkhodayan, Gordon A. Ferns, Amir Avan
      First page: 1929
      Abstract: Cervical cancer (CC) is the third most common malignancy in women globally, and persistent infection with the oncogenic human papillomaviruses (HPV) is recognized as the major risk factor. The pathogenesis of CC relies on the interplay between the tumorigenic properties of the HPV and host factors. Host‐related genetic factors, including the presence of susceptibility loci for cervix tumor is substantial importance. Preclinical and genome‐wide association studies (GWAS) have reported the associations of genetic variations in several susceptibility loci for the development of cervical cancer. However, many of these reports are inconsistent. In this review, we discuss the findings to date of candidate gene association studies, and GWAS in cervical cancer. The associations between these genetic variations with response to chemotherapy are also discussed.
      PubDate: 2017-09-07T04:41:02.857652-05:
      DOI: 10.1002/jcp.26019
  • Co‐culture systems‐based strategies for articular cartilage
           tissue engineering
    • Authors: Yu Zhang; Weimin Guo, Mingjie Wang, Chunxiang Hao, Liang Lu, Shuang Gao, Xueliang Zhang, Xu Li, Mingxue Chen, Penghao Li, Peng Jiang, Shibi Lu, Shuyun Liu, Quanyi Guo
      First page: 1940
      Abstract: Cartilage engineering facilitates repair and regeneration of damaged cartilage using engineered tissue that restores the functional properties of the impaired joint. The seed cells used most frequently in tissue engineering, are chondrocytes and mesenchymal stem cells. Seed cells activity plays a key role in the regeneration of functional cartilage tissue. However, seed cells undergo undesirable changes after in vitro processing procedures, such as degeneration of cartilage cells and induced hypertrophy of mesenchymal stem cells, which hinder cartilage tissue engineering. Compared to monoculture, which does not mimic the in vivo cellular environment, co‐culture technology provides a more realistic microenvironment in terms of various physical, chemical, and biological factors. Co‐culture technology is used in cartilage tissue engineering to overcome obstacles related to the degeneration of seed cells, and shows promise for cartilage regeneration and repair. In this review, we focus first on existing co‐culture systems for cartilage tissue engineering and related fields, and discuss the conditions and mechanisms thereof. This is followed by methods for optimizing seed cell co‐culture conditions to generate functional neo‐cartilage tissue, which will lead to a new era in cartilage tissue engineering.Articular cartilage cells have unique growth environment, including hypoxia, mechnical stimulation, and massive extracellular matrix. Co‐culture system can provide cartilage cells biomimetic microenvironment to construct high quality transplant to repair injurious cartilage. In this review, we give a conclusion of co‐culture systems of articular cartilage tissue engineering and find a way to improve co‐culture system to promote repair and regeneration of articular cartilage.
      PubDate: 2017-09-08T03:30:30.838858-05:
      DOI: 10.1002/jcp.26020
  • Gingival spheroids possess multilineage differentiation potential
    • Authors: Rajasekaran Subbarayan; Dinesh Murugan Girija, Suresh Ranga Rao
      First page: 1952
      Abstract: Recently studies have demonstrated HGMSCs as ideal candidates for regenerative study. Interestingly we found that HGMSCs derived spheroids are more potent and maintain the properties of stemness convincingly compared to conventional culture methods. During the culture, GMSCs instinctively accumulated into spheroids and display multipotent STRO‐1 and Vimentin‐positive cells. Reduced phenotypic expression of CD73, CD105, and elevated expression STRO‐1 and CD‐34. Pluripotent nature of S‐GMSCs putatively shown the expression of OCT4A, NANOG, SOX‐2, SSEA4, TRA‐1‐60, and TRA‐181. Also, levels of protein are much higher in spheroid than dissociated culture. On endothelial induction, spheroid differentiated and developed a vascular structure with positive expression of CD31 and on neuronal induction showed positivity for TUJ1 and E‐Cadherin. Importantly, undifferentiated state of S‐GMSCs exhibited significant upregulation of aforementioned pluripotent genes and lack of pro‐inflammatory cytokines IL‐6 and amplified ARF signal confirming that the spheroids are not teratoma formation. However, higher of CAP1, CP, TGFβ, OPN, PPARɣ, TUJ1, and NESTIN expression observed in spheroids, and minimal expression of the same markers were observed in adherent GMSCs respectively. Ahead of dissociated gingival culture, spheroid provides enhanced viable, pluripotent, and multilineage ability. This study suggested that S‐GMSCs increased the chances of therapeutic efficacy in the regenerative applications.This study suggested that Spheroid‐GMSCs increased the chances of therapeutic efficacy in the regenerative applications.
      PubDate: 2017-06-05T08:15:29.041275-05:
      DOI: 10.1002/jcp.25894
  • Three‐dimensional culture of buffalo granulosa cells in hanging drop
           mimics the preovulatory follicle stage
    • Authors: Monica Yadav; Himanshu Agrawal, Mamta Pandey, Dheer Singh, Suneel K. Onteru
      First page: 1959
      Abstract: Granulosa cell (GC) culture models mimicking the intrafollicular environment are limited. Such models have a great potential in reproductive toxicity studies. The buffalo, a monovulatory species like humans, could be a better model than polyovulatory rodents. Therefore, we targeted the development and characterization of three‐dimensional (3D) culture systems for buffalo GCs. The GCs from small ovarian follicles (SF) maintained the CYP19 gene expression for 144 hr in a 2D culture system. Hence, GCs from SF were cultured directly in 3D using hanging drop and Poly‐([2‐hydroxyethyl methacrylate]) (polyHEMA) methods in the DMEM media containing 1 ng/ml FSH and 10 ng/ml IGF‐1 for 144 hr. The expression profile of nine GC‐specific transcripts; CYP19, TNFAIP6, AMH, PTI, NR4A1, FSHR, RUNX, LHR, and COX2/PTGS2; revealed that 3D‐spheroids developed in hanging drop method maintained the GC phenotype of preovulatory follicles. Therefore, hanging drop method is a best method for culturing GCs to mimic the intrafollicular environment.Buffalo granulosa cells formed 3D‐spheroids in hanging drop method, which maintained their phenotype as of preovulatory follicles. Therefore, hanging drop method is the best method for culturing GCs to mimic the intrafollicular environment.
      PubDate: 2017-05-03T09:10:50.119792-05:
      DOI: 10.1002/jcp.25909
  • A highly efficient method for extracting next‐generation sequencing
           quality RNA from adipose tissue of recalcitrant animal species
    • Authors: Davinder Sharma; Naresh Golla, Dheer Singh, Suneel K. Onteru
      First page: 1971
      Abstract: The next‐generation sequencing (NGS) based RNA sequencing (RNA‐Seq) and transcriptome profiling offers an opportunity to unveil complex biological processes. Successful RNA‐Seq and transcriptome profiling requires a large amount of high‐quality RNA. However, NGS‐quality RNA isolation is extremely difficult from recalcitrant adipose tissue (AT) with high lipid content and low cell numbers. Further, the amount and biochemical composition of AT lipid varies depending upon the animal species which can pose different degree of resistance to RNA extraction. Currently available approaches may work effectively in one species but can be almost unproductive in another species. Herein, we report a two step protocol for the extraction of NGS quality RNA from AT across a broad range of animal species.NGS‐quality RNA isolation is extremely difficult from recalcitrant adipose tissue (AT) with high lipid content and low cell numbers. Currently available approaches yield variable and unsatisfactory results with AT samples.A highly efficient novel protocol was developed for the extraction of NGS quality RNA from AT across a broad range of animal species.
      PubDate: 2017-06-05T08:30:20.425517-05:
      DOI: 10.1002/jcp.25951
  • Effect of stress on human biology: Epigenetics, adaptation, inheritance,
           and social significance
    • Authors: David T. Denhardt
      First page: 1975
      Abstract: I present a brief introduction to epigenetics, focused primarily on methylation of the genome and various regulatory RNAs, modifications of associated histones, and their importance in enabling us to adapt to real and changing environmental, developmental, and social circumstances. Following this is a more extensive overview of how it impacts our inheritance, our entire life (which changes as we age), and how we interact with others. Throughout, I emphasize the critical influence that stress, of many varieties exerts, via epigenetic means, on much of how we live and survive, mostly in the brain. I end with a short section on multigenerational transmission, drugs, and the importance of both social life and early life experiences in the development of adult diseases. There will be nothing about cancer. Although epigenetics is critical in that field, it is a whole different cobweb of complications (some involving stress).This review focuses on the impact of stress of many kinds on human physiology. It discusses DNA and histone modifications and regulation by various forms of RNA. It ends with a critical analysis of the impact of drugs and social interactions on the individual and transgenerational inheritance.
      PubDate: 2017-04-27T18:56:41.592105-05:
      DOI: 10.1002/jcp.25837
  • Mimicking exercise in three‐dimensional bioengineered skeletal muscle to
    • Authors: Andreas M. Kasper; Daniel C. Turner, Neil R. W. Martin, Adam P. Sharples
      First page: 1985
      Abstract: Bioengineering of skeletal muscle in vitro in order to produce highly aligned myofibres in relevant three dimensional (3D) matrices have allowed scientists to model the in vivo skeletal muscle niche. This review discusses essential experimental considerations for developing bioengineered muscle in order to investigate exercise mimicking stimuli. We identify current knowledge for the use of electrical stimulation and co‐culture with motor neurons to enhance skeletal muscle maturation and contractile function in bioengineered systems in vitro. Importantly, we provide a current opinion on the use of acute and chronic exercise mimicking stimuli (electrical stimulation and mechanical overload) and the subsequent mechanisms underlying physiological adaptation in 3D bioengineered muscle. We also identify that future studies using the latest bioreactor technology, providing simultaneous electrical and mechanical loading and flow perfusion in vitro, may provide the basis for advancing knowledge in the future. We also envisage, that more studies using genetic, pharmacological, and hormonal modifications applied in human 3D bioengineered skeletal muscle may allow for an enhanced discovery of the in‐depth mechanisms underlying the response to exercise in relevant human testing systems. Finally, 3D bioengineered skeletal muscle may provide an opportunity to be used as a pre‐clinical in vitro test‐bed to investigate the mechanisms underlying catabolic disease, while modelling disease itself via the use of cells derived from human patients without exposing animals or humans (in phase I trials) to the side effects of potential therapies.This review discusses the current understanding, advances, and future directions for the stimulation of three‐dimensional bioengineered skeletal muscle to investigate the mechanisms of physiological adaptation to exercise.
      PubDate: 2017-03-21T08:45:27.729577-05:
      DOI: 10.1002/jcp.25840
  • Molecular pathogenesis in chronic obstructive pulmonary disease and
           therapeutic potential by targeting AMP‐activated protein kinase
    • Authors: Zhihui Zhang; Xiaoyu Cheng, Li Yue, Wenhui Cui, Wencheng Zhou, Jian Gao, Hongwei Yao
      First page: 1999
      Abstract: Chronic obstructive pulmonary disease (COPD) is the third leading cause of death worldwide, which is characterized by a persistent airflow limitation caused by chronic inflammatory responses to noxious particles or gases. Cigarette smoke and environmental pollutions are major etiological factors for causing COPD. It has been shown that cigarette smoking causes abnormal inflammatory responses, cellular senescence, mitochondrial dysfunction and metabolic dysregulation, suggesting their involvement in the development of COPD. Although the medical care and treatment have advanced, there are no effective therapies to stop or reverse lung destruction in COPD/emphysema. AMP‐activated protein kinase (AMPK) is a serine threonine kinase with α, β, and γ subunits that are highly conserved through evolution. AMPK has been shown to regulate bioenergetics, inflammatory responses, senescence, and metabolism. This review focused on the updated understanding of molecular pathogenesis of COPD, and highlighted the crucial roles of AMPK in lung abnormalities as well as discussed the potential therapeutics of AMPK activators in preventing and halting the progression of COPD.Cigarette smoke as well as other gases and particles cause inflammatory responses, senescence, mitochondrial dysfunction, and metabolic dysregulation, leading to lung destruction and subsequently COPD/emphysema. AMPK is reduced by cigarette smoke and in lungs of COPD patients, and AMPK activators protect against aforementioned cellular processes and pulmonary emphysema.
      PubDate: 2017-04-27T19:09:53.097059-05:
      DOI: 10.1002/jcp.25844
  • Interplay between miRNAs and human diseases
    • Authors: Prosenjit Paul; Anindya Chakraborty, Debasree Sarkar, Malobika Langthasa, Musfhia Rahman, Minakshi Bari, RK. Sanamacha Singha, Arup Kumar Malakar, Supriyo Chakraborty
      First page: 2007
      Abstract: MicroRNAs (miRNAs) are endogenous, non‐coding RNAs, which have evoked a great deal of interest due to their importance in many aspects of homeostasis and diseases. MicroRNAs are stable and are essential components of gene regulatory networks. They play a crucial role in healthy individuals and their dysregulations have also been implicated in a wide range of diseases, including diabetes, cardiovascular disease, kidney disease, and cancer. This review summarized the current understanding of interactions between miRNAs and different diseases and their role in disease diagnosis and therapy.This review summarized the current understanding of interactions between miRNAs and different diseases and their role in disease diagnosis and therapy.
      PubDate: 2017-04-27T18:40:24.670527-05:
      DOI: 10.1002/jcp.25854
  • Modulating tumor hypoxia by nanomedicine for effective cancer therapy
    • Authors: Rana Jahanban-Esfahlan; Miguel de la Guardia, Delshad Ahmadi, Bahman Yousefi
      First page: 2019
      Abstract: Hypoxia, a characteristic feature of tumors, is indispensable to tumor angiogenesis, metastasis, and multi drug resistance. Hypoxic avascular regions, deeply embedded inside the tumors significantly hinder delivery of therapeutic agents. The low oxygen tension results in resistance to the current applied anti‐cancer therapeutics including radiotherapy, chemotherapy, and photodynamic therapy, the efficacy of which is firmly tied to the level of tumor oxygen supply. However, emerging data indicate that nanocarriers/nanodrugs can offer substantial benefits to improve the efficacy of current therapeutics, through modulation of tumor hypoxia. This review aims to introduce the most recent advances made in nanocarrier mediated targeting of tumor hypoxia. The first part is dedicated to the approaches by which nanocarriers could be designed to target/leverage hypoxia. These approaches include i) inhibiting Hypoxia Inducer Factor (HIF‐1α); ii) hypoxia activated prodrugs/linkers; and iii) obligate anaerobe mediated targeting of tumor hypoxia. The second part, details novel nanosystems proposed to modulate tumor hypoxia through tumor oxygenation. These methods seek to lessen tumor hypoxia through vascular normalization, or reoxygenation therapy. The reoxygenation of tumor could be accomplished by: i) generation of oxygen filled nanocarriers; ii) natural/artificial oxygen nanocarriers; and iii) oxygen generators. The efficacy of each approach and their potential in cancer therapy is further discussed.Hypoxic avascular regions, deeply embedded inside the tumors significantly hinder delivery of therapeutic agents. Hypoxia could be accomplished by developing HIF‐1a and VEGF inhibitors that are now successfully integrated into the oncology practice. Incorporation of nanomedicine in targeting tumor hypoxia could dramatically enhance potential of anti‐cancer therapeutics.
