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Publisher: John Wiley and Sons   (Total: 1616 journals)

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Journal Cover Journal of Cellular Physiology
  [SJR: 1.842]   [H-I: 139]   [6 followers]  Follow
    
   Hybrid Journal Hybrid journal (It can contain Open Access articles)
   ISSN (Print) 0021-9541 - ISSN (Online) 1097-4652
   Published by John Wiley and Sons Homepage  [1616 journals]
  • miR-124 and miR-9 Mediated Downregulation of HDAC5 Promotes Neurite
           Development Through Activating MEF2C- GPM6A Pathway
    • Authors: Xi Gu; Congcong Fu, Lifang Lin, Shuhu Liu, Xiaohong Su, Aili Li, Qiaoqi Wu, Chunhong Jia, Peidong Zhang, Lu Chen, Xinhong Zhu, Xuemin Wang
      Abstract: The class IIa histone deacetylases (HDACs) play important roles in the central nervous system during diverse biological processes such as synaptic plasticity, axon regeneration, cell apoptosis, and neural differentiation. Although it is known that HDAC5 regulates neuronal differentiation, neither the physiological function nor the regulation of HDAC5 in neuronal differentiation is clear. Here, we identify HDAC5 as an inhibitor of neurite elongation and show that HDAC5 is regulated by the brain enriched microRNA miR-124 and miR-9. We discover that HDAC5 inhibits neurite extension both in differentiated P19 cells and primary neurons. We also show that the neuronal membrane glycoprotein GPM6A (M6a) is a direct target gene of HDAC5 regulated transcriptional factor MEF2C. HDAC5 inhibits neurite elongation, acting at least partially via a MEF2C/M6a signaling pathway. We also confirmed the miR-124/miR-9 regulated HDAC5-MEF2C-M6a pathway regulates neurite development in primary neurons. Thus, HDAC5 emerges as a cellular conductor of MEF2C and M6a activity and is regulated by miR-124 and miR-9 to control neurite development. This article is protected by copyright. All rights reserved
      PubDate: 2017-03-23T06:15:41.705303-05:
      DOI: 10.1002/jcp.25927
       
  • Dual-specificity Tyrosine Phosphorylation-regulated Kinase 2 Regulates
           Osteoclast Fusion in a Cell Heterotypic Manner
    • Authors: Gali Guterman-Ram; Milena Pesic, Ayelet Orenbuch, Tal Czeiger, Anastasia Aflalo, Noam Levaot
      Abstract: Monocyte fusion into osteoclasts, bone resorbing cells, plays a key role in bone remodeling and homeostasis; therefore, aberrant cell fusion may be involved in a variety of debilitating bone diseases. Research in the last decade has led to the discovery of genes that regulate osteoclast fusion, but the basic molecular and cellular regulatory mechanisms underlying the fusion process are not completely understood. Here, we reveal a role for Dyrk2 in osteoclast fusion. We demonstrate that Dyrk2 down regulation promotes osteoclast fusion, whereas its overexpression inhibits fusion. Moreover, Dyrk2 also promotes the fusion of foreign-body giant cells, indicating that Dyrk2 plays a more general role in cell fusion. In an earlier study, we showed that fusion is a cell heterotypic process initiated by fusion-founder cells that fuse to fusion-follower cells, the latter of which are unable to initiate fusion. Here, we show that Dyrk2 limits the expansion of multinucleated founder cells through the suppression of the fusion competency of follower cells. This article is protected by copyright. All rights reserved
      PubDate: 2017-03-23T06:15:39.506116-05:
      DOI: 10.1002/jcp.25922
       
  • Involvement of Testicular DAAM1 Expression in Zinc Protection Against
           Cadmium-induced Male Rat Reproductive Toxicity
    • Authors: Marouane Chemek; Massimo Venditti, Sana Boughamoura, Safa Ben Mimouna, Imed Messaoudi, Sergio Minucci
      Abstract: In order to verify the effects of exposure to Cd and Zn on testicular DAAM1 gene and protein expression and also to ascertain their involvement in the protective role of Zn in prevent the testicular toxicity Cd-induced in male offspring rats at adult age after gestational and lactational exposure, male offspring rats, from mothers receiving either tap water, Cd, Zn or Cd + Zn during gestation and lactation periods, were scarified on postnatal days (PND) 70. The reproductive organ (testis, epididymis and vesicle seminal) were collected, weighed and analysed. The results showed that exposure to Cd in utero and through lactation decreased the relative reproductive organ weight, altered the testicular histology at the interstitial and tubular levels, and causing a significant reduction in the daily sperm production (DSP) per testis and per gram of testis, and other then altering the epididymal sperm quality. Furthemore, both mRNA and protein expression of rat testicular DAAM1 were also inhibited in Cd-treated group. Zn supply has completely corrected the most of these toxic effects. Our results imply that Zn could prevent Cd-induced testicular toxicity and sperm quality alteration in adult male rat after gestational and lactational exposure, probably via the restoration of the testicular DAAM1 expression inhibited by Cd. This article is protected by copyright. All rights reserved
      PubDate: 2017-03-23T03:50:58.697216-05:
      DOI: 10.1002/jcp.25923
       
  • Retinoic Acid Receptor-related Orphan Receptor RORα Regulates
           Differentiation and Survival of Keratinocytes During Hypoxia
    • Authors: Hongyu Li; Longjian Zhou, Jun Dai
      Abstract: Low O2 pressures present in the microenvironment of epidermis control keratinocyte differentiation and epidermal barrier function through hypoxia inducible factors (HIFs) dependent gene expression. This study focuses on investigating relations of the retinoic acid receptor-related orphan receptor alpha (RORα) to HIF-1α in keratinocytes under hypoxic conditions. The expression level of RORα is significantly elevated under hypoxia in both human and murine keratinocytes. Gene silencing of RORA attenuates hypoxia-stimulated expression of genes related to late differentiation and epidermal barrier function, and leads to an enhanced apoptotic response. While the hypoxic induction of RORα is dependent on HIF-1α, RORα is in turn critical for nuclear accumulation of HIF-1α and activation of HIF transcriptional activity. These results collectively suggest that RORα functions as an important mediator of HIF-1α activities in regulating keratinocyte differentiation/survival and epidermal barrier function during the oxygen sensing stage. This article is protected by copyright. All rights reserved
      PubDate: 2017-03-23T03:50:27.051248-05:
      DOI: 10.1002/jcp.25924
       
  • Primary Cilium Alterations and Expression Changes of Patched1 Proteins in
           Niemann-pick Type C Disease
    • Authors: Patrizia Formichi; Carla Battisti, Maria Margherita De Santi, Raffaella Guazzo, Sergio Antonio Tripodi, Elena Radi, Benedetta Rossi, Ermelinda Tarquini, Antonio Federico
      Abstract: Niemann-Pick type C disease (NPC) is a disorder characterized by abnormal intracellular accumulation of unesterified cholesterol and glycolipids. Two distinct disease-causing genes have been isolated, NPC1 and NPC2. The NPC1 protein is involved in the sorting and recycling of cholesterol and glycosphingolipids in the late endosomal/lysosomal system. It has extensive homology with the Patched 1 (Ptc1) receptor, a transmembrane protein localized in the primary cilium and involved in the Hedgehog signaling (Shh) pathway.We assessed the presence of NPC1 and Ptc1 proteins and evaluated the relative distribution and morphology of primary cilia in fibroblasts from five NPC1 patients and controls, and in normal fibroblasts treated with 3-ß-[2-(diethylamino)ethoxy]androst-5-en-17-one (U18666A), a cholesterol transport-inhibiting drug that is widely used to mimic NPC. Immunofluorescence and western blot analyses showed a significant decrease in expression of NPC1 and Ptc1 in NPC1 fibroblasts, while they were normally expressed in U18666A-treated fibroblasts. Moreover, fibroblasts from NPC1 patients and U18666A-treated cells showed a lower percentage distribution of primary cilia and a significant reduction in median cilia length with respect to controls. These are the first results demonstrating altered cytoplasmic expression of Ptc1 and reduced number and length of primary cilia, where Ptc1 is located, in fibroblasts from NPC1 patients. We suggest that the alterations in Ptc1 expression in cells from NPC1 patients are closely related to NPC1 expression deficit, while the primary cilia alterations observed in NPC1 and U18666A-treated fibroblasts may represent a secondary event derived from a defective metabolic pathway. This article is protected by copyright. All rights reserved
      PubDate: 2017-03-23T03:46:12.925397-05:
      DOI: 10.1002/jcp.25926
       
  • Mechanism of Prolactin Inhibition of miR-135b via Methylation in Goat
           Mammary Epithelial Cells
    • Authors: Zhi Chen; Jun Luo, ChangHui Zhang, Yue Ma, Shuang Sun, Tianyin Zhang, Juan J. Loor
      Abstract: Prolactin is an important endocrine activator of lactogenesis. This study investigated the function and mechanism of miR-135b in the enhancement of lactation by prolactin in goat mammary epithelial tissue. We utilized S-Poly (T) sequencing to evaluate changes in gene regulation in the goat mammary gland after incubation with 2.5 ug/ml prolactin and 2.5 ug/ml IGF-1 by examining highly expressed miRNAs during early-lactation and late-lactation. The results illustrated that miR-135b is highly expressed in the goat mammary gland during early-lactation and late-lactation, and also after treatment with 2.5 ug/ml prolactin and 2.5 ug/ml IGF-1. We used Q-RT PCR, Western Blot, immunofluorescence, and luciferase reporter assay analysis, and found that PRL was significantly down-regulated in response to the expression of miR-135b in a manner that was functionally related to TAG synthesis via the large tumor suppressor 2 gene (Lats2), an important regulator of adipocyte proliferation via Hippo Signaling. Furthermore, using bisulfite-sequencing PCR (BSP), Q-PCR and Western Blot we discovered an increase in expression of DNMT I (DNA methyl transferase I) in goat mammary epithelial cells with the 2.5 ug/ml PRL incubation, which led to DNA methylation of the CpG island upstream of miR-135b and inhibited the transcription and expression of miR-135b. This article is protected by copyright. All rights reserved
      PubDate: 2017-03-23T03:46:09.289111-05:
      DOI: 10.1002/jcp.25925
       
  • Autophagy Maintains the Integrity of Endothelial Barrier in LPS-induced
           Lung Injury
    • Authors: Dan Zhang; Jian Zhou, Le Chi Ye, Jing Li, Zhenzhou Wu, Yuping Li, Chichi Li
      Abstract: Understanding the role and underlying regulation mechanism of autophagy in lipopolysaccharide-induced lung injury (LPS-LI) may provide potentially new pharmacological targets for treatment of acute lung injury. The aim of this study was to investigate the functional significance of autophagy in LPS-LI. The autophagy of human pulmonary microvascular endothelial cells (HPMVECs) and mice was inhibited before they were challenged with LPS. In vitro, permeability, vitality and the lactate dehydrogenase (LDH) release rate of the cells were detected, the zonula occluden-1 (ZO-1) expression and the stress fiber formation were determined. In vivo, the lung injury was assessed. We found LPS caused high permeability and increased LDH release rate, lowered viability of the cells, inhibited the ZO-1 expression and induced stress fiber formation, these effects were further aggravated by prohibiting the level of autophagy. Consistently, in in vivo experiments, LPS induced serious lung injury, which was reflected as edema, leukocyte infiltration and hemorrhage in lung tissue, and the high concentration of pro-inflammation cytokines tumor necrosis factor (TNF)-α and interleukin (IL)-1β in bronchoalveolar lavage fluid (BALF). Inhibiting autophagy further exacerbated LPS-LI. It appears that autophagy played a protective role in LPS-LI in part through restricting the injury of lung microvascular barrier. This article is protected by copyright. All rights reserved
      PubDate: 2017-03-22T05:01:27.054343-05:
      DOI: 10.1002/jcp.25928
       
  • Gax Suppresses Chemerin/CMKLR1-induced Preadipocyte Biofunctions Through
           the Inhibition of Akt/mTOR and ERK Signaling Pathways
    • Authors: Yunqi Jiang; Ping Liu, Wenlin Jiao, Juan Meng, Jinbo Feng
      Abstract: Adipose tissue is closely associated with angiogenesis and vascular remodeling. Chemerin is involved in inflammatory reaction and vascular dysfunction. However, the mechanisms of chemerin participating in vascular remodeling and whether Growth arrest–specific homeobox (Gax) can effectively intervene it remain obscured. Here, 3T3-F442A preadipocytes were cultured, injected into athymic mice to model fat pads, and treated respectively with Ad-chemerin, Ad-Gax, or specific inhibitors in vitro and in vivo. MTT, flow cytometry, Western blotting, and imunohisto(cyto)-chemistry analyses showed that chemerin enhanced the expression of FABP4 and VEGF, activated Akt/mTOR and ERK pathways, increased the cell percent of S phase, decreased the percent of G0-G1 phase and apoptotic cells, and augmented neovascular density in fat pads. Inversely, Gax suppressed the expression of these adipogenic and vasifactive markers and these signaling proteins, decreased the percent of S phase cells, and increased those of G0-G1 phase and apoptotic cells, and reduced the neovascular density. Our results indicate that chemerin-CMKLR1 activates Akt/mTOR and ERK pathways and facilitates preadipocyte proliferation, adipogenesis, and angiogenesis. Contrarily, Gax weakens the effect of chemerin on preadipocyte biofunctions. This article is protected by copyright. All rights reserved
      PubDate: 2017-03-21T23:01:57.895306-05:
      DOI: 10.1002/jcp.25918
       
  • Mimicking exercise in three-dimensional bioengineered skeletal muscle to
           
    • Authors: Andreas M. Kasper; Daniel C. Turner, Neil R. W. Martin, Adam P. Sharples
      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
       
  • All-trans Retinoic Acid Ameliorates Hepatic Stellate Cell Activation via
           Suppression of Thioredoxin Interacting Protein Expression
    • Authors: Hiroki Shimizu; Toshiaki Tsubota, Keita Kanki, Goshi Shiota
      Abstract: Activation of hepatic stellate cells (HSCs) is the effector factor of hepatic fibrosis and hepatocellular carcinoma (HCC) development. Accumulating evidence suggests that retinoic acids (RAs), derivatives of vitamin A, contribute to prevention of liver fibrosis and carcinogenesis, however, regulatory mechanisms of RAs still remain exclusive. To elucidate RA signaling pathway, we previously performed a genome-wide screening of RA-responsive genes by in silico analysis of RA-response elements, and identified 26 RA-responsive genes. We found that thioredoxin interacting protein (TXNIP), which inhibits antioxidant activity of thioredoxin (TRX), was downregulated by all-trans retinoic acid (ATRA). In the present study, we demonstrate that ATRA ameliorates activation of HSCs through TXNIP suppression. HSC activation was attenuated by TXNIP downregulation, whereas potentiated by TXNIP upregulation, indicating that TXNIP plays a crucial role in activation of HSCs. Notably, we showed that TXNIP-mediated HSC activation was suppressed by antioxidant N-acetylcysteine. In addition, ATRA treatment or downregulation of TXNIP clearly declined oxidative stress levels in activated HSCs. These data suggest that ATRA plays a key role in inhibition of HSC activation via suppressing TXNIP expression, which reduces oxidative stress levels. This article is protected by copyright. All rights reserved
      PubDate: 2017-03-21T07:27:39.140761-05:
      DOI: 10.1002/jcp.25921
       
  • TGF-β Inhibits Osteogenesis by Upregulating the Expression of Ubiquitin
           Ligase SMURF1 via MAPK-ERK Signaling
    • Authors: Xuewu Sun; Ziang Xie, Yan Ma, Xin Pan, Jiying Wang, Zhijun Chen, Peihua Shi
      Abstract: High incidence of osteoporotic fractures emphasizes the necessity of developing effective measures to promote osteogenesis. In our study, we investigated a possible role of MAPK-ERK signaling in the TGF-β-mediated osteoblastic differentiation. Our results indicated that TGF-β activated the MAPK-ERK pathway and inhibited osteogenesis in mesenchymal pluripotent cell line, C3H10T1/2, and preosteoblastic cell line, MC3T3 cells. And the downregulation of MAPK-ERK signaling using pharmacological inhibitor U0126 and RNA interference rescued osteoblast differentiation suppressed by TGF-β, which was confirmed by Alkaline phosphatase (ALP) staining and alizarrn red staining and the enhanced expression of osteogenesic markers. Western blotting analysis indicated that TGF-β induced protein expression of E3 ubiquitin-protein ligase SMURF1, which contributed to the degradation of RUNX2 and SMAD1 as evidenced by SMURF1 inhibition using RNA interference and proteasome inhibitor MG132. Moreover, we observed that the expression of SMURF1 was decreased, while that of SMAD1 and RUNX2 increased by MAPK-ERK inhibitor U0126 in TGF-β-treated differentiating preosteoblasts, suggesting that MAPK-ERK regulated the transcription of osteogenesis-related genes. Furthermore, a synergistic effect between U0126 and bone morphogenic protein (BMP)-2 on osteoblast differentiation and bone formation was observed both in cell cultures and experimental animals. In conclusion, our results revealed that TGF-β inhibited osteoblastic differentiation by inducing the MAPK-ERK pathway which upregulated the expression of ubiquitin ligase SMURF1 and resulted in reduced presence of osteogenic proteins. In addition, the potentiation of BMP-2 on osteogenic activity by ERK1/2 inhibitor U0126 suggests that it may have potential clinical utility for promoting osteogenesis in bone fracture repair. This article is protected by copyright. All rights reserved
      PubDate: 2017-03-21T07:27:31.075954-05:
      DOI: 10.1002/jcp.25920
       
  • Regulation Role of CRTC3 in Skeletal Muscle and Adipose Tissue
    • Authors: Jiaqi Liu; Ziye Xu, Weiche Wu, Yizhen Wang, Tizhong Shan
      Abstract: The cyclic adenosine monophosphate (cAMP) - protein kinase A (PKA) signaling pathway plays important role in regulating energy homeostasis. Many of the effects of the cAMP-PKA signaling is mediated through the cAMP responsive element binding protein (CREB) and its coactivator CREB-regulated transcription coactivators (CRTCs). CRTC3 is a member of CRTCs family proteins and plays important roles in glucose and energy metabolism. Previous studies shown that global knockout of CRTC3 enhances oxygen consumption and energy expenditure and subsequently protects the knockout animal against obesity. In skeletal muscle, CRTC3 affects lipid and glycogen metabolism and mitochondrial biogenesis. In white adipocytes, CRTC3 regulates GLUT4 expression and glucose uptake. More recently, the localization and function of CRTC3 in brown fat have been reported. In this review, we mainly discuss the regulatory role of CRTC3 in skeletal muscle and adipose tissues. This article is protected by copyright. All rights reserved
      PubDate: 2017-03-21T07:21:11.234825-05:
      DOI: 10.1002/jcp.25917
       