      PubDate: 2017-05-03T07:05:33.005193-05:
      DOI: 10.1002/jcp.25859
  • Adenosine and adenosine receptors in the immunopathogenesis and treatment
           of cancer
    • Authors: Mohammad H. Kazemi; Sahar Raoofi Mohseni, Mohammad Hojjat-Farsangi, Enayat Anvari, Ghasem Ghalamfarsa, Hamed Mohammadi, Farhad Jadidi-Niaragh
      First page: 2032
      Abstract: Tumor cells overcome anti‐tumor responses in part through immunosuppressive mechanisms. There are several immune modulatory mechanisms. Among them, adenosine is an important factor which is generated by both cancer and immune cells in tumor microenvironment to suppress anti‐tumor responses. Two cell surface expressed molecules including CD73 and CD39 catalyze the generation of adenosine from adenosine triphosphate (ATP). The generation of adenosine can be enhanced under metabolic stress like tumor hypoxic conditions. Adenosine exerts its immune regulatory functions through four different adenosine receptors (ARs) including A1, A2A, A2B, and A3 which are expressed on various immune cells. Several studies have indicated the overexpression of adenosine generating enzymes and ARs in various cancers which was correlated with tumor progression. Since the signaling of ARs enhances tumor progression, their manipulation can be promising therapeutic approach in cancer therapy. Accordingly, several agonists and antagonists against ARs have been designed for cancer therapy. In this review, we will try to clarify the role of different ARs in the immunopathogenesis, as well as their role in the treatment of cancer.Adenosine is one of the important immunosuppressive factors in the tumor microenvironment. Therefore, modulation of adenosine receptors can be considered as the potent cancer therapeutic method.
      PubDate: 2017-05-03T07:35:31.206649-05:
      DOI: 10.1002/jcp.25873
  • Targeting RAS signaling pathway as a potential therapeutic target in the
           treatment of colorectal cancer
    • Authors: Afsane Bahrami; Seyed Mahdi Hassanian, Soodabeh ShahidSales, Zahra Farjami, Malihe Hasanzadeh, Kazem Anvari, Amir Aledavood, Mina Maftouh, Gordon A. Ferns, Majid Khazaei, Amir Avan
      First page: 2058
      Abstract: The V‐Ki‐ras2 Kirsten rat sarcoma viral oncogene homolog (KRAS) is frequently dysregulated in colorectal cancer (CRC). It is involved in the modulation of several downstream effectors, that include: Raf/Mek/Erk, PI3K/Akt, RalGDS/p38MAPK, and Rac/Rho, and thereby influences tumorigenesis, the invasive behaviors of tumor cell, and resistance to therapy. There is growing evidence exploring the use of drugs that target these pathways in the treatment of CRC. Cetuximab has been approved for CRC patients without a KRAS mutation, or for EGFR‐expressing metastatic CRC, although some of the patients have a mutation of KRAS and NRAS. This review summarizes the recent knowledge about the therapeutic potential of targeting RAS with particular emphasis on recent preclinical and clinical studies in treatment of CRC.
      PubDate: 2017-05-23T11:30:23.207418-05:
      DOI: 10.1002/jcp.25890
  • Blocking fibrotic signaling in fibroblasts from patients with carpal
           tunnel syndrome
    • Authors: Yoshiaki Yamanaka; Anne Gingery, Gosuke Oki, Tai-Hua Yang, Chunfeng Zhao, Peter C. Amadio
      First page: 2067
      Abstract: Fibrosis of the subsynovial connective tissue (SSCT) in carpal tunnel syndrome (CTS) patients is increasingly recognized as an important aspect of CTS pathophysiology. In this study, we evaluated the effect of blocking profibrotic pathways in fibroblasts from the SSCT in CTS patients. Fibroblasts were stimulated with transforming growth factor β1 (TGF‐β1), and then treated either with a specific fibrosis pathway inhibitor targeting TGF‐β receptor type 1 (TβRI), platelet‐derived growth factor receptor (PDGFR), epidermal growth factor receptor (EGFR), or vascular endothelial growth factor receptor (VEGFR). Fibrosis array and quantitative real‐time polymerase chain reaction of fibrotic genes were evaluated. Array gene expression analysis revealed significant down‐regulation of multiple fibrotic genes after treatment with TβRI, PDGFR, and VEGFR inhibitors. No array fibrotic genes were significantly down‐regulated with EGFR inhibition. Further gene expression analysis of known CTS fibrosis markers collagen type I A2 (Col1), collagen type III A1 (Col3), connective tissue growth factor (CTGF), and SERPINE1 showed significantly down‐regulation after TβRI inhibition. In contrast, VEGFR inhibition significantly down‐regulated CTGF and SERPINE1, whereas, PDGFR and EGFR inhibition significantly down‐regulated Col3. Taken together the inhibition of TβRI appears to be the primary mediator of fibrotic gene expression in fibroblasts from CTS patients. TGF‐β/Smad activity was further evaluated, and as expected inhibition of Smad activity was significantly down‐regulated after inhibition of TβRI, but not with PDGFR, VEGFR, or EGFR inhibition. These results indicate that local therapies specifically targeting TGF‐β signaling alone or in combination offer the potential of a novel local antifibrosis therapy for patients with CTS.In this study, we evaluated the effect of blocking profibrotic pathways by receptor blocking agents in fibroblasts from patients with carpal tunnel syndrome (CTS). There was significant down‐regulation of multiple fibrotic genes after treatment with a TFFβ1R (SD208) inhibitor, with smaller effects from PDGFR (AG1296) and VEGFR (Axitinib) inhibitors. These results indicate that local therapies specifically targeting TGF‐β signaling, alone or in combination with other pathways, offer the potential of a novel local anti‐fibrosis therapy for patients with CTS.
      PubDate: 2017-05-03T09:25:34.519933-05:
      DOI: 10.1002/jcp.25901
  • Pannexin hemichannels: A novel promising therapy target for oxidative
           stress related diseases
    • Authors: Jin Xu; Linxi Chen, Lanfang Li
      First page: 2075
      Abstract: Pannexins, which contain three subtypes: pannexin‐1, ‐2, and ‐3, are vertebrate glycoproteins that form non‐junctional plasma membrane intracellular hemichannels via oligomerization. Oxidative stress refers to an imbalance of the generation and elimination of reactive oxygen species (ROS). Studies have shown that elevated ROS levels are pivotal in the development of a variety of diseases. Recent studies indicate that the occurrence of these oxidative stress related diseases is associated with pannexin hemichannels. It is also reported that pannexins regulate the production of ROS which in turn may increase the opening of pannexin hemichannels. In this paper, we review recent researches about the important role of pannexin hemichannels in oxidative stress related diseases. Thus, pannexin hemichannels, novel therapeutic targets, hold promise in managing oxidative stress related diseases such as the tumor, inflammatory bowel diseases (IBD), pulmonary fibrosis, chronic obstructive pulmonary disease (COPD), cardiovascular disease, insulin resistance (IR), and neural degeneration diseases.In this review, we find that there is intimate relationship between pannexin hemichannels and oxidative stress, thus we discuss possible mechanism of pannexin hemichannels on oxidative stress related diseases.
      PubDate: 2017-05-03T09:10:39.532578-05:
      DOI: 10.1002/jcp.25906
  • Probiotics are a good choice in remission of inflammatory bowel diseases:
           A meta analysis and systematic review
    • Authors: Mahboube Ganji-Arjenaki; Mahmoud Rafieian-Kopaei
      First page: 2091
      Abstract: Altered gut bacteria and bacterial metabolic pathways are two important factors in initiation and progression of inflammatory bowel disease (IBD). However, efficacy of probiotics in remission of patients with IBD has not been characterized. This study was performed on the studies that specifically assessed the efficacy of probiotics in attaining clinical response on patients with various types of IBD. The efficacy of variant species of probiotics in different conditions and the influence of study quality in outcomes of randomized controlled trials (RCTs) were also assessed. The RCTs were collected by searching in MEDLINE Web of Science and Google scholar. Then all studies were abstracted in abstraction form and the outcomes were analyzed with fixed‐effect and mixed‐effect models for assessment of efficacy of variant species of probiotics in subgroups of IBDs. Analysis of 9 trials showed that probiotics had not significant effect on Crohn's disease (CD) (p = 0.07) but analysis of 3 trials in children with IBD revealed a significant advantage (p 
      PubDate: 2017-05-03T09:20:25.235552-05:
      DOI: 10.1002/jcp.25911
  • The diversified function and potential therapy of ectopic olfactory
           receptors in non‐olfactory tissues
    • Authors: Zhe Chen; Hong Zhao, Nian Fu, Linxi Chen
      First page: 2104
      Abstract: Olfactory receptors (ORs) are mainly distributed in olfactory neurons and play a key role in detecting volatile odorants, eventually resulting in the production of smell perception. Recently, it is also reported that ORs are expressed in non‐olfactory tissues including heart, lung, sperm, skin, and cancerous tissues. Interestingly, ectopic ORs are associated with the development of diseases in non‐olfactory tissues. For instance, ectopic ORs initiate the hypoxic ventilatory responses and maintain the oxygen homeostasis of breathing in the carotid body when oxygen levels decline. Ectopic ORs induce glucose homeostasis in diabetes. Ectopic ORs regulate systemic blood pressure by increasing renin secretion and vasodilation. Ectopic ORs participate in the process of tumor cell proliferation, apoptosis, metastasis, and invasiveness. Ectopic ORs accelerate the occurrence of obesity, angiogenesis and wound‐healing processes. Ectopic ORs affect fetal hemoglobin levels in sickle cell anemia and thalassemia. Finally, we also elaborate some ligands targeting for ORs. Obviously, the diversified function and related signal pathway of ectopic ORs may play a potential therapeutic target in non‐olfactory tissues. Thus, this review focuses on the latest research results about the diversified function and therapeutic potential of ectopic ORs in non‐olfactory tissues.This review focuses on the latest research results about the diversified function and therapeutic potential of ectopic ORs in non‐olfactory tissues.
      PubDate: 2017-05-05T09:00:24.06571-05:0
      DOI: 10.1002/jcp.25929
  • NLRP3 inflammasome: Its regulation and involvement in atherosclerosis
    • Authors: Zahra Hoseini; Fatemeh Sepahvand, Bahman Rashidi, Amirhossein Sahebkar, Aria Masoudifar, Hamed Mirzaei
      First page: 2116
      Abstract: Inflammasomes are intracellular complexes involved in the innate immunity that convert proIL‐1β and proIL‐18 to mature forms and initiate pyroptosis via cleaving procaspase‐1. The most well‐known inflammasome is NLRP3. Several studies have indicated a decisive and important role of NLRP3 inflammasome, IL‐1β, IL‐18, and pyroptosis in atherosclerosis. Modern hypotheses introduce atherosclerosis as an inflammatory/lipid‐based disease and NLRP3 inflammasome has been considered as a link between lipid metabolism and inflammation because crystalline cholesterol and oxidized low‐density lipoprotein (oxLDL) (two abundant components in atherosclerotic plaques) activate NLRP3 inflammasome. In addition, oxidative stress, mitochondrial dysfunction, endoplasmic reticulum (ER) stress, and lysosome rupture, which are implicated in inflammasome activation, have been discussed as important events in atherosclerosis. In spite of these clues, some studies have reported that NLRP3 inflammasome has no significant effect in atherogenesis. Our review reveals that some molecules such as JNK‐1 and ASK‐1 (upstream regulators of inflammasome activation) can reduce atherosclerosis through inducing apoptosis in macrophages. Notably, NLRP3 inflammasome can also cause apoptosis in macrophages, suggesting that NLRP3 inflammasome may mediate JNK‐induced apoptosis, and the apoptotic function of NLRP3 inflammasome may be a reason for the conflicting results reported. The present review shows that the role of NLRP3 in atherogenesis can be significant. Here, the molecular pathways of NLRP3 inflammasome activation and the implications of this activation in atherosclerosis are explained.
      PubDate: 2017-05-23T11:30:29.031805-05:
      DOI: 10.1002/jcp.25930
  • Roles of neurotrophins in skeletal tissue formation and healing
    • Authors: Yu-Wen Su; Xin-Fu Zhou, Bruce K. Foster, Brian L. Grills, Jiake Xu, Cory J. Xian
      First page: 2133
      Abstract: Neurotrophins and their receptors are key molecules that are known to be critical in regulating nervous system development and maintenance and have been recognized to be also involved in regulating tissue formation and healing in skeletal tissues. Studies have shown that neurotrophins and their receptors are widely expressed in skeletal tissues, implicated in chondrogenesis, osteoblastogenesis, and osteoclastogenesis, and are also involved in regulating tissue formation and healing events in skeletal tissue. Increased mRNA expression for neurotrophins NGF, BDNF, NT‐3, and NT‐4, and their Trk receptors has been observed in injured bone tissues, and NT‐3 and its receptor, TrkC, have been identified to have the highest induction at the injury site in a drill‐hole injury repair model in both bone and the growth plate. In addition, NT‐3 has also recently been shown to be both an osteogenic and angiogenic factor, and this neurotrophin can also enhance expression of the key osteogenic factor, BMP‐2, as well as the major angiogenic factor, VEGF, to promote bone formation, vascularization, and healing of the injury site. Further studies, however, are needed to investigate if different neurotrophins have differential roles in skeletal repair, and if NT‐3 can be a potential target of intervention for promoting bone fracture healing.Increasing evidence now suggests that neurotrophins also have important roles in bone. Among the neurotrophins and receptors, the NT‐3/TrkC signaling (induced far more prominently compared to other NTs and other Trk receptors) may potentially play a prominent role in the bone healing.
      PubDate: 2017-05-23T12:05:28.79698-05:0
      DOI: 10.1002/jcp.25936
  • Cytoskeletal remodeling and regulation of cell fate in the hypertensive
           neonatal pulmonary artery in response to stress
    • Authors: Dina Johar
      First page: 2146
      Abstract: Neonatal pulmonary hypertension (PHN) is a lethal progressive disease that occurs in prenatal circulatory transition. Mechanical wall strain caused by cardiac pulsation integrates with hypoxia to generate rapidly progressive myocyte cytoskeleton disassembly and failure to exert force generation. The physiological responses to such an interaction have not been investigated. The persistent phenotype does not respond to traditional vasodilator therapy; hence, there is a need for new treatment strategies to improve the morbidity and mortality outcomes. We reviewed the current research methods, models, and markers of persistent PHN relevant to oxidative and nitrosative stress as well as cell fate commitment, with an emphasis on apoptosis and proliferation. We surveyed potential investigations into the role of senescence in neonatal PHN cell fate decision programming during vasodilator treatment and suggested putative drug targets to improve clinical outcomes. We identified important signaling intermediates of senescence and cell cycle entry regulation in hypertensive pulmonary arterial tissues.Identifying the concerted interplay between ROS/RNS generation, senescence, and inflammatory signaling will lead to therapeutic targets to combat vascular remodeling in PHN; the factor most limiting to treatment responses is irreversible fibrotic thickening. Prevention of the latter will help reduce mortality and morbidity in infants with PHN and prolong the window of opportunity for vasodilator therapies.