  • Transplantation of Mesenchymal Stem Cells Overexpressing IL10 Attenuates
           Cardiac Impairments in Rats with Myocardial Infarction
    • Authors: Xin Meng; Jianping Li, Ming Yu, Jian Yang, Minjuan Zheng, Jinzhou Zhang, Chao Sun, Hongliang Liang, Liwen Liu
      Abstract: Mesenchymal stem cell (MSC) has been well known to exert therapeutic potential for patients with myocardial infarction (MI). In addition, interleukin-10 (IL10) could attenuate MI through suppressing inflammation. Thus, the combination of MSC implantation with IL10 delivery may extend health benefits to ameliorate cardiac injury after MI. Here we established overexpression of IL10 in bone marrow-derived MSC through adenoviral transduction. Cell viability, apoptosis and IL10 secretion under ischemic challenge in vitro were examined. In addition, MSC was transplanted into the injured hearts in a rat model of MI. Four weeks after the MI induction, myocardial infarction, cardiac functions, apoptotic cells and inflammation cytokines were assessed. In response to in vitro oxygen-glucose deprivation (OGD), IL10 overexpression in MSC (Ad.IL10-MSC) enhanced cell viability, decreased apoptosis and increased IL10 secretion. Consistently, the implantation of Ad.IL10-MSCs into MI animals resulted in more reductions in myocardial infarct size, cardiac impairment and cell apoptosis, compared to the individual treatments of either MSC or IL10 administration. Moreover, the attenuation of both systemic and local inflammations was most prominent for Ad.IL10-MSC treatment. IL10 overexpression and MSC may exert a synergistic anti-inflammatory effect to alleviate cardiac injury after MI. This article is protected by copyright. All rights reserved
      PubDate: 2017-03-21T07:20:58.420555-05:
      DOI: 10.1002/jcp.25919
       
  • Identification and Characterization of Site-specific N-glycosylation in
           the Potassium Channel Kv3.1b
    • Authors: Paul Christian Vicente; Jin Young Kim, Jeong-Ju Ha, Min-Young Song, Hyun-Kyung Lee, Dong-Hyun Kim, Jin-Sung Choi, Kang-Sik Park
      Abstract: The potassium ion channel Kv3.1b is a member of a family of voltage-gated ion channels that are glycosylated in their mature form. In the present study, we demonstrate the impact of N-glycosylation at specific asparagine residues on the trafficking of the Kv3.1b protein. Large quantities of asparagine 229 (N229)-glycosylated Kv3.1b reached the plasma membrane, whereas N220-glycosylated and unglycosylated Kv3.1b were mainly retained in the endoplasmic reticulum (ER). These ER-retained Kv3.1b proteins were susceptible to degradation when co-expressed with calnexin, whereas Kv3.1b pools located at the plasma membrane were resistant. Mass spectrometry analysis revealed a complex type Hex3HexNAc4Fuc1 glycan as the major glycan component of the N229-glycosylated Kv3.1b protein, as opposed to a high-mannose type Man8GlcNAc2 glycan for N220-glycosylated Kv3.1b. Taken together, these results suggest that trafficking-dependent roles of the Kv3.1b potassium channel are dependent on N229 site-specific glycosylation and N-glycan structure, and operate through a mechanism whereby specific N-glycan structures regulate cell surface expression. This article is protected by copyright. All rights reserved
      PubDate: 2017-03-21T07:20:43.960399-05:
      DOI: 10.1002/jcp.25915
       
  • HDAC1 and HDAC3 Underlie Dynamic H3K9 Acetylation During Embryonic
           Neurogenesis and in Schizophrenia-like Animals
    • Authors: Josef Večeřa; Eva Bártová, Jana Krejčí, Soňa Legartová, Denisa Komůrková, Jana Rudá-Kučerová, Tibor Štark, Eva Dražanová, Tomáš Kašpárek, Alexandra Šulcová, Frank J. Dekker, Wiktor Szymanski, Christian Seiser, Georg Weitzer, Raphael Mechoulam, Vincenzo Micale, Stanislav Kozubek
      Abstract: Although histone acetylation is one of the most widely studied epigenetic modifications, there is still a lack of information regarding how the acetylome is regulated during brain development and pathophysiological processes. We demonstrate that the embryonic brain (E15) is characterized by an increase in H3K9 acetylation as well as decreases in the levels of HDAC1 and HDAC3. Moreover, experimental induction of H3K9 hyperacetylation led to the overexpression of NCAM in the embryonic cortex and depletion of Sox2 in the subventricular ependyma, which mimicked the differentiation processes. Inducing differentiation in HDAC1-deficient mouse ESCs resulted in early H3K9 deacetylation, Sox2 downregulation and enhanced astrogliogenesis, whereas neuro-differentiation was almost suppressed. Neuro-differentiation of (wt) ESCs was characterized by H3K9 hyperacetylation that was associated with HDAC1 and HDAC3 depletion. Conversely, the hippocampi of schizophrenia-like animals showed H3K9 deacetylation that was regulated by an increase in both HDAC1 and HDAC3. The hippocampi of schizophrenia-like brains that were treated with the cannabinoid receptor-1 inverse antagonist AM251 expressed H3K9ac at the level observed in normal brains. Together, the results indicate that co-regulation of H3K9ac by HDAC1 and HDAC3 is important to both embryonic brain development and neuro-differentiation as well as the pathophysiology of a schizophrenia-like phenotype. This article is protected by copyright. All rights reserved
      PubDate: 2017-03-16T07:06:05.26055-05:0
      DOI: 10.1002/jcp.25914
       
  • Rapamycin Attenuates BAFF-extended Proliferation and Survival via
           Disruption of mTORC1/2 Signaling in Normal and Neoplastic B-lymphoid Cells
           
    • Authors: Qingyu Zeng; Shanshan Qin, Hai Zhang, Beibei Liu, Jiamin Qin, Xiaoxue Wang, Ruijie Zhang, Chunxiao Liu, Xiaoqing Dong, Shuangquan Zhang, Shile Huang, Long Chen
      Abstract: B cell activating factor from the TNF family (BAFF) stimulates B-cell proliferation and survival, but excessive BAFF promotes the development of aggressive B cells leading to malignant and autoimmune diseases. Recently, we have reported that rapamycin, a macrocyclic lactone, attenuates human soluble BAFF (hsBAFF)-stimulated B-cell proliferation/survival by suppressing mTOR-mediated PP2A-Erk1/2 signaling pathway. Here, we show that the inhibitory effect of rapamycin on hsBAFF-promoted B cell proliferation/survival is also related to blocking hsBAFF-stimulated phosphorylation of Akt, S6K1 and 4E-BP1, as well as expression of survivin in normal and B-lymphoid (Raji and Daudi) cells. It appeared that both mTORC1 and mTORC2 were involved in the inhibitory activity of rapamycin, as silencing raptor or rictor enhanced rapamycin's suppression of hsBAFF-induced survivin expression and proliferation/viability in B cells. Also, PP242, an mTORC1/2 kinase inhibitor, repressed survivin expression and cell proliferation/viability more potently than rapamycin (mTORC1 inhibitor) in B cells in response to hsBAFF. Of interest, ectopic expression of constitutively active Akt (myr-Akt) or constitutively active S6K1 (S6K1-ca), or downregulation of 4E-BP1 conferred resistance to rapamycin's attenuation of hsBAFF-induced survivin expression and B-cell proliferation/viability, whereas overexpression of dominant negative Akt (dn-Akt) or constitutively hypophosphorylated 4E-BP1 (4EBP1-5A), or downregulation of S6K1, or co-treatment with Akt inhibitor potentiated the inhibitory effects of rapamycin. The findings indicate that rapamycin attenuates excessive hsBAFF-induced cell proliferation/survival via blocking mTORC1/2 signaling in normal and neoplastic B-lymphoid cells. Our data underscore that rapamycin may be a potential agent for preventing excessive BAFF-evoked aggressive B-cell malignancies and autoimmune diseases. This article is protected by copyright. All rights reserved
      PubDate: 2017-03-16T07:00:45.056107-05:
      DOI: 10.1002/jcp.25913
       
  • Knockout of ATG5 Leads to Malignant Cell Transformation and Resistance to
           Src Family Kinase Inhibitor PP2
    • Authors: Sung-Hee Hwang; Byeal-I Han, Michael Lee
      Abstract: Autophagy can either promote or inhibit cell death in different cellular contexts. In this study, we investigated the role of autophagy in ATG5 knockout (KO) cell line established using CRISPR/Cas9 system. In ATG5 KO cells, RT-PCR and immunoblot of LC3 confirmed the functional gene knockout. We found that knockout of ATG5 significantly increased proliferation of NIH 3T3 cells. In particular, autophagy deficiency enhanced susceptibility to cellular transformation as determined by an in vitro clonogenic survival assay and a soft agar colony formation assay. We also found that ATG5 KO cells had a greater migration ability as compared to wild-type (WT) cells. Moreover, ATG5 KO cells were more resistant to treatment with a Src family tyrosine kinase inhibitor (PP2) than WT cells were. Cyto-ID Green autophagy assay revealed that PP2 failed to induce autophagy in ATG5 KO cells. PP2 treatment decreased the percentage of cells in the S and G2/M phases among WT cells but had no effect on cell cycle distribution of ATG5 KO cells, which showed a high percentage of cells in the S and G2/M phases. Additionally, the proportion of apoptotic cells significantly decreased after treatment of ATG5 KO cells with PP2 in comparison with WT cells. We found that expression levels of p53 were much higher in ATG5 KO cells. The ATG5 KO seems to lead to compensatory upregulation of the p53 protein because of a decreased apoptosis rate. Taken together, our results suggest that autophagy deficiency can lead to malignant cell transformation and resistance to PP2. This article is protected by copyright. All rights reserved
      PubDate: 2017-03-15T06:55:54.137758-05:
      DOI: 10.1002/jcp.25912
       
  • 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 Kumar Onteru
      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 h 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 1ng/mL FSH and 10 ng/mL IGF-1 for 144 h. 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. This article is protected by copyright. All rights reserved
      PubDate: 2017-03-15T06:55:50.136304-05:
      DOI: 10.1002/jcp.25909
       
  • Autophagy Mediates Cytotoxicity of Human Colorectal Cancer Cells Treated
           with Garcinielliptone FC
    • Authors: Shen-Jeu Won; Cheng-Hsin Yen, Ting-Yu Lin, Ya-Fen Jiang-Shieh, Chun-Nan Lin, Jyun-Ti Chen, Chun-Li Su
      Abstract: The tautomeric pair of garcinielliptone FC (GFC) is a novel tautomeric pair of polyprenyl benzophenonoid isolated from the pericarps of Garcinia subelliptica Merr. (G. subelliptica, Clusiaceae), a tree with abundant sources of polyphenols. Our previous report demonstrated that GFC induced apoptosis on various types of human cancer cell lines including chemoresistant human colorectal cancer HT-29 cells. In the present study, we observed that many autophagy-related genes in GFC-treated HT-29 cells were up- and down-regulated using a cDNA microarray containing oncogenes and kinase genes. GFC-induced autophagy of HT-29 cells was confirmed by observing the formation of acidic vesicular organelles, LC3 puncta, and double-membrane autophagic vesicles using flow cytometry, confocal microscopy, and transmission electron microscopy, respectively. Inhibition of AKT/mTOR/P70S6K signaling as well as formation of Atg5-Atg12 and PI3K/Beclin-1 complexes were observed using Western blot. Administration of autophagy inhibitor (3-methyladenine and shRNA Atg5) and apoptosis inhibitor Z-VAD showed that the GFC-induced autophagy was cytotoxic form and GFC-induced apoptosis enhanced GFC-induced autophagy. Our data suggest the involvement of autophagy and apoptosis in GFC-induced anticancer mechanisms of human colorectal cancer. This article is protected by copyright. All rights reserved
      PubDate: 2017-03-15T06:55:39.862725-05:
      DOI: 10.1002/jcp.25910
       
  • Probiotics Are a Good Choice in Remission of Inflammatory Bowel Diseases:
           A Meta Analysis and Systematic Review
    • Authors: Mahboube Ganji-Arjenaki; Mahmoud Rafieian-Kopaei
      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< 0.01). Analysis of 18 trials revealed that probiotics in patients with Ulcerative colitis (UC) in different conditions have significant effects (p = 0.007). VSL#3 probiotics in patients with UC had significant effect (p 
      PubDate: 2017-03-15T06:55:34.436935-05:
      DOI: 10.1002/jcp.25911
       
  • Artesunate Inhibits RANKL-induced Osteoclastogenesis and Bone Resorption
           In Vitro and Prevents LPS-induced Bone Loss In Vivo
    • Authors: Cheng-Ming Wei; Qian Liu, Fang-Ming Song, Xi-Xi Lin, Yi-Ji Su, Jiake Xu, Lin Huang, Shao-Hui Zong, Jin-Min Zhao
      Abstract: Osteoclasts are multinuclear giant cells responsible for bone resorption in lytic bone diseases such as osteoporosis, arthritis, periodontitis, and bone tumors. Due to the severe side-effects caused by the currently available drugs, a continuous search for novel bone-protective therapies is essential. Artesunate (Art), the water-soluble derivative of artemisinin has been investigated owing to its anti-malarial properties. However, its effects in osteoclastogenesis have not yet been reported. In this study, Art was shown to inhibit the nuclear factor-κB ligand (RANKL)-induced osteoclastogenesis, the mRNA expression of osteoclastic-specific genes, and resorption pit formation in a dose-dependent manner in primary bone marrow-derived macrophages cells (BMMs). Furthermore, Art markedly blocked the RANKL-induced osteoclastogenesis by attenuating the degradation of IκB and phosphorylation of NF-κB p65. Consistent with the in vitro results, Art inhibited lipopolysaccharide (LPS)-induced bone resorption by suppressing the osteoclastogenesis. Together our data demonstrated that Art inhibits RANKL-induced osteoclastogenesis by suppressing the NF-κB signaling pathway and that it is a promising agent for the treatment of osteolytic diseases. This article is protected by copyright. All rights reserved
      PubDate: 2017-03-15T06:50:30.84516-05:0
      DOI: 10.1002/jcp.25907
       
  • Pannexin Hemichannels: A Novel Promising Therapy Target for Oxidative
           Stress Related Diseases
    • Authors: Jin Xu; Linxi Chen, Lanfang Li
      Abstract: Pannexins, which contain three subtypes: pannexin-1, pannexin-2 and pannexin-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. This article is protected by copyright. All rights reserved
      PubDate: 2017-03-14T05:36:14.897847-05:
      DOI: 10.1002/jcp.25906
       
  • Lipopolysaccharide Can Modify Differentiation and Immunomodulatory
           Potential of Periodontal Ligament Stem Cells via ERK1,2 Signaling
    • Authors: Tamara Kukolj; Drenka Trivanović, Ivana Okić Djordjević, Slavko Mojsilović, Jelena Krstić, Hristina Obradović, Srdja Janković, Juan Francisco Santibanez, Aleksandra Jauković, Diana Bugarski
      Abstract: Lipopolysaccharide (LPS) is a pertinent deleterious factor in oral microenvironment for cells which are carriers of regenerative processes. The aim of this study was to investigate the emerging in vitro effects of LPS (E.coli) on human periodontal ligament stem cell (PDLSC) functions and associated signaling pathways. We demonstrated that LPS did not affect immunophenotype, proliferation, viability and cell cycle of PDLSCs. However, LPS modified lineage commitment of PDLSCs inhibiting osteogenesis by downregulating Runx2, ALP and Ocn mRNA expression, while stimulating chondrogenesis and adipogenesis by upregulating Sox9 and PPARγ mRNA expression. LPS promoted myofibroblast-like phenotype of PDLSCs, since it significantly enhanced PDLSCs contractility, as well as protein and/or gene expression of TGF-β, fibronectin, α-SMA and NG2. LPS also increased protein and gene expression levels of anti-inflammatory COX-2 and pro-inflammatory IL-6 molecules in PDLSCs. Inhibition of peripheral blood mononuclear cells (MNCs) transendothelial migration in presence of LPS-treated PDLSCs was accompanied by the reduction of CD29 expression within MNCs. However, LPS treatment did not change the inhibitory effect of PDLSCs on mitogen-stimulated proliferation of CD4+ and the ratio of CD4+CD25high/CD4+CD25low lymphocytes. LPS-treated PDLSCs did not change the frequency of CD34+ and CD45+ cells, but decreased the frequency of CD33+ and CD14+ myeloid cells within MNCs. Moreover, LPS treatment attenuated the stimulatory effect of PDLSCs on CFC activity of MNCs, predominantly the CFU-GM number. The results indicated that LPS-activated ERK1,2 was at least partly involved in the observed effects on PDLSC differentiation capacity, acquisition of myofibroblastic attributes and changes of their immunomodulatory features. This article is protected by copyright. All rights reserved
      PubDate: 2017-03-14T05:36:09.988576-05:
      DOI: 10.1002/jcp.25904
       
  • Mitochondrial ROS-induced ERK1/2 Activation and HSF2-mediated AT1R
           Upregulation Are Required for Doxorubicin-induced Cardiotoxicity
    • Authors: Chih-Yang Huang; Jia-Yi Chen, Chia-Hua Kuo, Pei-Ying Pai, Tsung-Jung Ho, Tung-Sheng Chen, Fu-Jen Tsai, V. Vijaya Padma, Wei-Wen Kuo, Chih-Yang Huang
      Abstract: Doxorubicin (DOX), one useful chemotherapeutic agent, is limited in clinical use because of its serious cardiotoxicity. Growing evidence suggests that angiotensin receptor blockers (ARBs) have cardioprotective effects in DOX-induced cardiomyopathy. However, the detailed mechanisms underlying the action of ARBs on the prevention of DOX-induced cardiomyocyte cell death have yet to be investigated. Our results showed that angiotensin II receptor type I (AT1R) plays a critical role in DOX-induced cardiomyocyte apoptosis. We found that MAPK signaling pathways, especially ERK1/2, participated in modulating AT1R gene expression through DOX-induced mitochondrial ROS release. These results showed that several potential heat shock binding elements (HSE), which can be recognized by heat shock factors (HSFs), located at the AT1R promoter region. HSF2 markedly translocated from the cytoplasm to the nucleus when cardiomyocytes were damaged by DOX. Furthermore, the DNA binding activity of HSF2 was enhanced by DOX via deSUMOylation. Overexpression of HSF2 enhanced DOX-induced cardiomyocyte cell death as well. Taken together, we found that DOX induced mitochondrial ROS release to activate ERK-mediated HSF2 nuclear translocation and AT1R upregulation causing DOX-damaged heart failure in vitro and in vivo. This article is protected by copyright. All rights reserved
      PubDate: 2017-03-14T05:36:05.410059-05:
      DOI: 10.1002/jcp.25905
       
  • Visualization of Stimulus-specific Heterogeneous Activation of Individual
           Vascular Smooth Muscle Cells in Aortic Tissues
    • Authors: Satoshi Komatsu; Toshio Kitazawa, Mitsuo Ikebe
      Abstract: Intercellular communication among autonomic nerves, endothelial cells (ECs) and vascular smooth muscle cells (VSMCs) plays a central role in an uninterrupted regulation of blood flow through vascular contractile machinery. Impairment of this communication is linked to development of vascular diseases such as hypertension, cerebral/coronary vasospasms, aortic aneurism, and erectile dysfunction. Although the basic concept of the communication as a whole has been studied, the spatiotemporal correlation of ECs/VSMCs in tissues at the cellular level is unknown. Here we show a unique VSMC response to ECs during contraction and relaxation of isolated aorta tissues through visualization of spatiotemporal activation patterns of smooth muscle myosin II. ECs in the intimal layer dictate the stimulus-specific heterogeneous activation pattern of myosin II in VSMCs within distinct medial layers. Myosin light chain (MLC) phosphorylation (active form of myosin II) gradually increases towards outer layers (approximately 3 fold higher MLC phosphorylation at the outermost layer than that of the innermost layer), presumably by release of an intercellular messenger, nitric oxide (NO). Our study also demonstrates that the MLC phosphorylation at the outermost layer in spontaneously hypertensive rats (SHR) during NO-induced relaxation is quite high and approximately 10 fold higher than that of its counterpart, the Wister-Kyoto rats (WKY), suggesting that the distinct pattern of myosin II activation within tissues is important for vascular protection against elevated blood pressure. This article is protected by copyright. All rights reserved
      PubDate: 2017-03-11T06:35:31.524848-05:
      DOI: 10.1002/jcp.25903
       