      PubDate: 2017-06-15T04:10:27.053859-05:
      DOI: 10.1002/jcp.25950
  • The therapeutic potential of targeting the BRAF mutation in patients with
           colorectal cancer
    • Authors: Afsane Bahrami; AmirReza Hesari, Majid Khazaei, Seyed Mahdi Hassanian, Gordon A. Ferns, Amir Avan
      First page: 2162
      Abstract: Colorectal cancer is among the most lethal malignancies globally. BRAF is a member of the RAS/RAF/MEK/ERK signaling pathway. Its constitutive activation can result in increased cellular growth, development, invasion, and resistance to therapy. A mutation of the BRAF gene is present in 5–10% of metastatic colorectal cancers. BRAF mutations have been found to predict a lack of benefit to anti‐EGFR therapy in metastatic CRC. Furthermore, CRC containing the BRAF V600E mutation display an innate resistance to BRAF inhibitors. The mechanisms of cell resistance can be explained at least in part by ERK dependent and ERK in‐dependent pathway. Clinical trials evaluating the combinations of BRAF, PI3K, EGFR, and/or MEK inhibitors have revealed promising activity in BRAF mutant containing CRCs. There may be some benefit from future studies that focus on improving the efficacy of combined therapy in CRC with respect to the sustained effects. The aim of current review is to give an overview about the current status and prospective regarding the therapeutic potential of targeting BRAF mutant colorectal cancer.
      PubDate: 2017-05-23T11:30:25.231009-05:
      DOI: 10.1002/jcp.25952
  • Anti‐tumor effects of crocetin and related molecular targets
    • Authors: Maliheh Moradzadeh; Hamid Reza Sadeghnia, Alijan Tabarraei, Amirhossein Sahebkar
      First page: 2170
      Abstract: Natural products have gained a wide popularity as chemopreventive and anti‐cancer agents owing to their multi‐mechanistic mode of action, availability and synergism with several conventional chemotherapeutic agents. Crocetin is a carotenoid compound isolated from the stigma of Crocus sativus L. (saffron). Crocetin has shown promising effects as an anti‐tumor agent in animal models and cell culture systems. Crocetin retards the growth of cancer cells via inhibiting nucleic acid synthesis, enhancing anti‐oxidative system, and inducing apoptosis and differentiation pathways. The present review outlines natural sources of crocetin, and its pharmacokinetic and pharmacological properties relevant to the prevention and treatment of cancer. Also, we discuss molecular targets underlying the putative anti‐tumor effects of crocetin.The present review outlines natural sources of crocetin, and its pharmacokinetic and pharmacological properties relevant to the prevention and treatment of cancer. Also, we discuss molecular targets underlying the putative anti‐tumor effects of crocetin.
      PubDate: 2017-06-07T05:53:00.589784-05:
      DOI: 10.1002/jcp.25953
  • Therapeutic potential of novel formulated forms of curcumin in the
           treatment of breast cancer by the targeting of cellular and physiological
           dysregulated pathways
    • Authors: Amir Tajbakhsh; Malihe Hasanzadeh, Mehdi Rezaee, Mostafa Khedri, Majid Khazaei, Soodabeh ShahidSales, Gordon A. Ferns, Seyed Mahdi Hassanian, Amir Avan
      First page: 2183
      Abstract: Breast cancer is among the most important causes of cancer related death in women. There is a need for novel agents for targeting key signaling pathways to either improve the efficacy of the current therapy, or reduce toxicity. There is some evidence that curcumin may have antitumor activity in breast cancer. Several clinical trials have investigated its activity in patients with breast cancer, including a recent trial in breast cancer patients receiving radiotherapy, in whom it was shown that curcumin reduced the severity of radiation dermatitis, although it is associated with low bioavailability. Several approaches have been developed to increase its absorption rate (e.g., nano crystals, liposomes, polymers, and micelles) and co‐delivery of curcumin with adjuvants as well as different conjugation to enhance its bioavailability. In particular, micro‐emulsions is an option for transdermal curcumin delivery, which has been reported to increase its absorption. Lipid‐based nano‐micelles is another approach to enhance curcumin absorption via gastrointestinal tract, while polymer‐based nano‐formulations (e.g., poly D, L‐lactic‐co‐glycolic [PLGA]) allows the release of curcumin at a sustained level. This review summarizes the current data of the therapeutic potential of novel formulations of curcumin with particular emphasis on recent preclinical and clinical studies in the treatment of breast cancer.
      PubDate: 2017-05-23T11:25:23.585815-05:
      DOI: 10.1002/jcp.25961
  • Genetic variants as potential predictive biomarkers in advanced colorectal
           cancer patients treated with oxaliplatin‐based chemotherapy
    • Authors: Afsane Bahrami; Forouzan Amerizadeh, Seyed Mahdi Hassanian, Soodabeh ShahidSales, Majid Khazaei, Mina Maftouh, Majid Ghayour-Mobarhan, Gordon A. Ferns, Amir Avan
      First page: 2193
      Abstract: Chemotherapy regimen containing oxaliplatin is often the first‐line treatment for patient with advanced colorectal cancer. Oxaliplatin binds to DNA, leading to the formation of crosslinks and bulky adducts. Approximately 50% of patients with CRC benefit from treatment with oxaliplatin. It is possible that genetic variants in biological pathways involved in drug transportation, drug metabolism, DNA damage repair, and cell cycle modulation might affect the activity, or efficacy of oxaliplatin. Because oxaliplatin resistance may be related to these genetic variants and may therefore be an important reason for treatment failure, we have summarized the genetic variations that have been reported to be predictive markers of the response to oxaliplatin based therapy in patients with advanced CRC.
      PubDate: 2017-05-23T11:25:27.7492-05:00
      DOI: 10.1002/jcp.25966
  • Endogenous bioelectric currents promote differentiation of the mammalian
    • Authors: Lin Cao; Jie Liu, Jin Pu, J. Martin Collinson, John V. Forrester, Colin D. McCaig
      First page: 2202
      Abstract: The functional roles of bioelectrical signals (ES) created by the flow of specific ions at the mammalian lens equator are poorly understood. We detected that mature, denucleated lens fibers expressed high levels of the α1 and β1 subunits of Na+/K+‐ATPase (ATP1A1 and ATP1B1 of the sodium pump) and had a hyperpolarized membrane potential difference (Vmem). In contrast, differentiating, nucleated lens fiber cells had little ATP1A1 and ATP1B1 and a depolarized Vmem. Mimicking the natural equatorial ES with an applied electrical field (EF) induced a striking reorientation of lens epithelial cells to lie perpendicular to the direction of the EF. An EF also promoted the expression of β‐crystallin, aquaporin‐0 (AQP0) and the Beaded Filament Structural Protein 2 (BFSP2) in lens epithelial cells (LECs), all of which are hallmarks of differentiation. In addition, applied EF activated the AKT and CDC2 and inhibition of AKT reduced the activation of CDC2. Our results indicate that the endogenous bioelectrical signal at the lens equator promotes differentiation of LECs into denucleated lens fiber cells via depolarization of Vmem. Development of methods and devices of EF application or amplification in vivo may supply a novel treatment for lens diseases and even promote regeneration of a complete new lens following cataract surgery.Mature, denucleated lens fibers expressed high levels of the α1 and β1 subunits of Na+/K+‐ATPase In contrast, differentiating, nucleated lens fiber cells had little ATP1A1 and ATP1B1. An applied electrical field (EF) induced a striking reorientation of lens epithelial cells to lie perpendicular to the direction of the EF. The applied EF activated the AKT and CDC2 and inhibition of AKT reduced the activation of CDC2.
      PubDate: 2017-08-30T05:40:55.843723-05:
      DOI: 10.1002/jcp.26074
  • Yes-associated protein 1 promotes the differentiation and mineralization
           of cementoblast
    • Authors: Beining Yang; Hualing Sun, Fangfang Song, Yanru Wu, Jiawei Wang
      First page: 2213
      Abstract: Yes-associated protein 1 (YAP1) transcriptional coactivator is a mediator of mechanosensitive signaling. Cementum, which covers the tooth root surface, continuously senses external mechanical stimulation. Cementoblasts are responsible for the mineralization and maturation of the cementum. However, the effect of YAP1 on cementoblast differentiation remains largely unknown. In this study, we initially demonstrated that YAP1 overexpression enhanced the mineralization ability of cementoblasts. YAP1 upregulated the mRNA and protein expression of several cementogenesis markers, such as alkaline phosphatase (ALP), runt-related transcription factor 2 (Runx2), osteocalcin (OCN), and dentin matrix acidic phosphoprotein 1 (DMP1). The YAP1 overexpression group showed higher intensities of ALP and Alizarin red stain than the YAP1-knockdown group. Unexpectedly, a sharp increase in the expression of dentin sialophosphoprotein (DSPP) was induced by the overexpression of YAP1. Knockdown of YAP1 suppressed DSPP transcriptional activity. YAP1 overexpression activated Smad-dependent BMP signaling and slightly inhibited Erk1/2 signaling pathway activity. Treatment with specific BMP antagonist (LDN193189) prevented the upregulation of the mRNA levels of ALP, RUNX2, and OCN, as well as intensity of ALP-stained and mineralized nodules in cementoblasts. The Erk1/2 signaling pathway inhibitor (PD 98,059) upregulated these cementogenesis markers. Thus, our study suggested that YAP1 enhanced cementoblast mineralization in vitro. YAP1 exerted its effect on the cementoblast partly by regulating the Smad-dependent BMP and Erk1/2 signaling pathways. This article is protected by copyright. All rights reserved
      PubDate: 2017-09-25T04:51:02.534387-05:
      DOI: 10.1002/jcp.26089
  • Triptolide prevents extracellular matrix accumulation in experimental
           diabetic kidney disease by targeting microRNA‐137/Notch1 pathway
    • Authors: Fei Han; Shanshan Wang, Yunpeng Chang, Chunjun Li, Juhong Yang, Zhe Han, Baocheng Chang, Bei Sun, Liming Chen
      First page: 2225
      Abstract: MicroRNAs (miRNAs) are involved in multiple biological functions via suppressing target genes. Triptolide is a monomeric compound isolated from a traditional Chinese herb, which exerts protective roles in many kinds of glomerular diseases. However, our understanding of the triptolide effect on miRNAome is still limited. In this study, we found that triptolide significantly decreased albuminuria and improved glomerulosclerosis in rats with diabetic kidney disease (DKD). And triptolide also inhibited extracellular matrix (ECM) protein accumulation and the notch1 pathway activation under diabetic conditions. MiR‐137 was significantly decreased in the HG (high glucose)‐treated HRMCs and in the kidney tissues of the diabetic rats, but was upregulated by triptolide. In addition, overexpression of miR‐137 exerted similar effects to those of triptolide, while miR‐137 inhibition aggravated ECM protein accumulation. Luciferase reporter assay results demonstrated that miR‐137 directly targets Notch1. Furthermore, the miR‐137‐dependent effects were due to Notch1 suppression that in turn inhibited ECM protein expression, key mediators of glomerulosclerosis. Finally, downregulation of miR‐137 reversed the ECM inhibition role of triptolide in HG cultured HRMCs. Taken together, these findings indicate that triptolide is a potential therapeutic option for DKD and that miR‐137/Notch1 pathway play roles in the anti‐glomerulosclerosis mechanism of triptolide.Diabetic condition decreases the miR‐137 levels in renal mesangial cells. MiR‐137 decreases the expression of Col IV and FN by targeting Notch1. On the other hand, triptolide may protect DKD against glomerulosclerosis by targeting miR‐137/Notch1 pathway.
      PubDate: 2017-09-13T05:46:19.910411-05:
      DOI: 10.1002/jcp.26092
  • Maresin 1 inhibits TNF‐alpha‐induced lipolysis and autophagy
           in 3T3‐L1 adipocytes
    • Authors: Laura M Laiglesia; Silvia Lorente-Cebrián, Miguel López-Yoldi, Raquel Lanas, Neira Sáinz, Jose Alfredo Martínez, Maria J. Moreno-Aliaga
      First page: 2238
      Abstract: Obesity is associated with high levels of pro‐inflammatory cytokines such as tumor necrosis factor‐alpha (TNF‐α), which promotes inflammation in adipose tissue. The omega‐3 PUFAs, and their derived lipid mediators, such as Maresin 1 (MaR1) have anti‐inflammatory effects on adipose tissue. This study aimed to analyze if MaR1 may counteract alterations induced by TNF‐α on lipolysis and autophagy in mature 3T3‐L1 adipocytes. Our data revealed that MaR1 (1–100 nM) inhibited the TNF‐α‐induced glycerol release after 48 hr, which may be related to MaR1 ability of preventing the decrease in lipid droplet‐coating protein perilipin and G0/G1 Switch 2 protein expression. MaR1 also reversed the decrease in total hormone sensitive lipase (total HSL), and the ratio of phosphoHSL at Ser‐565/total HSL, while preventing the increased ratio of phosphoHSL at Ser‐660/total HSL and phosphorylation of extracellular signal‐regulated kinase 1/2 induced by TNF‐α. Moreover, MaR1 counteracted the cytokine‐induced decrease of p62 protein, a key autophagy indicator, and also prevented the induction of LC3II/LC3I, an important autophagosome formation marker. Current data suggest that MaR1 may ameliorate TNF‐α‐induced alterations on lipolysis and autophagy in adipocytes. This may also contribute to the beneficial actions of MaR1 on adipose tissue and insulin sensitivity in obesity.MaR1 counteracts the changes induced by TNF‐α on some of the main lipases and lipid droplets proteins controlling lipolysis (perilipin, HSL, and G0S2) as well as on the proteins regulating autophagy (p62 and LC3). These observations suggest that MaR1 may represent a promising therapeutic agent to counteract the alterations induced by inflammation in adipose tissue.