  • Fibroblast Dynamics as an In Vitro Screening Platform for Anti-fibrotic
           Drugs in Primary Myelofibrosis
    • Authors: Ciprian Tomuleasa; Sonia Selicean, Grigore Gafencu, Bobe Petrushev, Laura Pop, Cristian Berce, Anca Jurj, Adrian Trifa, Ana-Maria Rosu, Sergiu Pasca, Lorand Magdo, Mihnea Zdrenghea, Delia Dima, Alina Tanase, Ioana Frinc, Anca Bojan, Ioana Berindan- Neagoe, Gabriel Ghiaur, Stefan O. Ciurea
      Abstract: Although the cause for bone marrow fibrosis in patients with myelofibrosis remains controversial, it has been hypothesized that it is caused by extensive fibroblast proliferation under the influence of cytokines generated by the malignant megakaryocytes. Moreover, there is no known drug therapy which could reverse the process. We studied the fibroblasts in a novel system using the hanging drop method, evaluated whether the fibroblasts obtain from patients are part of the malignant clone of not and, using this system, we screen a large library of FDA-approved drugs to identify potential drugs candidates that might be useful in the treatment of this disease, specifically which would inhibit fibroblast proliferation and the development of bone marrow fibrosis. We have found that the BM fibroblasts are not part of the malignant clone, as previously suspected and two immunosuppressive medications – cyclosporine and mycophenolate mophetil, as most potent suppressors of the fibroblast collagen production thus potentially inhibitors of bone marrow fibrosis production in myelofibrosis. This article is protected by copyright. All rights reserved
      PubDate: 2017-03-11T06:35:27.369179-05:
      DOI: 10.1002/jcp.25902
       
  • 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
      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. This article is protected by copyright. All rights reserved
      PubDate: 2017-03-10T10:51:59.901222-05:
      DOI: 10.1002/jcp.25901
       
  • Osteoactivin Regulates Head and Neck Squamous Cell Carcinoma Invasion by
           Modulating Matrix Metalloproteases
    • Authors: Oneida A. Arosarena; Eric W. Barr, Ryan Thorpe, Hilary Yankey, Joseph T. Tarr, Fayez F. Safadi
      Abstract: Nearly 60% of patients with head and neck squamous cell carcinoma (HNSCC) die of metastases or locoregional recurrence. Metastasis is mediated by cancer cell migration and invasion, which are in part dependent on extracellular matrix degradation by matrix metalloproteinases. Osteoactivin (OA) overexpression plays a role in metastases in several malignancies, and has been shown to upregulate matrix metalloproteinase (MMP) expression and activity.ObjectivesTo determine how OA modulates MMP expression and activity in HNSCC, and to investigate OA effects on cell invasion.Materials and MethodsWe assessed effects of OA treatment on MMP mRNA and protein expression, as well as gelatinase and caseinolytic activity in HNSCC cell lines. We assessed the effects of OA gene silencing on MMP expression, gelatinase and caseinolytic activity, and cell invasion.ResultsOA treatment had differential effects on MMP mRNA expression. OA treatment upregulated MMP-10 expression in UMSCC14a (p = 0.0431) and SCC15 (p 
      PubDate: 2017-03-10T10:51:56.547133-05:
      DOI: 10.1002/jcp.25900
       
  • HMGB1 down-regulation mediates terameprocol vascular anti-proliferative
           effect in experimental pulmonary hypertension
    • Authors: Rita Nogueira-Ferreira; Manuel J. Ferreira-Pinto, Ana Filipa Silva, Rui Vitorino, Joana Justino, Raquel Costa, Daniel Moreira-Gonçalves, Jean-François Quignard, Thomas Ducret, Jean-Pierre Savineau, Adelino F. Leite-Moreira, Rita Ferreira, Tiago Henriques-Coelho
      Abstract: Pulmonary arterial hypertension (PAH) is a progressive disease with a poor prognosis. Pulmonary artery smooth muscle cells (PASMCs) play a crucial role in PAH pathophysiology, displaying a hyperproliferative, and apoptotic-resistant phenotype. In the present study, we evaluated the potential therapeutic role of terameprocol (TMP), an inhibitor of cellular proliferation and promoter of apoptosis, in a well-established pre-clinical model of PAH induced by monocrotaline (MCT) and studied the biological pathways modulated by TMP in PASMCs. Wistar rats injected with MCT or saline (SHAM group) were treated with TMP or vehicle. On day 21 after injection, we assessed bi-ventricular hemodynamics and cardiac and pulmonary morphometry. The effects of TMP on PASMCs were studied in a primary culture isolated from SHAM and MCT-treated rats, using an iTRAQ-based proteomic approach to investigate the molecular pathways modulated by this drug. In vivo, TMP significantly reduced pulmonary and cardiac remodeling and improved cardiac function in PAH. In vitro, TMP inhibited proliferation and induced apoptosis of PASMCs. A total of 65 proteins were differentially expressed in PASMCs from MCT rats treated with TMP, some of which involved in the modulation of transforming growth factor beta pathway and DNA transcription. Anti-proliferative effect of TMP seems to be explained, at least in part, by the down-regulation of the transcription factor HMGB1. Our findings support the beneficial role of TMP in PAH and suggest that it may be an effective therapeutic option to be considered in the clinical management of PAH.TMP-reduced pulmonary and cardiac remodeling and hemodynamic features of PAH. Its vascular anti-proliferative effect seems to be mediated by the protein HMGB1. TMP may be an effective therapeutic option in the clinical management of PAH.
      PubDate: 2017-03-09T16:55:32.77611-05:0
      DOI: 10.1002/jcp.25763
       
  • Phosphoregulation of K+-Cl− Cotransporters During Cell Swelling:
           Novel Insights
    • Authors: Rachelle Frenette-Cotton; Andrée-Anne Marcoux, Alexandre P. Garneau, Micheline Noel, Paul Isenring
      Abstract: The K+-Cl− cotransporters (KCCs) belong to the cation-Cl− cotransporter family and consist of 4 isoforms and many splice variants. Their main role is to promote electroneutral efflux of K+ and Cl− ions across the surface of many cell types and, thereby, to regulate intracellular ion concentration, cell volume and epithelial salt movement. These transport systems are induced by an increase in cell volume and are less active at lower intracellular [Cl−] (Cli), but the mechanisms at play are still ill-defined. In this work, we have exploited the Xenopus laevis expression system to study the role of lysine-deficient protein kinases (WNKs), protein phosphatases 1 (PP1s) and SPS1-related proline/alanine-rich kinase (SPAK) in KCC4 regulation during cell swelling. We have found that WNK4 and PP1 regulate KCC4 activity as part of a common signalling module, but that they do not exert their effects through SPAK or carrier dephosphorylation. We have also found that the phosphatases at play include PP1α and PP1γ1, but that WNK4 acts directly on the PP1s instead of the opposite. Unexpectedly, however, both cell swelling and a T926A substitution in the C-terminus of full-length KCC4 led to higher levels of heterologous K+-Cl− cotransport and overall carrier phosphorylation. These results imply that the response to cell swelling must also involve allosteric-sensitive kinase-dependent phosphoacceptor sites in KCC4. They are thus partially inconsistent with previous models of KCC regulation. This article is protected by copyright. All rights reserved
      PubDate: 2017-03-09T06:55:26.222564-05:
      DOI: 10.1002/jcp.25899
       
  • Comparative Proteomic Profiling of Human Osteoblast-Derived Extracellular
           Matrices Identifies Proteins Involved in Mesenchymal Stromal Cell
           Osteogenic Differentiation and Mineralization
    • Authors: Marta Baroncelli; Bram C. Eerden, Yik-Yang Kan, Rodrigo D. Alves, Jeroen A. Demmers, Jeroen Peppel, Johannes P. Leeuwen
      Abstract: The extracellular matrix (ECM) is a dynamic component of tissue architecture that physically supports cells and actively influences their behavior. In the context of bone regeneration, cell-secreted ECMs have become of interest as they reproduce tissue-architecture and modulate the promising properties of mesenchymal stem cells (MSCs). We have previously created an in vitro model of human osteoblast-derived devitalized ECM that was osteopromotive for MSCs. The aim of this study was to identify ECM regulatory proteins able to modulate MSC differentiation to broaden the spectrum of MSC clinical applications. To this end, we created 2 additional models of devitalized ECMs with different mineralization phenotypes. Our results showed that the ECM derived from osteoblast-differentiated MSCs had increased osteogenic potential compared to ECM derived from undifferentiated MSCs and non-ECM cultures. Proteomic analysis revealed that structural ECM proteins and ribosomal proteins were upregulated in the ECM from undifferentiated MSCs. A similar response profile was obtained by treating osteoblast-differentiating MSCs with Activin-A. Extracellular proteins were upregulated in Activin-A ECM, whereas mitochondrial and membrane proteins were downregulated. In summary, this study illustrates that the composition of different MSC-secreted ECMs is important to regulate the osteogenic differentiation of MSCs. These models of devitalized ECMs could be used to modulate MSC properties to regulate bone quality. This article is protected by copyright. All rights reserved
      PubDate: 2017-03-08T06:25:39.157919-05:
      DOI: 10.1002/jcp.25898
       