      PubDate: 2017-08-30T05:27:24.166208-05:
      DOI: 10.1002/jcp.26096
  • The xenoestrogens biphenol‐A and nonylphenol differentially regulate
           metalloprotease‐mediated shedding of EGFR ligands
    • Authors: Paulina Urriola-Muñoz; Xue Li, Thorsten Maretzky, David R. McIlwain, Tak W. Mak, Juan G. Reyes, Carl P. Blobel, Ricardo D. Moreno
      First page: 2247
      Abstract: The xenoestrogens bisphenol‐A (BPA) and nonylphenol (NP) are endocrine disruptors used in the plastic polymer industry to manufacture different products for human use. Previous studies have suggested a role of these compounds in the shedding of signaling molecules, such as tumor necrosis factor α (TNF‐α). The aim of this work was to evaluate the effect of BPA and NP on the sheddase ADAM17 and its newly discovered regulators iRhom1 and iRhom2 in the release of EGFR‐ligands. We report that BPA and NP can stimulate the release of the ADAM17‐substrates HB‐EGF and TGF‐α. In cells lacking ADAM17 (Adam17−/− mEFs) BPA‐stimulated release of HB‐EGF, but not TGF‐α, was strongly reduced, whereas NP‐stimulated shedding of HB‐EGF and TGF‐α was completely abolished. Inactivation of both ADAM17 and the related ADAM10 (Adam10/17−/− mEFs) completely prevented the release of these substrates. In the absence of iRhom1, BPA‐ or NP‐stimulated release of HB‐EGF or TGF‐α was comparable to wild‐type control mEFs, conversely the BPA‐induced release of HB‐EGF was abolished in iRhom2−/− mEFs. The defect in shedding of HB‐EGF in iRhom2−/− mEF cells could be rescued by overexpressing iRhom2. Interestingly, the NP‐stimulated release of HB‐EGF was not affected by the absence of iRhom2, suggesting that NP could potentially activate both ADAM10 and ADAM17. We tested this hypothesis using betacellulin (BTC), an EGFR‐ligand that is a substrate for ADAM10. We found that NP, but not BPA stimulated the release of BTC in Adam17−/−, iRhom2−/−, or iRhom1/2−/−, but not in Adam10/17−/− cells. Taken together, our results suggest that BPA and NP stimulate the release of EGFR‐ligands by differentially activating ADAM17 or ADAM10. The identification of specific effects of these endocrine disruptors on ADAM10 and ADAM17 will help to provide a better understanding of their roles in cell signaling and proinflammatory processes, and provide new potential targets for treatment of reproductive or inflammatory diseases such as asthma or breast cancer that are promoted by xenoestrogens.BPA and NP stimulate the release of EGFR‐ligands by differentially activating ADAM17 or ADAM10.
      PubDate: 2017-08-25T05:45:50.607301-05:
      DOI: 10.1002/jcp.26097
  • The depletion of MARVELD1 leads to murine placenta accreta via integrin
           β4‐dependent trophoblast cell invasion
    • Authors: Yue Chen; Hui Zhang, Fang Han, Lei Yue, Chunxiao Qiao, Yao Zhang, Peng Dou, Weizhe Liu, Yu Li
      First page: 2257
      Abstract: The placenta is a remarkable organ, it serves as the interface between the mother and the fetus. Proper invasion of trophoblast cells is required for a successful pregnancy. Previous studies have found that the adhesion molecule integrin β4 plays important roles during trophoblast cell invasion. Here, we found that the overall birth rate of the MARVELD1 knockout mouse is much lower than that of the wild‐type mouse (p 
      PubDate: 2017-08-17T06:51:39.06216-05:0
      DOI: 10.1002/jcp.26098
  • FHOD1 regulates cytoplasmic actin‐based spindle migration for mouse
           oocyte asymmetric cell division
    • Authors: Meng-Hao Pan; Fei Wang, Yujie Lu, Feng Tang, Xing Duan, Yu Zhang, Bo Xiong, Shao-Chen Sun
      First page: 2270
      Abstract: FHOD1 is a member of Diaphanous‐related formins (DRFs) which belongs to the Formin family. Previous studies have shown that the DFRs might affect several cellular functions such as morphogenesis, cytokinesis, cell polarity, and embryonic differentiation. However, there is no evidence showing the functions of FHOD1 during oocyte meiosis. This study is aimed at exploring the roles of FHOD1 during the mammalian oocyte maturation. Immunofluorescent staining showed that FHOD1 was restricted to the nucleus in germinal vesicle (GV) stage of the oocytes, after the GV breakdown FHOD1 was primarily located at two poles of the spindle at both metaphases I and II stages. Knockdown of FHOD1 by siRNA injection did not affect polar body extrusion but generated the large polar bodies. In addition, we observed the spindle migration failure in metaphase I oocytes, with a large number of meiotic spindles anchoring in the center of cytoplasm. The expression level of cytoplasmic actin but not cortex actin was significantly reduced, indicating that FHOD1 regulates cytoplasmic actin distribution for the spindle movement. Furthermore, we found that the disruption of ROCK (the Rho‐dependent protein kinase) with inhibitor Y‐27632 caused the decreased FHOD1 protein expression. Therefore, our data indicate that FHOD1 is regulated by ROCK for cytoplasm actin assembly and spindle migration during mouse oocyte meiosis.FHOD1 mainly localized at the poles of meiotic spindle in oocytes, and FHOD1 regulated cytoplasmic actin filaments for spindle migration. FHOD1 is a new regulator for oocyte asymmetric cell division.
      PubDate: 2017-08-17T06:16:05.941968-05:
      DOI: 10.1002/jcp.26099
  • Aquaporin1 and 3 modification as a result of chondrogenic differentiation
           of human mesenchymal stem cell
    • Authors: Adriana C. E. Graziano; Rosanna Avola, Giovanna Pannuzzo, Venera Cardile
      First page: 2279
      Abstract: Chondrocytes are cells of articular cartilage particularly sensitive to water transport and ionic and osmotic changes from extracellular environment and responsible for the production of the synovial fluid. Aquaporins (AQPs) are a family of water and small solute transport channel proteins identified in several tissues, involved in physiological pathways and in manifold human diseases. In a recent period, AQP1 and 3 seem to have a role in metabolic water regulation in articular cartilage of load bearing joints. The aim of this study was to examine the levels of AQP1 and 3 during the chondrogenic differentiation of human mesenchymal stem cells (MSCs) derived from adipose tissue (AT). For the determination of chondrogenic markers and AQPs levels, glycosaminoglycans (GAGs) quantification, immunocytochemistry, RT‐PCR, and Western blot were used after 0, 7, 14, 21, and 28 days from the start of differentiation. At 21 days, chondrocytes derived from AT‐MSCs were able to produce augmented content of GAGs and significant quantity of SOX‐9, lubricin, aggrecan, and collagen type II, suggesting hyaline cartilage formation, in combination with an increase of AQP3 and AQP1. However, while AQP1 level decreased after 21 days; AQP3 reached higher values at 28 days. The expression of AQP1 and 3 is a manifestation of physiological adaptation of functionally mature chondrocytes able to respond to the change of their internal environment influenced by extracellular matrix. The alteration or loss of expression of AQP1 and 3 could contribute to destruction of chondrocytes and to development of cartilage damage.We examined the levels of AQP1 and 3 during the chondrogenic differentiation of human mesenchymal stem cells (MSCs) derived from adipose tissue (AT). At 21 days, chondrocytes derived from AT‐MSCs were able to produce augmented content of GAGs and significant quantity of SOX‐9, lubricin, aggrecan and collagen type II, suggesting hyaline cartilage formation, In parallel, the same cells increased AQP3 and AQP1. AQP1 and AQP3 can be functionally involved in homeostasis and chondrogenic differentiation of AT‐MSCs.
      PubDate: 2017-08-28T05:45:46.163236-05:
      DOI: 10.1002/jcp.26100
  • miR‐199a‐3p is involved in estrogen‐mediated autophagy through the
           IGF‐1/mTOR pathway in osteocyte‐like MLO‐Y4 cells
    • Authors: Jiayao Fu; Lingyu Hao, Yawen Tian, Yang Liu, Yijing Gu, Junhua Wu
      First page: 2292
      Abstract: To date, evidence indicates that estrogen partially modulates cellular processes through microRNAs. Autophagy is a catabolic process that is regulated by multiple factors and is associated with skeletal diseases. However, whether estrogen regulates osteocyte autophagy via microRNAs is largely unknown. In this study, we observed the up‐regulation of microRNA‐199a‐3p, a post‐transcriptional regulatory factor, in osteocytic areas in ovariectomized (OVX) mice. The mature forms of miR‐199a‐3p and pri‐miR‐199a were produced in response to estrogen signaling in osteocyte‐like MLO‐Y4 cells. Western blotting, autophagic flux detection, mRFP‐GFP‐LC3 fluorescence, and electron microscopy confirmed that miR‐199a‐3p induced autophagy in MLO‐Y4 cells, although cellular apoptosis was not affected. Additionally, we documented the ability of estrogen to mediate osteocyte autophagy. Based on our in vivo data, estrogen deficiency induced autophagy in osteocytes. Treatment of starved MLO‐Y4 cells with 17β‐estradiol suppressed the excess autophagy induced by starvation via activation of mammalian target of rapamycin (mTOR)‐related signaling cascades, while administration of rapamycin reversed the effects of 17β‐estradiol. Meanwhile, miR‐199a‐3p overexpression reversed 17β‐estradiol‐mediated regulation of autophagy in MLO‐Y4 cells. According to mechanistic studies, miR‐199a‐3p inhibited the mTOR pathway by directly binding to the 3′‐untranslated regions of insulin growth factor‐1 (IGF‐1) and mTOR. However, overexpression of miR‐199a‐3p inhibited IGF‐1 phosphorylation and mTOR‐related pathways. Knockdown of mTOR and IGF‐1 abolished estrogen signaling and restored LC3‐II expression through mTOR re‐activation, respectively. Thus, miR‐199a‐3p appears to be involved in the estrogen regulatory networks that mediate bone cell autophagy, potentially by targeting IGF‐1 and mTOR.miR‐199a‐3p is mediated by estrogen signaling in osteocyte both in vivo and in vitro. miR‐199a‐3p induces autophagy in MLO‐Y4 cells via directly binding to IGF‐1 and mTOR.
      PubDate: 2017-08-25T05:55:49.132212-05:
      DOI: 10.1002/jcp.26101
  • Differential protein modulation by ketoprofen and ibuprofen underlines
           different cellular response by gastric epithelium
    • Authors: Laura Brandolini; Michele d'Angelo, Andrea Antonosante, Sara Villa, Loredana Cristiano, Vanessa Castelli, Elisabetta Benedetti, Mariano Catanesi, Andrea Aramini, Alberto Luini, Seetharaman Parashuraman, Emilia Mayo, Antonio Giordano, Annamaria Cimini, Marcello Allegretti
      First page: 2304
      Abstract: Ketoprofen L‐lysine salt (KLS), is widely used due to its analgesic efficacy and tolerability, and L‐lysine was reported to increase the solubility and the gastric tolerance of ketoprofen. In a recent report, L‐lysine salification has been shown to exert a gastroprotective effect due to its specific ability to counteract the NSAIDs‐induced oxidative stress and up‐regulate gastroprotective proteins. In order to derive further insights into the safety and efficacy profile of KLS, in this study we additionally compared the effect of lysine and arginine, another amino acid counterion commonly used for NSAIDs salification, in control and in ethanol challenged human gastric mucosa model. KLS is widely used for the control of post‐surgical pain and for the management of pain and fever in inflammatory conditions in children and adults. It is generally well tolerated in pediatric patients, and data from three studies in >900 children indicate that oral administration is well tolerated when administered for up to 3 weeks after surgery. Since only few studies have so far investigated the effect of ketoprofen on gastric mucosa maintenance and adaptive mechanisms, in the second part of the study we applied the cMap approach to compare ketoprofen‐induced and ibuprofen‐induced gene expression profiles in order to explore compound‐specific targeted biological pathways. Among the several genes exclusively modulated by ketoprofen, our attention was particularly focused on genes involved in the maintenance of gastric mucosa barrier integrity (cell junctions, morphology, and viability). The hypothesis was further validated by Real‐time PCR.Ketoprofen and ibuprofen differentially modulate proteins involved in gastroprotection.
      PubDate: 2017-08-25T05:45:58.443799-05:
      DOI: 10.1002/jcp.26102
  • Neoadjuvant Chemotherapy In Triple-Negative Breast Cancer: A Multicentric
           Retrospective Observational Study In Real-Life Setting
    • Authors: Teresa Gamucci; Laura Pizzuti, Isabella Sperduti, Lucia Mentuccia, Angela Vaccaro, Luca Moscetti, Paolo Marchetti, Luisa Carbognin, Andrea Michelotti, Laura Iezzi, Alessandra Cassano, Antonino Grassadonia, Antonio Astone, Andrea Botticelli, Emanuela Magnolfi, Luigi Di Lauro, Domenico Sergi, Paola Fuso, Nicola Tinari, Maddalena Barba, Marcello Maugeri-Saccà, Elisabetta Landucci, Francesca Conti, Giuseppe Sanguineti, Michele De Tursi, Gianni Iafrate, Antonio Giordano, Gennaro Ciliberto, Clara Natoli, Patrizia Vici
      First page: 2313
      Abstract: We aimed to assess the efficacy of neoadjuvant chemotherapy (NACT) in a cohort of 213 triple-negative breast cancer (TNBC) patients treated in real-world practice at 8 Italian cancer centres. We computed descriptive statistics for all the variable of interest. Factors testing significant in univariate analysis were included in multivariate models. Survival data were compared by Kaplan-Meier curves and log-rank test.The median follow-up was 45 months. We observed 60 (28.2%) pathological complete response (pCR). The sequential anthracyclines-taxanes-based regimens produced the highest rate of pCR (42.6%), followed by concomitant anthracycline-taxane (24.2%), and other regimens (15.6%) (p = 0.008). When analyzing the role of baseline Ki-67, a 50% cut-off was the optimal threshold value for pCR prediction (p = 0.0005). The 5-year disease-free survival (DFS) was 57.3% and the 5-year overall survival (OS) was 70.8%. In patients not achieving pCR, the optimal Ki-67 variation between biopsy and surgical specimen with prognostic relevance on long-term outcomes was 13% (p = 0.04). Patients with a Ki-67 reduction (rKi-67) 6 cycles compared with their counterparts, p = 0.02). In multivariate analysis, node status, grading and bio-pathological treatment response (including pCR and rKi-67) impacted DFS and OS. Our results confirmed the advantage conferred by more than 6 cycles of a sequential antracycline-taxane-based NACT. Higher baseline Ki-67 values shows greater predictive significance on pathogical response, while the rKi-67 plays a prognostic role on long-term outcomes. This article is protected by copyright. All rights reserved
      PubDate: 2017-09-27T03:35:59.817375-05:
      DOI: 10.1002/jcp.26103
  • Primary familial brain calcification with a novel SLC20A2 mutation:
           Analysis of PiT‐2 expression and localization
    • Authors: Ilaria Taglia; Patrizia Formichi, Carla Battisti, Giulia Peppoloni, Melissa Barghigiani, Alessandra Tessa, Antonio Federico
      First page: 2324
      Abstract: Primary familial brain calcification (PFBC) is an autosomal dominant rare disorder characterized by bilateral and symmetric brain calcifications, and neuropsychiatric manifestations. Four genes have been linked to PFBC: SLC20A2, PDGFRB, PDGFB, and XPR1. In this study, we report molecular and clinical data of a PFBC patient carrying a novel SLC20A2 mutation and we investigate the impact of the mutation on PiT‐2 expression and function. Sanger sequencing of SLC20A2, PDGFRB, PDGFB, XPR1 led to the identification of a novel duplication of twelve nucleotides (c.1876_1887dup/ p.Trp626_Thr629dup) in SLC20A2 gene. SLC20A2 encodes for a cell membrane transporter (PiT‐2) involved in maintenance of inorganic phosphate homeostasis. We performed an analysis of expression and functionality of PiT‐2 protein in patient primary cultured fibroblasts. In patient fibroblasts, the mutation does not affect PiT‐2 expression but alter sub‐cellular localization. The Pi‐uptake assay revealed a less Pi depletion in patient than in control fibroblasts, suggesting that SLC20A2 duplication may impair Pi internalization. This is the first study reporting sub‐cellular expression analysis of mutant PiT‐2 in primary cultured fibroblasts from a PFBC patient, showing that p.Trp626_Thr629dup in SLC20A2 alters PiT‐2 sub‐cellular localization and reduces Pi‐uptake, leading to onset of PFBC in our patient.Primary familial brain calcification (PFBC) is an autosomal dominant rare disorder characterized by brain calcifications and neuropsychiatric manifestations. In this study, we report molecular and clinical data of a PFBC patient carrying a novel duplication of twelve nucleotides (c.1876_1887dup/ p.Trp626_Thr629dup) in SLC20A2 gene. SLC20A2 encodes for a cell membrane transporter (PiT‐2) involved in maintenance of inorganic phosphate homeostasis. The mutation does not affect PiT‐2 expression but alter sub‐cellular localization and impair Pi internalization.
      PubDate: 2017-09-18T04:20:44.841782-05:
      DOI: 10.1002/jcp.26104
  • Cyanidin suppresses autophagic activity regulating chondrocyte
           hypertrophic differentiation
    • Authors: Zhen Cao; Song Huang, Ce Dou, Qiang Xiang, Shiwu Dong
      First page: 2332
      Abstract: Cartilage is a kind of special connective tissue which does not contain neither blood vessels nor lymphatics and nerves. Therefore, the damage in cartilage is difficult to be repaired spontaneously. Constructing tissue engineered cartilage provides a new technique for cartilage repairing. Mesenchymal stem cells (MSCs) possess a unique capability of self‐renew and can differentiate into pre‐chondrocytes which are frequently applied as seed cells in tissue engineering. However, in regenerated cartilage the chondrocytes derived from MSCs can hardly maintain homeostasis and preferentially present hypertrophic like phenotype. We investigated the effects of cyanidin, a natural organic compound, on chondrogenic and subsequent hypertrophic differentiation of MSCs in order to seek approaches to inhibit chondrocyte hypertrophy. We evaluated the effects of cyanidin on expression of chondrogenic and hypertrophic marker genes through RT‐PCR, Western blot, alcian blue staining, and immunocytochemistry. The results showed that both chondrogenic related genes Sox9, Col2a1, and hypertrophic marker genes Runx2, Col10a1 were inhibited by cyanidin. In addition, we found that cyanidin promoted Nrf2 and p62 expression and suppressed LC3B expression during chondrogenic stage of MSCs. Meanwhile phosphorylation of IκBα and autophagosome related protein LC3B were inactivated by cyanidin during chondrocyte hypertrophic stage. Furthermore, rapamycin, an autophagy activator, abrogated the inhibitory effect of cyanidin on chondrogenic, and hypertrophic differentiation of MSCs. In conclusion, one potential mechanism of cyanidin, by which the chondrogenic and hypertrophic differentiation of MSCs were inhibited, was due to decreased autophagy activity. Our results indicated that cyanidin was a potential therapeutic agent for keeping mature chondrocyte functions.Cyanidin increased Nrf2 expression and inactivated autophagy to inhibit chondrogenic differentiation of MSCs. Besides, cyanidin inhibited NF‐κB signaling and suppressed autophagy to delay chondrocyte hypertrophic differentiation.
      PubDate: 2017-08-25T05:55:58.818922-05:
      DOI: 10.1002/jcp.26105
  • Structure‐based release analysis of the JC virus agnoprotein regions: A
           role for the hydrophilic surface of the major alpha helix domain in
    • Authors: A. Sami Saribas; Martyn K. White, Mahmut Safak
      First page: 2343
      Abstract: Agnoprotein (Agno) is an important regulatory protein of JC virus (JCV), BK virus (BKV) and simian virus 40 (SV40) and these viruses are unable to replicate efficiently in the absence of this protein. Recent 3D‐NMR structural data revealed that Agno contains two alpha‐helices (a minor and a major) while the rest of the protein adopts an unstructured conformation (Coric et al., 2017, J Cell Biochem). Previously, release of the JCV Agno from the Agno‐positive cells was reported. Here, we have further mapped the regions of Agno responsible for its release by a structure‐based systematic mutagenesis approach. Results revealed that amino acid residues (Lys22, Lys23, Phe31, Glu34, and Asp38) located either on or adjacent to the hydrophilic surface of the major alpha‐helix domain of Agno play critical roles in release. Additionally, Agno was shown to strongly interact with unidentified components of the cell surface when cells are treated with Agno, suggesting additional novel roles for Agno during the viral infection cycle.JC virus agnoprotein plays critical regulatory roles during the viral replication cycle. Previously, the release of agnoprotein from agnoprotein‐positive cells was reported. In this work, we have further mapped the region(s) responsible for its release. Data showed that the specific amino acid residues located on the hydrophilic surface (designated as release surface) of the major alpha helix domain of agnoprotein, including Lys22, Lys23, Phe31, Glu34, and Asp38 play important roles in this process. Additionally, agnoprotein was found to strongly interact with the unidentified components of the cell surface, suggesting additional roles for it during the viral replication cycle.
      PubDate: 2017-08-28T05:45:56.648492-05:
      DOI: 10.1002/jcp.26106
  • 12‐O‐tetradecanoylphorbol‐13‐acetate and EZH2 inhibition: A novel
           approach for promoting myogenic differentiation in embryonal
           rhabdomyosarcoma cells
    • Authors: Irene Marchesi; Luca Sanna, Milena Fais, Francesco P. Fiorentino, Antonio Giordano, Luigi Bagella
      First page: 2360
      Abstract: Rhabdomyosarcoma (RMS) is a soft tissue sarcoma that arises from muscle precursors affecting predominately children and young adults. It can be divided into two main classes: embryonal (eRMS) and alveolar rhabodomyosarcomas (aRMS). Despite the expression of early muscle specific genes, RMS cells fail to complete myogenesis even in differentiation conditions. We previously demonstrated that Enhancer Zeste of Homolog 2 (EZH2), the catalytic subunits of PRC2 complex, contributes to inhibit muscle differentiation in eRMS and its down‐regulation causes a partial recovery of myogenesis. 12‐O‐tetradecanoylphorbol‐13‐acetate (TPA) is a molecule able to induce differentiation in eRMS with a mechanism that involves the protein kinase C (PKC). In this paper we report that treatment with TPA reduces the expression of EZH2 without affecting levels of H3K27me3. The combination of TPA with GSK126, an inhibitor of the catalytic activity of EZH2, has a synergic effect on the induction of muscle differentiation in RD rhabdomyosarcoma cells, suggesting a new therapeutic combinatory approach for RMS treatment.Rhabdomyosarcoma (RMS) is a soft tissue sarcoma that arises from muscle precursors. We previously demonstrated that down‐regulation of EZH2 in RMS cells causes a partial recovery of myogenesis. We report that the combination of TPA with GSK126 scores a synergic effect on the induction of muscle differentiation, suggesting a new therapeutic combinatory approach for RMS treatment
      PubDate: 2017-08-30T05:40:28.530679-05:
      DOI: 10.1002/jcp.26107
  • The ubiquitin ligase SCFFBXW7α promotes GATA3 degradation
    • Authors: Nan Song; Cheng Cao, Yiman Tang, Liyuan Bi, Yong Jiang, Yongsheng Zhou, Xin Song, Ling Liu, Wenshu Ge
      First page: 2366
      Abstract: GATA3 is a key transcription factor in cell fate determination and its dysregulation has been implicated in various types of malignancies. However, how the abundance and function of GATA3 are regulated remains unclear. Here, we report that GATA3 is physically associated with FBXW7α, and FBXW7α destabilizes GATA3 through assembly of a SKP1‐CUL1‐F‐box E3 ligase complex. Importantly, we showed that FBXW7α promotes GATA3 ubiquitination and degradation in a GSK3 dependent manner. Furthermore, we demonstrated that FBXW7α inhibits breast cancer cells survival through destabilizing GATA3, and the expression level of FBXW7α is negatively correlated with that of GATA3 in breast cancer samples. This study indicated that FBXW7α is a critical negative regulator of GATA3 and revealed a pathway for the maintenance of GATA3 abundance in breast cancer cells.In this study, we aimed to understand how the abundance and function of transcription factor GATA3 are regulated. We revealed that FBXW7α interacts with and promotes turnover of GATA3, and FBXW7α suppresses breast cancer cell survival by degrading GATA3.
      PubDate: 2017-09-04T03:21:21.420346-05:
      DOI: 10.1002/jcp.26108
  • Electric fields accelerate cell polarization and bypass myosin action in
           motility initiation
    • Authors: Yao-Hui Sun; Yuxin Sun, Kan Zhu, Brian Reid, Xing Gao, Bruce W. Draper, Min Zhao, Alex Mogilner
      First page: 2378
      Abstract: Stationary symmetrical fish keratocyte cells break symmetry and become motile spontaneously but slowly. We found that applying electric field (EF) accelerates the polarization by an order of magnitude. While spontaneously polarized cells move persistently for hours, the EF‐induced polarity is lost in a majority of cells when the EF is switched off. However, if the EF is applied for a long time and then switched off, the majority of cell move stably. Myosin inhibition abolishes spontaneous polarization, but does not slow down EF‐induced polarization, and after the EF is turned off, motility does not stop; however, the cell movements are erratic. Our results suggest that the EF rapidly polarizes the cells, but that resulting polarization becomes stable slowly, and that the EF bypasses the requirement for myosin action in motility initiation.Electric field rapidly polarizes fish keratocyte cells, but the resulting polarization becomes stable slowly. Electric field bypasses the requirement for myosin action in motility initiation.
      PubDate: 2017-08-25T05:50:49.513692-05:
      DOI: 10.1002/jcp.26109
  • Mineral trioxide aggregate enhances the osteogenic capacity of periodontal
           ligament stem cells via NF‐κB and MAPK signaling pathways
    • Authors: Yanqiu Wang; Yixiang Zhou, Lin Jin, Xiyao Pang, Yadie Lu, Zilu Wang, Yan Yu, Jinhua Yu
      First page: 2386
      Abstract: Mineral trioxide aggregate (MTA), as a bioactive material, has a widespread application in clinical practice. To date, the effects of MTA on the proliferation and differentiation of human periodontal ligament stem cells (hPDLSCs) remain unclear. hPDLSCs were isolated from human periodontal ligament tissues and cultured with MTA conditioned media. Cell counting kit‐8 (CCK‐8) assay was performed to assess the proliferation capacity of MTA‐treated hPDLSCs. Immunofluorescence assay, alkaline phosphatase (ALP) activity, alizarin red staining, real‐time RT‐PCR, and western blot analyses were used to investigate the odonto/osteogenic capacity of hPDLSCs as well as the involvement of NF‐κB and MAPK pathways. ALP activity assay revealed that 2 mg/ml was the optimal concentration for the induction of hPDLSCs by MTA. The protein expression of DSP, RUNX2, OCN, OSX, OPN, DMP1, ALP, and COL‐I in MTA‐treated hPDLSCs was significantly higher than those in control group (p 
      PubDate: 2017-08-25T05:55:27.885799-05:
      DOI: 10.1002/jcp.26110
  • GSK-3β Inhibition Suppresses Instability-induced Osteolysis by a Dual
           Action on Osteoblast and Osteoclast Differentiation
    • Authors: Mehdi Amirhosseini; Rune V. Madsen, K. Jane Escott, Mathias P. Bostrom, F. Patrick Ross, Anna Fahlgren
      First page: 2398
      Abstract: Currently, there are no medications available to treat aseptic loosening of orthopedic implants. Using osteoprotegerin fusion protein (OPG-Fc), we previously blocked instability-induced osteoclast differentiation and peri-prosthetic osteolysis. Wnt/β-catenin signaling, which regulates OPG secretion from osteoblasts, also modulates the bone tissue response to mechanical loading. We hypothesized that activating Wnt/β–catenin signaling by inhibiting glycogen synthase kinase-3β (GSK-3β) would reduce instability-induced bone loss through regulation of both osteoblast and osteoclast differentiation. We examined effects of GSK-3β inhibition on regulation of RANKL and OPG in a rat model of mechanical instability-induced peri-implant osteolysis. The rats were treated daily with a GSK-3β inhibitor, AR28 (20 mg/kg bw), for up to 5 days. Bone tissue and blood serum were assessed by qRT-PCR, immunohistochemistry and ELISA on days 3 and 5, and by micro-CT on day 5. After 3 days of treatment with AR28, mRNA levels of β-catenin, Runx2, Osterix, Col1α1 and ALP were increased leading to higher osteoblast numbers compared to vehicle-treated animals. BMP-2 and Wnt16 mRNA levels were downregulated by mechanical instability and this was rescued by GSK-3β inhibition. Osteoclast numbers were decreased significantly after 3 days of GSK-3β inhibition, which correlated with enhanced OPG mRNA expression. This was accompanied by decreased serum levels of TRAP5b on days 3 and 5. Treatment with AR28 upregulated osteoblast differentiation, while osteoclastogenesis was blunted, leading to increased bone mass by day 5. These data suggest that GSK-3β inactivation suppresses osteolysis through regulating both osteoblast and osteoclast differentiation in a rat model of instability-induced osteolysis. This article is protected by copyright. All rights reserved
      PubDate: 2017-09-28T04:01:50.20423-05:0
      DOI: 10.1002/jcp.26111
  • TNF‐α‐induced NF‐κB activation promotes myofibroblast
           differentiation of LR‐MSCs and exacerbates bleomycin‐induced pulmonary
    • Authors: Jiwei Hou; Tan Ma, Honghui Cao, Yabing Chen, Cong Wang, Xiang Chen, Zou Xiang, Xiaodong Han
      First page: 2409
      Abstract: Idiopathic pulmonary fibrosis (IPF) is a chronic, progressive, and irreversible lung disease of unknown cause. It has been reported that both lung resident mesenchymal stem cells (LR‐MSCs) and tumor necrosis factor‐α (TNF‐α) play important roles in the development of pulmonary fibrosis. However, the underlying connections between LR‐MSCs and TNF‐α in the pathogenesis of pulmonary fibrosis are still elusive. In this study, we found that the pro‐inflammatory cytokine TNF‐α and the transcription factor nuclear factor kappa B (NF‐κB) p65 subunit were both upregulated in bleomycin‐induced fibrotic lung tissue. In addition, we discovered that TNF‐α promotes myofibroblast differentiation of LR‐MSCs through activating NF‐κB signaling. Interestingly, we also found that TNF‐α promotes the expression of β‐catenin. Moreover, we demonstrated that suppression of the NF‐κB signaling could attenuate myofibroblast differentiation of LR‐MSCs and bleomycin‐induced pulmonary fibrosis which were accompanied with decreased expression of β‐catenin. Our data implicates that inhibition of the NF‐κB signaling pathway may provide a therapeutic strategy for pulmonary fibrosis, a disease that warrants more effective treatment approaches.TNF‐α promotes myofibroblast differentiation of LR‐MSCs and pulmonary fibrosis through activating the NF‐κB signaling pathway.