  • Cytokine networks and their association with Helicobacter pylori infection
           in gastric carcinoma
    • Authors: Vahid Bagheri; Bahram Memar, Amir Abbas Momtazi, Amirhossein Sahebkar, Mehran Gholamin, Mohammad Reza Abbaszadegan
      Abstract: Cytokine networks as dynamic networks are pivotal aspects of tumor immunology, especially in gastric cancer (GC), in which infection, inflammation, and antitumor immunity are key elements of disease progression. In this review, we describe functional roles of well-known GC-modulatory cytokines, highlight the functions of cytokines with more recently described roles in GC, and emphasize the therapeutic potential of targeting the complex cytokine milieu. We also focus on the role of Helicobacter pylori (HP)-induced inflammation in GC and discuss how HP-induced chronic inflammation can lead to the induction of stem cell hyperplasia, morphological changes in gastric mucosa and GC development.In this review, we describe functional roles of well-known gastric cancer (GC)-modulatory cytokines, highlight the functions of cytokines with more recently described roles in GC, and emphasize the therapeutic potential of targeting the complex cytokine milieu. We also focus on the role of Helicobacter pylori (HP)-induced inflammation in GC and discuss how HP-induced chronic inflammation can lead to GC.
      PubDate: 2017-03-07T08:02:50.346867-05:
      DOI: 10.1002/jcp.25822
       
  • Role of Wnt/β-catenin Signaling Regulatory microRNAs in the
           Pathogenesis of Colorectal Cancer
    • Authors: Farzad Rahmani; Amir Avan, Seyed Isaac Hashemy, Seyed Mahdi Hassanian
      Abstract: Colorectal cancer (CRC) is one of the leading causes of cancer death worldwide. In more than 90% of all CRC patients, the master oncogenic Ras-Wnt signaling axis is over-activated. MicroRNAs (miRNAs) are potential novel diagnostic and prognostic biomarkers as well as therapeutic targets for several cancers including lung, breast, gastric and colorectal cancers. Oncogenic or tumor suppressor miRNAs modulate tumor cells proliferation, cell cycle progression, angiogenesis, invasion and metastasis through regulating oncogenic pathways including Wnt/β-catenin signaling. This review summarizes the current knowledge about the role of Wnt/β-catenin signaling regulatory miRNAs in the pathogenesis of colorectal cancer for a better understanding and hence a better management of this disease. This article is protected by copyright. All rights reserved
      PubDate: 2017-03-07T06:41:42.944516-05:
      DOI: 10.1002/jcp.25897
       
  • Bone Loss in C57BL/6J-OlaHsd Mice, a Substrain of C57BL/6J Carrying
           Mutated Alpha-Synuclein and Multimerin-1 Genes
    • Authors: Tamar Liron; Bitya Raphael, Sahar Hiram-Bab, Itai A Bab, Yankel Gabet
      Abstract: The inbred mouse strain C57BL/6 is commonly used for the generation of transgenic mouse and is a well-established strain in bone research. Different vendors supply different substrains of C57BL/6J as wild-type animals when genetic drift did not incur any noticeable phenotype. However, we sporadically observed drastic differences in the bone phenotype of ‘WT’ C57BL/6J mice originating from different labs and speculated that these variations are attributable, at least in part, to the variation between C57BL/6J substrains, which is often overlooked. C57BL/6J-OlaHsd is a commonly used substrain that despite a well-defined deletion in the alpha-synuclein (Snca) and multimerin-1 (Mmrn1) genes, was reported to display no obvious phenotype and is used as WT control. Here we compared the bone phenotype of C57BL/6J-OlaHsd (6J-OLA) to C57BL/6J-RccHsd (6J-RCC) and to the original C57BL/6J (6J-JAX). Using µCT analysis, we found that 6J-OLA mice display a significantly lower trabecular bone mass compared to 6J-RCC and 6J-JAX. PCR analysis revealed that both the Snca and Mmrn1 genes are expressed in bone tissue of 6J-RCC animals but not of 6J-OLA mutants, suggesting either or both genes play a role in bone metabolism. In vitro analysis demonstrated increase in osteoclasts number and decreased osteoblast mineralization in cells derived from 6J-OLA compared with 6J-RCC. Our data may shed light on unexplained differences in basal bone measurements between different research centers and reiterate the importance of specifying the exact substrain type. In addition, our findings describe the physiological role for Mmrn1 and/or Snca in bone remodeling. This article is protected by copyright. All rights reserved
      PubDate: 2017-03-07T06:41:40.875338-05:
      DOI: 10.1002/jcp.25895
       
  • The Prognostic Value of MGMT Promoter Methylation in Glioblastoma: A
           Meta-analysis of Clinical Trials
    • Authors: Maryam Moradi; Afsane Bahrami, Soodabeh ShahidSales, Marjan Joodi, Mona Joudi Mashhad, Seyed Mahdi Hassanian, Kazem Anvari, Amir Avan
      Abstract: The DNA repair protein O6-Methylguanine-DNA methyltransferase (MGMT) is suggested to be associated with resistance to alkylating agents such as Temozolomide which is being used in treatment of patients with glioblastoma (GBM). Therefore we evaluated the associations between MGMT promoter methylation and prognosis of patients with glioblastoma (GBM). Data were extracted from publications in Embase, PubMed, and the Cochrane Library. Data on overall survival (OS), progression-free survival (PFS), and MGMT methylation status were obtained and 4097 subjects were enrolled. Data from 34 studies showed that MGMT methylated patients had better OS, compared to GBM unmethylated patients (pooled HRs, 0.494; 95% CI 0.412–0.591; P = 0.001). Meta-analysis of 10 eligible studies reporting on PFS, demonstrated that MGMT promoter methylation was not significantly associated with better PFS (pooled HRs, 0.653; 95% CI 0.414–1.030; P = 0.067). GBM patients with MGMT methylation were associated with longer overall survival, although this effect was not detected for PFS. Moreover we performed further analysis in patients underwent a comprehensive imaging evaluation. This data showed a significant association with better OS and PFS, although further studies are warranted to assess the value of emerging marker in prospective setting in patients with glioblastoma as a risk stratification biomarker in clinical management of the patients. This article is protected by copyright. All rights reserved
      PubDate: 2017-03-07T06:41:12.443631-05:
      DOI: 10.1002/jcp.25896
       
  • Gingival Spheroids Possess Multilineage Differentiation Potential
    • Authors: Rajasekaran Subbarayan; Dinesh Murugan Girija, Suresh Ranga Rao
      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 article is protected by copyright. All rights reserved
      PubDate: 2017-03-07T04:30:31.198486-05:
      DOI: 10.1002/jcp.25894
       
  • Reciprocal regulation between βTrCP and Smurf1 suppresses proliferative
           capacity of liver cancer cells
    • Authors: Yue Zhang; Wenhua Wang, Si Cai, Yan Chen, Qinwan Wang, Qiuhui Pan, Fenyong Sun, Jiayi Wang
      Abstract: We previously reported that both the ubiquitin E3 ligases βTrCP (beta-transducin repeat-containing E3 ubiquitin protein ligase) and Smurf1 (SMAD-specific E3 ubiquitin protein ligase 1) play similar antitumorigenic roles in liver cancer cells. However, whether and how they are reciprocally regulated remains elusive. Here, we show that βTrCP interacts with Smurf1 through the 7 × tryptophan (W) aspartic acid (D)(WD) 40 and the region homologous to the E6-AP carboxyl terminus (HECT) domains, which are the E3 ligase domains of βTrCP and Smurf1, respectively. The E3 ligase domains of βTrCP and Smurf1 are also critical for maintaining the protein expressions of Smurf1 and βTrCP. Moreover, a positive correlation between βTrCP and Smurf1 was also revealed by tissue microarray analysis, indicating that this relationship might be important in liver cancer. Further, we found that Smurf1 increases the protein stability of βTrCP, possibly by reducing autoubiquitination of βTrCP, and vice versa. Interestingly, such effects depended on the presence of E3 ligase domains. Importantly, depletion of Smurf1- or βTrCP-enhanced proliferative capacity of liver cancer cells could be partially reversed by overexpression of wild-type βTrCP or Smurf1 but not their E3 ligase-dead mutants. Collectively, a reciprocal post-translational regulation between βTrCP and Smurf1 has been uncovered in this study. Simultaneous enhancement of βTrCP and Smurf1 functions might be helpful in the treatment of liver cancer.In the present study, we identified a close interaction between βTrCP and Smurf1, uncovering a reciprocal post-translational regulation between RING and HECT E3s in liver cancer cells. Simultaneous stimulation of βTrCP and Smurf1 functions may help to conquer liver cancer.
      PubDate: 2017-03-06T11:00:27.779784-05:
      DOI: 10.1002/jcp.25780
       
  • Cancer Stem Cells (CSCs) in melanoma: There's smoke, but is there
           fire'
    • Authors: Constance E. Brinckerhoff
      Abstract: Cancer stem cells (CSCs), also called Tumor Initiating Cells (TICs), can be defined as cancer cells that are present within solid tumors or hematological cancers, which have characteristics associated with normal stem cells, but which can give rise to all cell types found in a particular cancer sample. CSCs, therefore, are transformed stem cells, which can self-renew, differentiate into diverse progenies, and drive continuous tumor growth (Kreso & Dick, , Cell Stem Cell, 14:275-291; Schatton et al., , Nature, 451:345-349; Villani, Sabbatino, Ferrone, & Ferrone, , Melanoma Management, 2:109-114; Zhou et al., , Drug Discovery, 8:806–823) (Fig. ).The cancer stem cell hypothesis. Tumor cells are heterogeneous and only the tumor-initiating cells have the ability to proliferate extensively, give rise to differentiated cells and form new tumors. There are various therapeutic strategies that could target tumor-initiating cells. Killing these cells could be achieved by inhibiting their survival pathways. Some of the survival pathways that are used by tumor-initiating cells could also be used by the bulk of the tumor, so agents targeting these pathways are expected to kill more than just tumor-initiating cells (adapted from Zhou et al. 2009)
      PubDate: 2017-03-06T10:55:23.80228-05:0
      DOI: 10.1002/jcp.25796
       