      PubDate: 2017-08-25T05:36:19.181249-05:
      DOI: 10.1002/jcp.26112
  • The ubiquitin ligase TRIM56 inhibits ovarian cancer progression by
           targeting vimentin
    • Authors: Lei Zhao; Ping Zhang, Xiao-jie Su, Bing Zhang
      First page: 2420
      Abstract: Tumor metastasis is responsible for 90% of all cancer‐related deaths. Epithelial to mesenchymal transition (EMT) is an important prerequisite for tumor metastasis. One of the important mediators of EMT and cancer progression in ovarian cancer is the vimentin protein. The objective of the current study was to evaluate the molecular mechanism that regulates vimentin expression in ovarian cancer cells. Vimentin was robustly induced in the ovarian cancer cell line SKOV‐3 compared to normal ovarian epithelial cell line Moody and the induction was not due to transcriptional upregulation. Treatment with the proteasomal inhibitor MG‐132 revealed that vimentin is actively degraded by the proteasome in Moody cells and stabilized in the SKOV‐3 cell line. Mass spectrometric analysis of vimentin immunoprecipitate of MG‐132 treated Moody cells revealed candidate ubiquitin ligases associated with vimentin. RNAi mediated silencing of the candidate ubiquitin in Moody cells and concurrent overexpression of the candidate ubiquitin ligases in SKOV‐3 confirmed that TRIM56 is the ubiquitin ligase that is degrading vimentin in Moody cells. RNAi mediated silencing of TRIM56 in Moody cells and ectopic overexpression of TRIM56 in SKOV‐3 cells, respectively, significantly up‐ and down‐regulated in vitro migration and invasion in these cells. Analysis of TRIM56 transcript level and vimentin protein expression in 25 patients with ovarian carcinoma confirmed an inverse correlation between TRIM56 and vimentin expression. Cumulatively, our data reveals for the first time a novel post‐translational regulatory mechanism of regulating vimentin expression, EMT, and metastatic progression in ovarian cancer cells.RNAi mediated silencing of TRIM56 in Moody cells and ectopic overexpression of TRIM56 in SKOV‐3 cells, respectively, significantly up‐ and down‐regulated in vitro migration and invasion in these cells. Analysis of TRIM56 transcript level and vimentin protein expression in 25 patients with ovarian carcinoma confirmed an inverse correlation between TRIM56 and vimentin expression.
      PubDate: 2017-09-12T06:25:23.745599-05:
      DOI: 10.1002/jcp.26114
  • EPA blocks TNF‐α‐induced inhibition of sugar uptake in Caco‐2 cells
           via GPR120 and AMPK
    • Authors: Rosa Castilla-Madrigal; Jaione Barrenetxe, María J. Moreno-Aliaga, María Pilar Lostao
      First page: 2426
      Abstract: The aim of the present work was to investigate in Caco‐2 cells whether eicosapentaenoic acid (EPA), an omega‐3 polyunsaturated fatty acid, could block the inhibitory effect of tumor necrosis factor‐α (TNF‐α) on sugar transport, and identify the intracellular signaling pathways involved. After pre‐incubation of the Caco‐2 cells with TNF‐α and EPA for 1 hr, EPA prevented the inhibitory effect of the cytokine on α‐methyl‐d‐glucose (αMG) uptake (15 min) and on SGLT1 expression at the brush border membrane, measured by Western blot. The ERK1/2 inhibitor PD98059 and the AMPK activator AICAR also prevented the inhibitory effect of TNF‐α on both αMG uptake and SGLT1 expression. Interestingly, the AMPK inhibitor, Compound C, abolished the ability of EPA to prevent TNF‐α‐induced reduction of sugar uptake and transporter expression. The GPR120 antagonist, AH7614, also blocked the preventive effect of EPA on TNF‐α‐induced decrease of αMG uptake and AMPK phosphorylation. In summary, TNF‐α inhibits αMG uptake by decreasing SGLT1 expression in the brush border membrane through the activation of ERK1/2 pathway. EPA prevents the inhibitory effect of TNF‐α through the involvement of GPR120 and AMPK activation.TNF‐α decreases SGLT1 expression in the plasma membrane by activation of ERK and inhibition of AMPK pathways. EPA prevents TNF‐α decrease of sugar uptake by activating AMPK pathways. EPA blocks TNF‐α effect on sugar uptake through GPR120.
      PubDate: 2017-08-30T05:28:04.752843-05:
      DOI: 10.1002/jcp.26115
  • β‐asarone inhibited cell growth and promoted autophagy via
           P53/Bcl‐2/Bclin‐1 and P53/AMPK/mTOR pathways in Human Glioma U251
    • Authors: Nanbu Wang; Qinxin Zhang, Laiyu Luo, Baile Ning, Yongqi Fang
      First page: 2434
      Abstract: Glioma is the most common type of primary brain tumor and has an undesirable prognosis. Autophagy plays an important role in cancer therapy, but it is effect is still not definite. P53 is an important tumor suppressor gene and protein that is closely to autophagy. Our aim was to study the effect of β‐asarone on inhibiting cell proliferation in human glioma U251 cells and to detect the effect of the inhibition on autophagy through the P53 signal pathway. For cell growth, the cells were divided into four groups: the model, β‐asarone, temozolomide (TMZ), and co‐administration groups. For cell autoghapy and the P53 pathway, the cells were divided into six groups: the model, β‐asarone, 3MA, Rapa, Pifithrin‐µ, and NSC groups. The counting Kit‐8 assay and flow cytometry (FCM) were then used to measure the cell proliferation and cycle. Electron microscopy was used to observe autophagosome formation. Cell immunohistochemistry/‐immunofluorescence, FCM and Western blot (WB) were used to examine the expression of Beclin‐1 and P53. The levels of P53 and GAPDH mRNA were detected by RT‐PCR. Using WB, we determined autophagy‐related proteins Beclin‐1, LC3‐II/I, and P62 and those of the P53 pathway‐related proteins P53, Bcl‐2, mTOR, P‐mTOR, AMPK, P‐AMPK, and GAPDH. We got the results that β‐asarone changed the cellular morphology, inhibited cell proliferation, and enhanced the expression of P53, LC3‐II/I, Beclin‐1, AMPK, and pAMPK while inhibiting the expression of P62, Bcl‐2, mTOR, and pmTOR. All the data suggested that β‐asarone could reduce the cell proliferation and promote autophagy possible via the P53 pathway in U251 cells.β‐asarone inhibits the growth of glioma U251 cells. β‐asarone promotes autophagy of U251 cells. The autophagy process possible be promoted through the P53/Bcl‐2/Bclin‐1 and P53/AMPK/mTOR signal pathway.
      PubDate: 2017-08-30T05:30:40.236912-05:
      DOI: 10.1002/jcp.26118
  • Hypomethylation‐mediated H19 overexpression increases the risk of
           disease evolution through the association with BCR‐ABL transcript in
           chronic myeloid leukemia
    • Authors: Jing-dong Zhou; Jiang Lin, Ting-juan Zhang, Ji-chun Ma, Xi-xi Li, Xiang-mei Wen, Hong Guo, Zi-jun Xu, Zhao-qun Deng, Wei Zhang, Jun Qian
      First page: 2444
      Abstract: Previous study has revealed that H19 expression is required for efficient tumor growth induced by BCR‐ABL in chronic myeloid leukemia (CML). Herein, we further determined H19 expression and its clinical implication in patients with CML. H19 expression and methylation were detected by real‐time quantitative PCR and real‐time quantitative methylation‐specific PCR, and then clinical implication of H19 expression was further analyzed. H19 expression was significantly up‐regulated in CML patients (p 
      PubDate: 2017-08-30T05:35:50.270751-05:
      DOI: 10.1002/jcp.26119
  • Potential role of the Jagged1/Notch1 signaling pathway in the
           endothelial‐myofibroblast transition during BLM‐induced pulmonary
    • Authors: Qian Yin; Weihua Wang, Guangbin Cui, Linfeng Yan, Song Zhang
      First page: 2451
      Abstract: Endothelial cell myofibroblast transition (EndoMT) is found during the process of bleomycin (BLM)‐induced pulmonary fibrosis in rats, and plays a very important role in sustaining inflammation and collagen secretion. Moreover, some studies have suggested that the Notch1 signaling pathway may be involved in the expression of α‐smooth muscle actin (α‐SMA) in pulmonary microvascular endothelial cells (PMVECs), a protein marker of EndoMT. Therefore, we aimed to investigate the expression level of α‐SMA and Notch1‐related signaling molecules in PMVECs from BLM‐induced rats and determine the relationship between the Notch1 signaling pathway and the expression of α‐SMA in PMVECs. We found that the expression levels of α‐SMA, Notch1, and Jagged1 were upregulated, while the expression levels of Dll4 were downregulated. Furthermore, there was a positive correlation between the expression of Jagged1 and the α‐SMA proteins in PMVECs, and NF‐κB was downregulated by decreasing the expression of Jagged1. In conclusion, the Jagged1/Notch1 signaling pathway is activated in PMVECs during the pathogenesis of BLM‐induced pulmonary fibrosis in rats, and it may induce α‐SMA expression via a non‐canonical pathway involving NF‐κB as the target molecule. The precise mechanism and the molecules involved in this signaling pathway need to be further elucidated.The Jagged1/Notch1 signaling pathway is activated in PMVECs during the pathogenesis of BLM‐induced pulmonary fibrosis in rats, and it may induce α‐SMA expression via a non‐canonical pathway involving NF‐κB as the target molecule.
      PubDate: 2017-09-07T04:10:45.199681-05:
      DOI: 10.1002/jcp.26122
  • Annexin A2 positively regulates milk synthesis and proliferation of bovine
           mammary epithelial cells through the mTOR signaling pathway
    • Authors: Minghui Zhang; Dongying Chen, Zhen Zhen, Jinxia Ao, Xiaohan Yuan, Xuejun Gao
      First page: 2464
      Abstract: Annexin A2 (AnxA2) has been shown to play multiple roles in growth, development, and metabolism, but the functions of AnxA2 and the signaling pathways associated with AnxA2 are still not fully understood. In this study, we aim to reveal whether and how AnxA2 could be involved in milk synthesis and proliferation of bovine mammary epithelial cells (BMECs). Using gene function study approaches, we found that AnxA2 positively regulates PIP3 level, phosphorylation of mTOR, and protein levels of SREBP‐1c and Cyclin D1 leading to milk synthesis and cell proliferation. We further observed that both AnxA2‐36 kD phosphorylated form and AnxA2‐33 kD protein could be induced from AnxA2‐36 kD protein in BMECs under methionine, leucine, estrogen or prolactin stimulation. These above results strongly demonstrate that AnxA2 functions as a critical regulator for amino acid or hormone‐induced milk synthesis and cell proliferation via the PI3K‐mTOR‐SREBP‐1c/Cyclin D1 signaling pathway.Annexin A2 is phosphorylated and further processed into a truncated form in response to Met, Leu, E and PRL stimulation, and positively regulates milk synthesis and proliferation of bovine mammary epithelial cells through the mTOR signaling pathway
      PubDate: 2017-08-30T05:26:40.655679-05:
      DOI: 10.1002/jcp.26123
  • MiR‐146a protects small intestine against ischemia/reperfusion injury by
           down‐regulating TLR4/TRAF6/NF‐κB pathway
    • Authors: Xuemei He; Yingqiang Zheng, Shengzhi Liu, Sen Shi, Yong Liu, Yanzheng He, Chunxiang Zhang, Xiangyu Zhou
      First page: 2476
      Abstract: Previous studies reported that miR‐146a was involved in small intestine ischemia‐reperfusion (I/R) injury, but the mechanism is largely vague. Here, we aimed to identify the change of miR‐146a in patients with mesenteric ischemia and explore the potential regulatory mechanism of miR‐146a in intestine epithelial cells survival under ischemia and I/R injury. The plasma of 20 patients with mesenteric ischemia and 25 controls was collected to examine the miR‐146a expression by qPCR. Rat intestinal epithelial cells (IEC‐6) and 24 male Sprague–Dawley rats were included to build ischemia and I/R model in vitro and in vivo. The qPCR results showed that miR‐146a decreased both in the plasma of patients with mesenteric ischemia and in IEC‐6 cells and rat small intestine tissues in ischemia and I/R model compared to controls. Both the in vitro and in vivo results showed that I/R resulted in more severe apoptotic injury than ischemia. Cleaved‐caspase 3, TLR4, TRAF6, and nuclear NF‐κB p65 were up‐regulated accompanying reduced XIAP and SOCS3 expression in intestinal ischemia and I/R injury. After up‐regulation of miR‐146a in IEC‐6 cells, increased cell survival and decreased cell apoptosis were observed, concomitant with decreased cleaved‐caspase 3 and down‐regulated TLR4/TRAF6/NF‐κB pathway. What is more, this protective effect was blocked by TRAF6 overexpression and increased nuclear NF‐κB p65 nuclear. Taken together, this study revealed that miR‐146a expression was decreased in small intestine ischemia and I/R injury. And miR‐146a improves intestine epithelial cells survival under ischemia and I/R injury through inhibition TLR4, TRAF6, and p‐IκBα, subsequently leading to decreased NF‐κB p65 nuclear translocation.miR‐146a expression was decreased in small intestine ischemia and I/R injury. And miR‐146a improves intestine epithelial cells survival under ischemia and I/R injury through inhibition TLR4 and TRAF6, subsequently leading to decreased NF‐κB p65 nuclear translocation.