  • Swelling-induced Chloride Current in Glioblastoma Proliferation, Migration
           and Invasion
    • Authors: Raymond Wong; Wenliang Chen, Xiao Zhong, James T. Rutka, Zhong-Ping Feng, Hong-Shuo Sun
      Abstract: Glioblastoma (GBM) remains as the most common and aggressive brain tumor. The survival of GBM has been linked to the aberrant activation of swelling-induced chloride current ICl,swell. In this study, we investigated the effects of ICl,swell on cell viability, proliferation and migration in the human GBM cell lines, U251 and U87, using a combination of patch clamp electrophysiology, MTT, colony formation, wound healing assays and Western immunoblotting. First, we showed that the specific inhibitor of ICl,swell, DCPIB, potently reduced the ICl,swell in U87 cells. Next, in both U87 and U251 cells, we found that DCPIB reduced GBM viability, proliferation, colony formation, migration and invasion. In addition, our Western immunoblot assay showed that DCPIB-treated U251 cells had a reduction in JAK2, STAT3, and Akt phosphorylation, thus suggesting that DCPIB potentially suppresses GBM functions through inhibition of the JAK2/STAT3 and PI3K/Akt signaling pathways. Therefore, the ICl,swell may be a potential drug target for GBM. This article is protected by copyright. All rights reserved
      PubDate: 2017-03-06T07:16:18.060666-05:
      DOI: 10.1002/jcp.25891
       
  • 2-Methoxyestradiol Impacts on Amino Acids-mediated Metabolic Reprogramming
           in Osteosarcoma Cells by Interaction with NMDA Receptor
    • Authors: Magdalena Gorska-Ponikowska; Ugo Perricone, Alicja Kuban-Jankowska, Giosue Lo Bosco, Giampaolo Barone
      Abstract: Deregulation of serine and glycine metabolism, have been identified to function as metabolic regulators in supporting tumor cell growth. The role of serine and glycine in regulation of cancer cell proliferation is complicated, dependent on concentrations of amino acids and tissue-specific. D-serine and glycine are coagonists of N-methyl-D-aspartate receptor subunit GRIN1. Importantly, NMDA receptors are widely expressed in cancer cells and play an important role in regulation of cell death, proliferation and metabolism of numerous malignancies.The aim of the present work was to associate the metabolism of glycine and D-serine with the anticancer activity of 2-methoxyestradiol. 2-methoxyestradiol is a potent anticancer agent but also a physiological 17β- estradiol metabolite.In the study we have chosen two malignant cell lines expressing functional GRIN1 receptors, i.e. osteosarcoma 143B and breast cancer MCF7. We used MTS assay, migration assay, flow cytometric analyses, western blotting and immunoprecipitation techniques as well as molecular modeling studies.We have demonstrated the extensive crosstalk between the deregulated metabolic network and cancer cell signaling. Herein, we observed an anticancer effect of high concentrations of glycine and D-serine in osteosarcoma cells. In contrast, the amino acids when used at low, physiological concentrations induced the proliferation and migration of osteosarcoma and breast cancer cells. Importantly, the pro-cancergogenic effects of both glycine and D-serine where abrogated by the usage of 2-methoxyestradiol at both physiological and pharmacological relevant concentrations. The obtained data confirmed that 2-methoxyestradiol may be a physiological anticancer molecule. This article is protected by copyright. All rights reserved
      PubDate: 2017-03-06T07:11:38.253422-05:
      DOI: 10.1002/jcp.25888
       
  • Effects of Reduced Natural Background Radiation on Drosophila melanogaster
           Growth and Development as Revealed by the FLYINGLOW Program
    • Authors: P. Morciano; R. Iorio, D. Iovino, F. Cipressa, G. Esposito, A. Porrazzo, L. Satta, E. Alesse, M.A. Tabocchini, G. Cenci
      Abstract: Natural background radiation of Earth and cosmic rays played a relevant role during the evolution of living organisms. However, how chronic low doses of radiation can affect biological processes is still unclear. Previous data have indicated that cells grown at the Gran Sasso Underground Laboratory (LNGS, L'Aquila) of National Institute of Nuclear Physics (INFN) of Italy, where the dose rate of cosmic rays and neutrons is significantly reduced with respect to the external environment, elicited an impaired response against endogenous damage as compared to cells grown outside LNGS. This suggests that environmental radiation contributes to the development of defense mechanisms at cellular level. To further understand how environmental radiation affects metabolism of living organisms, we have recently launched the FLYINGLOW program that aims at exploiting Drosophila melanogaster as a model for evaluating the effects of low doses/dose rates of radiation at the organismal level. Here, we will present a comparative data set on lifespan, motility and fertility from different Drosophila strains grown in parallel at LNGS and in a reference laboratory at the University of L'Aquila. Our data suggest the reduced radiation environment can influence Drosophila development and, depending on the genetic background, may affect viability for several generations even when flies are moved back to normal background radiation. As flies are considered a valuable model for human biology, our results might shed some light on understanding the effect of low dose radiation also in humans. This article is protected by copyright. All rights reserved
      PubDate: 2017-03-06T07:10:51.877725-05:
      DOI: 10.1002/jcp.25889
       
  • Targeting the 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
      Abstract: The V-Ki-ras2 Kirsten rat sarcoma viral oncogene homolog (KRAS) is an important pathways that 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 most 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. This article is protected by copyright. All rights reserved
      PubDate: 2017-03-06T07:10:27.219352-05:
      DOI: 10.1002/jcp.25890
       
  • Exhaustive Training Leads to Hepatic Fat Accumulation
    • Authors: Alisson L. da Rocha; Ana P. Pinto, Giovana R. Teixeira, Bruno C. Pereira, Luciana C. Oliveira, Adriana C. Silva, Gustavo P. Morais, Dennys E. Cintra, José R. Pauli, Adelino S. R. da Silva
      Abstract: Recently, we demonstrated that an overtraining (OT) protocol for mice based on downhill running sessions increased the hepatic phosphorylation of 70-kDa ribosomal protein S6 kinase 1 (S6K1; Thr389), a downstream target of the mammalian target of rapamycin complex 1 (mTORC1). In liver, the overactivation of the Akt/mTORC1 pathway induces lipogenesis via regulation of the action of sterol regulatory element binding protein-1 (SREBP-1) at multiple steps. Herein, we verified the effects of three running OT models with same external load (i.e., the product between intensity and volume of training), but performed in downhill, uphill and without inclination, on the proteins related to the mTORC1 signaling pathway, the protein content of the SREBP-1, ACC, and FAS, and the morphological characteristics of C57BL/6 mouse livers. In summary, the downhill running-induced OT model up-regulated the levels of major proteins of the mTORC1 signaling pathway, the protein levels of SREBP-1 (p125 precursor) and induced signs of cell swelling accompanied by acute inflammation. The other two OT protocols performed uphill and without inclination did not modulate the most analyzed molecular proteins, but induced hepatic morphological alterations, suggesting an acute pathological adaptation. The three OT models induced hepatic fat accumulation. J. Cell. Physiol. 9999: 1–10, 2016. © 2016 Wiley Periodicals, Inc.The downhill running-induced overtraining model increased the levels of major proteins of the mTOR signaling pathway in mouse livers despite high levels of pAMPK (Thr172). The three overtraining models increased the percentage of fat in liver.
      PubDate: 2017-03-03T12:20:50.318332-05:
      DOI: 10.1002/jcp.25625
       
  • Association between AXL, Hippo Transducers, and Survival Outcomes in Male
           Breast Cancer
    • Authors: Anna Di Benedetto; Marcella Mottolese, Francesca Sperati, Cristiana Ercolani, Luigi Di Lauro, Laura Pizzuti, Patrizia Vici, Irene Terrenato, Abeer M. Shaaban, Matthew P. Humphries, Sreekumar Sundara-Rajan, Maddalena Barba, Valerie Speirs, Ruggero De Maria, Marcello Maugeri-Saccà
      Abstract: Male breast cancer (MBC) is an uncommon malignancy. We have previously reported that the expression of the Hippo transducers TAZ/YAP and their target CTGF was associated with inferior survival in MBC patients. Preclinical evidence demonstrated that Axl is a transcriptional target of TAZ/YAP. Thus, we herein assessed AXL expression to further investigate the significance of active TAZ/YAP-driven transcription in MBC. For this study, 255 MBC samples represented in tissue microarrays were screened for AXL expression, and 116 patients were included. The association between categorical variables was verified by the Pearson's Chi-squared test of independence (2-tailed) or the Fisher Exact test. The relationship between continuous variables was tested with the Pearson's correlation coefficient. The Kaplan–Meier method was used for estimating survival curves, which were compared by log-rank test. Factors potentially impacting 10-year and overall survival were verified in Cox proportional regression models. AXL was positively associated with the TAZ/CTGF and YAP/CTGF phenotypes (P = 0.001 and P = 0.002, respectively). Patients with TAZ/CTGF/AXL- or YAP/CTGF/AXL-expressing tumors had inferior survival compared with non-triple-positive patients (log rank P = 0.042 and P = 0.048, respectively). The variables TAZ/CTGF/AXL and YAP/CTGF/AXL were adverse factors for 10-year survival in the multivariate Cox models (HR 2.31, 95%CI:1.02–5.22, P = 0.045, and HR 2.27, 95%CI:1.00–5.13, P = 0.050). Nearly comparable results were obtained from multivariate analyses of overall survival. The expression pattern of AXL corroborates the idea of the detrimental role of TAZ/YAP activation in MBC. Overall, Hippo-linked biomarkers deserve increased attention in this rare disease. J. Cell. Physiol. 9999: 1–7, 2017. © 2016 Wiley Periodicals, Inc.(1) The expression pattern of AXL corroborates the idea of TAZ/YAP activation in male breast cancer; (2) patients whose tumors had the TAZ/CTGF/AXL and YAP/CTGF/AXL molecular backgrounds experienced adverse survival outcomes compared with their negative counterparts.
      PubDate: 2017-03-03T12:20:48.256102-05:
      DOI: 10.1002/jcp.25745
       