      PubDate: 2017-08-25T05:40:39.01312-05:0
      DOI: 10.1002/jcp.26124
  • TLR4 and NFκB signaling is critical for taxol resistance in ovarian
           carcinoma cells
    • Authors: Nian-Kang Sun; Shang-Lang Huang, Ting-Chang Chang, Chuck C.-K. Chao
      First page: 2489
      Abstract: We report here that toll‐like receptor 4 (TLR4) and ABCB1 are upregulated in SKOV3 ovarian carcinoma cells that acquired resistance to the anticancer drug taxol. Silencing of TLR4 using short‐hairpin RNA sensitized taxol‐resistant SKOV3 cells to taxol (4.6 fold), whereas ectopic expression of TLR4 in parental, taxol‐sensitive SKOV3 cells or TLR4‐null HEK293 cells induced taxol resistance (∼2 fold). A sub‐lethal dose of taxol induced ABCB1 protein expression in taxol‐resistant SKOV3 cells. Inactivation of TLR4 using chemical inhibitors (CLI‐095 and AO‐I) downregulated ABCB1 protein expression and enhanced the cytotoxic activity of taxol in taxol‐resistant SKOV3 cells. While the sensitization effect of TLR4 inactivation was also detected in TOV21G ovarian cancer cells, which express moderate level of TLR4, ectopic expression of ABCB1 prevented the sensitization effect in these cells. Notably, the NFκB pathway was significantly activated by taxol, and inhibition of this pathway suppressed TLR4‐regulated ABCB1 expression. Furthermore, taxol‐induced NFκB signaling was reduced following TLR4 silencing in taxol‐resistant SKOV3 cells. Consistent with these results, ectopic expression of TLR4 in taxol‐sensitive SKOV3 cells enhanced ABCB1 expression and conferred resistance to taxol. The protective effect of exogenous TLR4 expression against taxol was reduced by treatment with NFκB inhibitor in these cells. These results demonstrate that taxol activates the TLR4‐NFκB pathway which in turn induces ABCB1 gene expression. This cellular pathway thus represents a novel target to limit resistance to taxol in ovarian cancer cells.The taxol activates the TLR4‐NFkB pathway which in turn induces ABCB1 gene expression. This cellular pathway thus represents a novel target to limit resistance to taxol in ovarian cancer cells.
      PubDate: 2017-08-30T05:31:35.282745-05:
      DOI: 10.1002/jcp.26125
  • Cyanidin Chloride inhibits ovariectomy‐induced osteoporosis by
           suppressing RANKL‐mediated osteoclastogenesis and associated signaling
    • Authors: Jianwen Cheng; Lin Zhou, Qian Liu, Jennifer Tickner, Zhen Tan, Xiaofeng Li, Mei Liu, Xixi Lin, Tao Wang, Nathan J. Pavlos, Jinmin Zhao, Jiake Xu
      First page: 2502
      Abstract: Over‐production and activation of osteoclasts is a common feature of osteolytic conditions such as osteoporosis, tumor‐associated osteolysis, and inflammatory bone erosion. Cyanidin Chloride, a subclass of anthocyanin, displays antioxidant and anti‐carcinogenesis properties, but its role in osteoclastic bone resorption and osteoporosis is not well understood. In this study, we showed that Cyanidin Chloride inhibits osteoclast formation, hydroxyapatite resorption, and receptor activator of NF‐κB ligand (RANKL)‐induced osteoclast marker gene expression; including ctr, ctsk, and trap. Further investigation revealed that Cyanidin Chloride inhibits RANKL‐induced NF‐κB activation, suppresses the degradation of IκB‐α and attenuates the phosphorylation of extracellular signal‐regulated kinases (ERK). In addition, Cyanidin Chloride abrogated RANKL‐induced calcium oscillations, the activation of nuclear factor of activated T cells calcineurin‐dependent 1 (NFATc1), and the expression of c‐Fos. Further, we showed that Cyanidin Chloride protects against ovariectomy‐induced bone loss in vivo. Together our findings suggest that Cyanidin Chloride is capable of inhibiting osteoclast formation, hydroxyapatite resorption and RANKL‐induced signal pathways in vitro and OVX‐induced bone loss in vivo, and thus might have therapeutic potential for osteolytic diseases.Cyanidin Chloride inhibits osteoclast formation, hydroxyapatite resorption, and receptor activator of NF‐kB ligand (RANKL)‐induced osteoclast marker gene expression; including ctr, ctsk, and trap. Cyanidin Chloride inhibits RANKL‐induced NF‐kB activation, suppresses the degradation of IkB‐a and attenuates the phosphorylation of extracellular signal‐regulated kinases (ERK). Cyanidin Chloride also abrogates RANKL‐induced calcium oscillations, the activation of nuclear factor of activated T cells calcineurin‐dependent 1 (NFATc1), and the expression of c‐Fos. Cyanidin Chloride protects against ovariectomy‐induced bone loss in vivo. Cyanidin Chloride is capable of inhibiting osteoclast formation, hydroxyapatite resorption and RANKL‐induced signal pathways in vitro and OVX‐induced bone loss in vivo, and thus might have therapeutic potential for osteolytic diseases.
      PubDate: 2017-09-07T04:17:35.198481-05:
      DOI: 10.1002/jcp.26126
  • Tannic acid attenuates TGF‐β1‐induced epithelial‐to‐mesenchymal
           transition by effectively intervening TGF‐β signaling in lung
           epithelial cells
    • Authors: Dhamotharan Pattarayan; Ayyanar Sivanantham, Venkateshwaran Krishnaswami, Lakshmanan Loganathan, Rajaguru Palanichamy, Subramanian Natesan, Karthikeyan Muthusamy, Subbiah Rajasekaran
      First page: 2513
      Abstract: Idiopathic pulmonary fibrosis (IPF) is a chronic, progressive, and an irreversible lung disorder characterized by the accumulation of fibroblasts and myofibroblasts in the extracellular matrix. The transforming growth factor‐β1 (TGF‐β1)‐induced epithelial‐to‐mesenchymal transition (EMT) is thought to be one of the possible sources for a substantial increase in the number of fibroblasts/myofibroblasts in IPF lungs. Tannic acid (TA), a natural dietary polyphenolic compound has been shown to possess diverse pharmacological effects. However, whether TA can inhibit TGF‐β1‐mediated EMT in lung epithelial cells remains enigmatic. Both the human adenocarcinomic alveolar epithelial (A549) and normal bronchial epithelial (BEAS‐2B) cells were treated with TGF‐β1 with or without TA. Results showed that TA addition, markedly inhibited TGF‐β1‐induced EMT as assessed by reduced expression of N‐cadherin, type‐1‐collagen, fibronectin, and vimentin. Furthermore, TA inhibited TGF‐β1‐induced cell proliferation through inducing cell cycle arrest at G0/G1 phase. TGF‐β1‐induced increase in the phosphorylation of Smad (Smad2 and 3), Akt as well as that of mitogen activated protein kinase (ERK1/2, JNK1/2, and p38) mediators was effectively inhibited by TA. On the other hand, TA reduced the TGF‐β1‐induced increase in TGF‐β receptors expression. Using molecular docking approach, FTIR, HPLC and Western blot analyses, we further identified the direct binding of TA to TGF‐β1. Finally, we conclude that TA might directly interact with TGF‐β1, thereby repressing TGF‐β signaling and subsequent EMT process in lung epithelial cells. Further animal studies are needed to clarify its potential therapeutic benefit in pulmonary fibrosis.Tannic acid suppresses TGF‐β1‐induced EMT.
      PubDate: 2017-08-30T05:20:41.956496-05:
      DOI: 10.1002/jcp.26127
  • Inhibition of ZL55 cell proliferation by ADP via PKC‐dependent
           signalling pathway
    • Authors: Antonella Muscella; Luca G. Cossa, Carla Vetrugno, Giovanna Antonaci, Santo Marsigliante
      First page: 2526
      Abstract: Extracellular nucleotides can regulate cell proliferation in both normal and tumorigenic tissues. Here, we studied how extracellular nucleotides regulate the proliferation of ZL55 cells, a mesothelioma‐derived cell line obtained from bioptic samples of asbestos‐exposed patients. ADP and 2‐MeS‐ADP inhibited ZL55 cell proliferation, whereas ATP, UTP, and UDP were inactive. The nucleotide potency profile and the blockade of the ADP‐mediated inhibitory effect by the phospholipase C inhibitor U‐73122 suggest that P2Y1 receptor controls ZL55 cell proliferation. The activation of P2Y1 receptor by ADP leads to activation of intracellular transduction pathways involving [Ca2+]i, PKC‐δ/PKC‐α, and MAPKs, ERK1/2 and JNK1/2. Cell treatment with ADP or 2‐MeS‐ADP also provokes the activation of p53, causing an accumulation of the G1 cyclin‐dependent kinase inhibitors p21WAF1 and p27Kip. Inhibition of ZL55 cell proliferation by ADP was completely reversed by inhibiting MEK1/2, or JNK1/2, or PKC‐δ, and PKC‐α. Through the inhibition of ADP‐activated transductional kinases it was found that PKC‐δ was responsible for JNK1/2 activation. JNK1/2 has a role in transcriptional up‐regulation of p53, p21WAF1/CIP1, and p27kip1. Conversely, the ADP‐activated PKC‐α provoked ERK1/2 phosphorylation. ERK1/2 increased p53 stabilization, required to G1 arrest of ZL55 cells. Concluding, the importance of the study is twofold: first, results shed light on the mechanism of cell cycle inhibition by ADP; second, results suggest that extracellular ADP may inhibit mesothelioma progression.In human mesothelioma ZL55 cells, activation of P2Y1 receptor by ADP leads to activation of intracellular transduction pathways involving [Ca2+]i, PKC‐δ/PKC‐α, and MAPKs ERK1/2 and JNK1/2. Cell treatment with ADP or 2‐MeS‐ADP also provokes the activation of p53, causing an accumulation of the G1 cyclin‐dependent kinase inhibitors p21WAF1 and p27Kip. ADP‐activated PKC‐δ was responsible of JNK1/2 activation that has a role in transcriptional up‐regulation of p53, p21WAF1, and p27kip1. Conversely, the ADP‐activated PKC‐α provoked ERK1/2 phosphorylation that increased p53 stabilization, required to G1 arrest of ZL55 cells. Concluding, the importance of the study is twofold: first, results shed light on the mechanism of cell cycle inhibition by ADP; second, results suggest that extracellular ADP may inhibit mesothelioma progression.
      PubDate: 2017-09-04T03:26:53.685156-05:
      DOI: 10.1002/jcp.26128
  • Double sex and mab‐3 related transcription factor 1 regulates
           differentiation and proliferation in dairy goat male germline stem cells
    • Authors: Yudong Wei; Shufang Cai, Fanglin Ma, Ying Zhang, Zhe Zhou, Shuanshuan Xu, Mengfei Zhang, Sha Peng, Jinlian Hua
      First page: 2537
      Abstract: The protein encoded by double sex and mab‐3 related transcription factor 1 (Dmrt1) gene contains a double sex/mab‐3 domain, which was considered as one of the most conservative structures in sex determination. However, its effect on spermatogenesis of dairy goat spermatogonial stem cells (SSCs) remains to be clarified. For the first time, the roles of Dmrt1 in spermatogenesis of livestock are highlighted. Here, we investigated the expression pattern of Dmrt1 in the testes of dairy goats. Dmrt1 primarily located in undifferentiated SSCs. Moreover, Dmrt1 enhanced differentiation and proliferation of mGSCs. On the contrary, the level of meiosis was down‐regulated, as Dmrt1 determines whether SSCs undergo mitosis and spermatogonial differentiation or meiosis. In the busulfan‐treated mice testes, Dmrt1 repair germ cell damage was emphasized as well. Our results exposed that Dmrt1 maintenance mGSCs in two ways: facilitating proliferation and self‐renewal of SSCs; and reducing the inflammatory response caused by reproductive injury. These findings identify a central role for Dmrt1 in controlling population stability and injury restoring of SSCs.For the first time, the roles of Dmrt1 in spermatogenesis of livestock are highlighted. Dmrt1 primarily located in undifferentiated SSCs, moreover, Dmrt1 enhanced differentiation and proliferation of mGSCs. Our results exposed that Dmrt1 maintenance mGSCs in two ways: facilitating proliferation and self‐renewal of SSCs; and reducing the inflammatory response caused by reproductive injury.
      PubDate: 2017-09-13T05:41:57.291621-05:
      DOI: 10.1002/jcp.26129
  • Spleen tyrosine kinase influences the early stages of multilineage
           differentiation of bone marrow stromal cell lines by regulating
           phospholipase C gamma activities
    • Authors: Joji Kusuyama; Ai Kamisono, Seong ChangHwan, Muhammad S. Amir, Kenjiro Bandow, Nahoko Eiraku, Tomokazu Ohnishi, Tetsuya Matsuguchi
      First page: 2549
      Abstract: Bone marrow stromal cells (BMSCs) are multipotent cells that can differentiate into adipocytes and osteoblasts. Inadequate BMSC differentiation is occasionally implicated in chronic bone metabolic disorders. However, specific signaling pathways directing BMSC differentiation have not been elucidated. Here, we explored the roles of spleen tyrosine kinase (Syk) in BMSC differentiation into adipocytes and osteoblasts. We found that Syk phosphorylation was increased in the early stage, whereas its protein expression was gradually decreased during the adipogenic and osteogenic differentiation of two mouse mesenchymal stromal cell lines, ST2 and 10T(1/2), and a human BMSC line, UE6E-7-16. Syk inactivation with either a pharmacological inhibitor or Syk-specific siRNA suppressed adipogenic differentiation, characterized by decreased lipid droplet appearance and the gene expression of fatty acid protein 4 (Fabp4), peroxisome proliferator-activated receptor γ2 (Pparg2), CCAAT/enhancer binding proteins α(C/EBPα) and C/EBPβ . In contrast, Syk inhibition promoted osteogenic differentiation, represented by increase in matrix mineralization and alkaline phosphatase (ALP) activity, as well as the expression levels of osteocalcin, runt-related transcription factor 2 (Runx2) and distal-less homeobox 5 (Dlx5) mRNAs. We also found that Syk-induced signals are mediated by phospholipase Cγ1 (PLCγ1) in osteogenesis and PLCγ2 in adipogenesis. Notably, Syk-activated PLC 2 signaling was partly modulated through B-cell linker protein(BLNK) in adipogenic differentiation. On the other hand, growth factor receptor-binding protein 2 (Grb2) was involved in Syk-PLCγ1 axis in osteogenic differentiation. Taken together, these results indicate that Syk-PLCγ signaling has a dual role in regulating the initial stage of adipogenic and osteogenic differentiation of BMSCs. This article is protected by copyright. All rights reserved
      PubDate: 2017-09-28T03:50:31.105124-05:
      DOI: 10.1002/jcp.26130
  • Bifunctional role of ephrin A1‐Eph system in stimulating cell
           proliferation and protecting cells from cell death through the attenuation
           of ER stress and inflammatory responses in bovine mammary epithelial cells
    • Authors: Minkyung Kang; Wooyoung Jeong, Hyocheol Bae, Whasun Lim, Fuller W. Bazer, Gwonhwa Song
      First page: 2560
      Abstract: Structural and functional development of the mammary gland is constant in the mammary gland life cycle. Eph receptors and their ligands, ephrins, control events through cell‐to‐cell interactions during embryonic development, and adult tissue homeostasis; however, little information on participation of ephrin A1, a representative ligand of the Eph receptor, in the development and function of normal mammary glands is known. In this study, we demonstrated functional effects of the ephrin A1‐Eph system and mechanisms of its action on bovine mammary epithelial (MAC‐T) cells. The in vitro cultured MAC‐T cells expressed the ephrin A1 ligand and EphA1, A2, A4, A7, and A8 among the eight members of the Eph A family. Our results revealed that ephrin A1 induced MAC‐T cell cycle progression and stimulated cell proliferation with abundant expression of nucleic PCNA and cyclin D1 proteins. Additionally, ephrin A1 induced activation of intracellular signaling molecules involved in PI3 K/AKT and MAPK signaling, and the proliferation‐stimulating effect of ephrin A1 was mediated by activation of these pathways. Furthermore, ephrin A1 influenced expression and activation of various ER stress‐related proteins and protected MAC‐T cells from stress‐induced cell death. Finally, ephrin A1 alleviated LPS‐induced cell death through down‐regulation of inflammatory cytokines. In conclusion, the results of this study suggest that the Eph A‐ephrin A1 system is a positive factor in the increase and maintenance of epithelial cells in mammary glands of cows; the signaling system contributes to development, remodeling, and functionality of normal mammary glands and could overcome mastitis in cows and other mammals.The Eph A‐ephrin A1 system is a positive factor in the increase and maintenance of epithelial cells in mammary glands of cows; the signaling system contributes to development, remodeling, and functionality of normal mammary glands and could overcome mastitis in cows and other mammals.
      PubDate: 2017-09-11T09:57:12.366355-05:
      DOI: 10.1002/jcp.26131
  • Iodixanol versus iopromide in cancer patients: Evidence from a randomized
           clinical trial
    • Authors: Irene Terrenato; Francesca Sperati, Felice Musicco, Anna F. Pozzi, Annunziata di Turi, Mauro Caterino, Elisabetta de Lutio di Castelguidone, Sergio V. Setola, Massimo Bellomi, Carlo E. Neumaier, Laura Conti, Giovanni Cigliana, Roberta Merola, Anna Antenucci, Giulia Orlandi, Antonio Giordano, Maddalena Barba, Stefano Canitano
      First page: 2572
      Abstract: To assess the safety profile of iso‐osmolar contrast medium (CM) versus low osmolar CM in cancer patients with an estimated glomerular filtration rate (eGFR) >60 ml/min. In this multicenter, blind trial of patients seeking a chest‐abdomen‐pelvis contrast enhanced computed tomography (CT) with iodated CM, participants were centrally randomized to iodixanol or iopromide. Contrast induced nephropathy (CIN) at 24 and/or 72 hr were our primary outcomes. We further considered irreversible CIN, average eGFR percentage variation (%Δ), and adverse events (AEs). Overall, 607 patients were enrolled. Among them, 497 eligible patients were randomized to iodixanol (N: 247) or iopromide (N: 250). No differences emerged by descriptive characteristics. Seven and 3 CIN at 24 hr (p = 0.34) and 8 and 2 CIN at 72 hr (p = 0.11) occurred in the iopromide and iodixanol group, respectively. Within the subgroup of individual patients who developed CIN (N: 17), the event rate was higher in the iopromide arm (p = 0.045). No cases of permanent CIN or significant differences in terms of AEs or GFR %Δ were observed. Our results suggest a more favorable safety profile of iodixanol versus iopromide. Adequately sized trials with similar design are warranted to confirm our findings and clarify the underlying biological mechanisms.1) This is a multicenter, blind trial of patients seeking a chest‐abdomen‐pelvis CT with iodated CM. 2) Aim of the study was to assess the safety profile of iso‐osmolar contrast medium (CM) versus low osmolar CM in cancer patients with an estimated glomerular filtration rate (eGFR) >60 ml/min. 3) No cases of permanent CIN or significant differences in terms of adverse events were observed.
      PubDate: 2017-09-12T06:10:28.193815-05:
      DOI: 10.1002/jcp.26132
  • Characterization of a novel EB1 acetylation site important for the
           regulation of microtubule dynamics and cargo recruitment
    • Authors: Songbo Xie; Yang Yang, Xiaochen Lin, Jun Zhou, Dengwen Li, Min Liu
      First page: 2581
      Abstract: Microtubule plus ends undergo highly dynamic modifications to regulate different aspects of cellular activities. Most microtubule plus-end tracking proteins (+TIPs) are recruited to the microtubule ends by the master loading factor, end-binding protein 1 (EB1). These proteins coordinately regulate microtubule dynamics and cellular plasticity. Acetylation is known to modulate EB1 function; however, the molecular details of EB1 acetylation remain largely unclear. We mapped the acetylation pattern of EB1 and identified several previously uncharacterized sites of EB1 acetylation. We examined the effects of lysine-212 (K212) acetylation and found that acetylation of this site accelerates autophagy-mediated EB1 degradation. By time-lapse microscopy, we found that the acetylation-deficient K212R mutant increased the percentage of fast-growing and long-lived microtubules. Although K212 acetylation did not affect microtubule stability in vitro and the association of EB1 with microtubules, the K212R mutant significantly promoted microtubule regrowth in cells. Coimmunoprecipitation assays further revealed that the K212 site was critical for the recruitment of different +TIP cargoes. These data thus uncover a critical role for a novel EB1 acetylation site in regulating the dynamic structure of microtubules. This article is protected by copyright. All rights reserved
      PubDate: 2017-09-25T04:50:49.342287-05:
      DOI: 10.1002/jcp.26133
  • Increased expression of PD‐L1 and PD‐L2 in dermal fibroblasts
           from alopecia areata mice
    • Authors: Yunyuan Li; Ruhi T. Kilani, Mohammadreza Pakyari, Gigi Leung, Layla Nabai, Aziz Ghahary
      First page: 2590
      Abstract: Alopecia areata (AA) is a common autoimmune disorder affecting millions of people worldwide, which manifests as a sudden, non‐scarring hair loss. The expression of a pro‐inflammatory cytokine, interferon‐gamma (INF‐γ), has been well established to be involved in the development of AA. As IFN‐γ and other cytokines are also known to up‐regulate programmed cell death ligand 1 and 2 (PD‐L1 and PD‐L2), which both negatively control immune responses, we asked whether or not a high number of infiltrated T cells, seen in AA lesions, can modulate the expression of PD‐L1 and PD‐L2 in skin cells. From a series of experiments, we showed that a significantly higher number of PD‐L1 or PD‐L2 positive cells affect the skin in AA mice, compared to the skin of non‐AA mice. The number of PD‐L1 positive cells was well correlated with the number of infiltrated T cells, especially CD8+ T cells. We also found that the expression of PD‐L1 and PD‐L2 was co‐localized with type 1 pro‐collagen, CD90 and vimentin, which are biomarkers for dermal fibroblasts. Further studies revealed that releasable factors from activated, but not inactivated, lymphocytes significantly increase the expressions of both PD‐L1 and PD‐L2 in cultured dermal fibroblasts. In conclusion, our findings suggest that the expression of PD‐L1 and PD‐L2 in dermal fibroblasts is up‐regulated by activated T cells in AA‐affected skin, and as such, these regulatory molecules may not exert a negative control of the immune activation seen in AA lesions.(i) The highly expression of PD‐L1 and PD‐L2 were found in AA‐affected skin in a mouse mode; (ii) we identified dermal fibroblasts as the skin cells to express PD‐L1 and PD‐L2 in AA‐affected skin and; (iii) the expressibon of PD‐L1 and PD‐L2 in dermal fibrolasts were regulated by the releasable factors from activated lymphocytes.
      PubDate: 2017-09-04T03:21:08.169369-05:
      DOI: 10.1002/jcp.26134
  • Lipid composition of membrane microdomains isolated detergent‐free from
           PUFA supplemented RAW264.7 macrophages
    • Authors: Christine Hellwing; Feven Tigistu-Sahle, Herbert Fuhrmann, Reijo Käkelä, Julia Schumann
      First page: 2602
      Abstract: Profound alterations in the lipid profile of raft and non‐raft plasma membrane microdomains were found when RAW264.7 macrophages were supplemented with polyunsaturated fatty acids (PUFAs) in physiologically relevant concentrations. For the first time lipids in the detergent‐free isolated membrane domains of phagocytic immune cells were characterized by mass spectrometry. The extent of remodeling of the membrane lipids differed with different n3 and n6 PUFA supplements. The mildest effects were detected for α‐linolenic acid (LNA) and linoleic acid (LA), the C18 precursors of the n3 and n6 families, respectively. When the effects of highly unsaturated PUFAs were compared, eicosapentaenoic acid (EPA) caused more extensive restructuring of membrane lipids than docosahexaenoic acid (DHA) or arachidonic acid (AA). The supplements altered the lipid species composition of both the raft and non‐raft membrane fractions. The rafts containing elevated proportions of highly unsaturated lipid species may relocate sterically incompatible lipids and proteins originally belonging to this microdomain. Such effect was evident for sphingomyelin, which favored non‐rafts instead of rafts after EPA supplementation. The current work suggests that the different functional consequences found previously when supplementing macrophages with either EPA or DHA have their origin in the different effects of these PUFAs on membrane architecture.Profound alterations in the lipid profile of raft and non‐raft plasma membrane microdomains were found when RAW264.7 macrophages were supplemented with polyunsaturated fatty acids (PUFAs) in physiologically relevant concentrations. The supplements altered the lipid species composition of both the raft and non‐raft membrane fractions. Furthermore, the current work suggests that the different functional consequences found previously when supplementing macrophages with either EPA or DHA have their origin in the different effects of these PUFAs on membrane architecture.
      PubDate: 2017-08-30T05:22:20.558434-05:
      DOI: 10.1002/jcp.26138
  • DHQZ‐17, a potent inhibitor of the transcription factor HNF4A,
           suppresses tumorigenicity of head and neck squamous cell carcinoma in vivo
    • Authors: Shilpa Tentu; Kumarswamyreddy Nandarapu, Prakash Muthuraj, Kesavan Venkitasamy, Ganesh Venkatraman, Suresh K. Rayala
      First page: 2613
      Abstract: A series of 2, 3‐dihydroquinazolinone derivatives were synthesized, characterized and their anticancer activity was determined. Among the compounds synthesized and screened, one compound (17) showed potent anticancer activity against human head and neck squamous cell carcinoma cell line, SCC131 and was non‐toxic to normal cells. The compound inhibited the growth of SCC131 cells, with an IC50 of 1.75 μM, triggered apoptotic mode of cell death and caused tumor regression of SCC131 tumor xenografts in athymic mice. To decipher the target for the lead compound, a high throughput qPCR array was performed. Results showed that the compound 17, inhibited the expression of a vital transcription factor HNF4A, involved in regulation of metabolic pathways. Thus, the present work has identified a lead compound 17, with potent anticancer activity, minimal normal cell toxicity and a plausible target and hence definitely holds future prospects as an anticancer agent.A novel small molecule inhibitor for oral cancer.
      PubDate: 2017-09-07T04:05:49.661111-05:
      DOI: 10.1002/jcp.26139
  • Novel role for the testis‐enriched HSPA2 protein in regulating epidermal
           keratinocyte differentiation
    • Authors: Agnieszka Gogler-Pigłowska; Katarzyna Klarzyńska, Damian R. Sojka, Anna Habryka, Magdalena Głowala-Kosińska, Marcin Herok, Mariusz Kryj, Monika Halczok, Zdzisław Krawczyk, Dorota Scieglinska
      First page: 2629
      Abstract: HSPA2, a poorly characterized member of the HSPA (HSP70) chaperone family, is a testis‐enriched protein involved in male germ cell differentiation. Previously, we revealed that HSPA2 is present in human stratified epithelia, including epidermis, however the contribution of this protein to epithelial biology remained unknown. Here, we show for the first time that HSPA2 is expressed in basal epidermal keratinocytes, albeit not in keratinocytes exhibiting features attributed to primitive undifferentiated progenitors, and participates in the keratinocyte differentiation process. We found that HSPA2 is dispensable for protection of HaCaT keratinocytes against heat shock‐induced cytotoxicity. We also shown that lentiviral‐mediated shRNA silencing of HSPA2 expression in HaCaT cells caused a set of phenotypic changes characteristic for keratinocytes committed to terminal differentiation such as reduced clonogenic potential, impaired adhesiveness and increased basal and confluency‐induced expression of differentiation markers. Moreover, the fraction of undifferentiated cells that rapidly adhered to collagen IV was less numerous in HSPA2‐deficient cells than in the control. In a 3D reconstructed human epidermis model, HSPA2 deficiency resulted in accelerated development of a filaggrin‐positive layer. Collectively, our results clearly show a link between HSPA2 expression and maintenance of keratinocytes in an undifferentiated state in the basal layer of the epidermis. It seems that HSPA2 could retain keratinocytes from premature entry into the terminal differentiation process. Overall, HSPA2 appears to be necessary for controlling development of properly stratified epidermis and thus for maintenance of skin homeostasis.In human skin HSPA2 is preferentially expressed in basal epidermal keratinocytes. HSPA2 is dispensable for protection of HaCaT keratinocytes against heat shock‐induced cytotoxicity. HSPA2 participates in the keratinocyte differentiation process.
      PubDate: 2017-09-07T03:56:01.278356-05:
      DOI: 10.1002/jcp.26142
  • Effects of pulsed electromagnetic fields and platelet rich plasma in
           preventing osteoclastogenesis in an in vitro model of osteolysis
    • Authors: Matilde Tschon; Francesca Veronesi, Deyanira Contartese, Maria Sartori, Lucia Martini, Fabrizio Vincenzi, Annalisa Ravani, Katia Varani, Milena Fini
      First page: 2645
      Abstract: Osteolysis is the main limiting cause for the survival of an orthopedic prosthesis and is accompanied by an enhancement in osteoclastogenesis and inflammation, due by wear debris formation. Unfortunately therapeutic treatments, besides revision surgery, are not available. The aim of the present study was to evaluate the effects of Pulsed Electro Magnetic Fields (PEMFs) and platelet rich plasma (PRP), alone or in combination, in an in vitro model of osteolysis. Rats peripheral blood mononuclear cells were cultured on Ultra High Molecular Weight Polyethylene particles and divided into four groups of treatments: (1) PEMF stimulation (12 hr/day, 2.5 mT, 75 Hz, 1.3 ms pulse duration); (2) 10% PRP; (3) combination of PEMFs, and PRP; (4) no treatment. Treatments were performed for 3 days and cell viability, osteoclast number, expression of genes related to osteoclastogenesis and inflammation and production of pro‐inflammatory cytokines were assessed up to 14 days. PEMF stimulation exerted best results because it increased cell viability at early time points and counteracted osteoclastogenesis at 14 days. On the contrary, PRP increased osteoclastogenesis and reduced cell viability in comparison to PEMFs alone. The combination of PEMFs and PRP increased cell viability over time and reduced osteoclastogenesis in comparison to PRP alone. However, these positive results did not exceed the level achieved by PEMF alone. At longer time points PEMF could not counteract osteoclastogenesis increased by PRP. Regarding inflammation, all treatments maintained the production of pro‐inflammatory cytokines at low level, although PRP increased the level of interleukin 1 beta.The aim of the present study was to evaluate the effects of Pulsed ElectroMagnetic Fields (PEMFs) and platelet rich plasma (PRP), alone or in combination, in an in vitro model of osteolysis.
      PubDate: 2017-09-04T03:25:51.779427-05:
      DOI: 10.1002/jcp.26143
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