  • TUDCA: An Agonist of the Bile Acid Receptor GPBAR1/TGR5 With
           Anti-Inflammatory Effects in Microglial Cells
    • Authors: Natalia Yanguas-Casás; M. Asunción Barreda-Manso, Manuel Nieto-Sampedro, Lorenzo Romero-Ramírez
      Abstract: Bile acids are steroid acids found in the bile of mammals. The bile acid conjugate tauroursodeoxycholic acid (TUDCA) is neuroprotective in different animal models of stroke and neurological diseases. We have previously shown that TUDCA has anti-inflammatory effects on glial cell cultures and in a mouse model of acute neuroinflammation. We show now that microglial cells (central nervous system resident macrophages) express the G protein-coupled bile acid receptor 1/Takeda G protein-coupled receptor 5 (GPBAR1/TGR5) in vivo and in vitro. TUDCA binding to GPBAR1/TGR5 caused an increase in intracellular cAMP levels in microglia that induced anti-inflammatory markers, while reducing pro-inflammatory ones. This anti-inflammatory effect of TUDCA was inhibited by small interference RNA for GPBAR1/TGR5 receptor, as well as by treatment with a protein kinase A (PKA) inhibitor. In the mouse model of acute neuroinflammation, treating the animals with TUDCA was clearly anti-inflammatory. TUDCA biased the microglial phenotype in vivo and in vitro toward the anti-inflammatory. The bile acid receptor GPBAR1/TGR5 could be a new therapeutic target for pathologies coursing with neuroinflammation and microglia activation, such as traumatic brain injuries, stroke, or neurodegenerative diseases. TUDCA and other GPBAR1/TGR5 agonists need to be further investigated, to determine their potential in attenuating the neuropathologies associated with microglia activation. J. Cell. Physiol. 9999: 1–15, 2016. © 2016 Wiley Periodicals, Inc.We provide evidence that tauroursodeoxycholic acid (TUDCA) exerts anti-inflammatory effects through its binding to GPBAR1/TGR5, and the consequent increase in intracellular cAMP levels in microglia. Besides, TUDCA treatment biased microglia towards the anti-inflammatory phenotype under pro-inflammatory conditions in vitro and in vivo. TUDCA and other GPBAR1/TGR5 agonists are potential therapeutic targets for the treatment of neuropathologies associated with microglia activation.
      PubDate: 2017-03-03T12:20:37.338218-05:
      DOI: 10.1002/jcp.25742
       
  • Systemic and Local Adipose Tissue in Knee Osteoarthritis
    • Authors: Elisa Belluzzi; Hamza El Hadi, Marnie Granzotto, Marco Rossato, Roberta Ramonda, Veronica Macchi, Raffaele De Caro, Roberto Vettor, Marta Favero
      Abstract: Osteoarthritis is a common chronic joint disorder affecting older people. The knee is the major joint affected. The symptoms of osteoarthritis include limited range of motion, joint swelling, and pain causing disability. There are no disease modifying drugs available, and treatments are mainly focused on pain management. Total knee replacement performed at the end stage of the disease is considered the only cure available. It has been found that obese people have an increased risk to develop not only knee but also hand osteoarthritis. This supports the concept that adipose tissue might be related to osteoarthritis not only through overloading. As matter of fact, obesity induces a low grade systemic inflammatory state characterized by the production and secretion of several adipocytokines that may have a role in osteoarthritis development. Furthermore, hypertension, impaired glucose, and lipid metabolism, which are comorbidities associated with obesity, have been shown to alter the joint tissue homeostasis. Moreover, infrapatellar fat pad in the knee has been demonstrated to be a local source of adipocytokines and potentially contribute to osteoarthritis pathogenesis. Here, we discuss the role of systemic and local adipose tissue in knee osteoarthritis. J. Cell. Physiol. 9999: 1–8, 2016. © 2016 Wiley Periodicals, Inc.Obesity is a risk factor for osteoarthritis development related to systemic inflammation other than joint overloading. Infrapatellar fat pad seems to have a role in the pathogenesis of osteoarthritis producing adipocytokines. Future research should be aimed to understand a possible crosstalk between local and systemic adipose tissue.
      PubDate: 2017-03-03T12:20:32.39605-05:0
      DOI: 10.1002/jcp.25716
       
  • Tumor Necrosis Factor-α Increases Claudin-1, 4, and 7 Expression in
           Tubular Cells: Role in Permeability Changes
    • Authors: Yasaman Amoozadeh; Qinghong Dan, Shaista Anwer, Hsiao Han Huang, Vanessa Barbieri, Faiza Waheed, Mazharul Maishan, Katalin Szászi
      Abstract: Tumor necrosis factor-α (TNFα), is a pathogenic cytokine in kidney disease that alters expression of claudins in tubular cells. Previously we showed that in LLC-PK1 cells TNFα caused a biphasic change in transepithelial resistance (TER) consisting of an early drop and recovery, followed by a late increase. However, the underlying mechanisms and the role of specific claudins in the TER effect remained incompletely understood. Here we sought to define how TNFα affects claudins 1, 4, and 7 in tubular cells and to correlate their changes with the TER effect. We show that TNFα elevates total and surface levels of Cldn-1, 4, and 7, and increases their mRNA expression through the ERK and JNK pathways. Further, JNK is also important for TNFα-induced changes in claudin-2 expression. Continuous monitoring of TER using Electric cell-substrate impedance sensing (ECIS) reveals that the two phases of the TNFα effect are differently regulated. Specifically, inhibition of the ERK or JNK pathways prevent the late TER increase, but not the early TER effect. Silencing experiments also show that Cldn-1 is necessary for the early TNFα-induced TER change, while all three claudins appear to contribute to the late TER increase. In summary, we define a central role for ERK and JNK in TNFα-induced altered claudin expression and barrier tightening. Together, our current and previous works show that the TNFα-induced early TER effect requires claudin-1, while claudin-2 decrease is a significant mediator of the late TER increase, and elevation in claudin-1, 4, and 7 contribute to a smaller extent. J. Cell. Physiol. 9999: 1–11, 2016. © 2016 Wiley Periodicals, Inc.We demonstrate that TNFα elevates levels of claudins 1,4, and 7 and define a central role for ERK and JNK in this effect. Using ECIS we correlate these signalling pathways and claudins with the biphasic TER changes induced by TNFα and show their differential roles in the early and late phases.
      PubDate: 2017-03-03T12:20:27.160254-05:
      DOI: 10.1002/jcp.25736
       
  • Polydeoxyribonucleotides (PDRNs) From Skin to Musculoskeletal Tissue
           Regeneration via Adenosine A2A Receptor Involvement
    • Authors: Francesca Veronesi; Dante Dallari, Giacomo Sabbioni, Chiara Carubbi, Lucia Martini, Milena Fini
      Abstract: Polydeoxyribonucleotides (PDRNs) are low molecular weight DNA molecules of natural origin that stimulate cell migration and growth, extracellular matrix (ECM) protein production, and reduce inflammation. Most preclinical and clinical studies on tissue regeneration with PDRNs focused on skin, and only few are about musculoskeletal tissues. Starting from an overview on skin regeneration studies, through the analysis of in vitro, in vivo, and clinical studies (1990–2016), the present review aimed at defining the effects of PDRN and their mechanisms of action in the regeneration of musculoskeletal tissues. This would also help future researches in this area. A total of 29 studies were found by PubMed and www.webofknowledge.com searches: 20 were on skin (six in vitro, six in vivo, one vitro/vivo, seven clinical studies), while the other nine regarded bone (one in vitro, two in vivo, one clinical studies), cartilage (one in vitro, one vitro/vivo, two clinical studies), or tendon (one clinical study) tissues regeneration. PDRNs improved cell growth, tissue repair, ECM proteins, physical activity, and reduced pain and inflammation, through the activation of adenosine A2A receptor. PDRNs are currently used for bone, cartilage, and tendon diseases, with a great variability regarding the PDRN dosage to be used in clinical practice, while the dosage for skin regeneration is well established. PDRNs are usually administered from a minimum of three to a maximum of five times and they act trough the activation of A2A receptor. Further studies are advisable to confirm the effectiveness of PDRNs and to standardize the PDRN dose. J. Cell. Physiol. 9999: 1–9, 2016. © 2016 Wiley Periodicals, Inc.Polydeoxyribonucleotides (PDRNs) are low molecular weight DNA molecules of natural origin that stimulate cell migration and growth, extracellular matrix (ECM) protein production, and reduce inflammation. Most preclinical and clinical studies on tissue regeneration with PDRNs focused on skin, and only few are about musculoskeletal tissues. A total of 29 studies were found by PubMed and www.webofknowledge.com searches: 20 were on skin (six in vitro, six in vivo, one vitro/vivo, seven clinical studies), while the other nine regarded bone (one in vitro, two in vivo, one clinical studies), cartilage (one in vitro, one vitro/vivo, two clinical studies), or tendon (one clinical study) tissues regeneration. PDRNs improved cell growth, tissue repair, ECM proteins, physical activity, and reduced pain and inflammation, through the activation of adenosine A2A receptor.
      PubDate: 2017-03-03T10:50:26.615404-05:
      DOI: 10.1002/jcp.25663
       
  • Nuclear Phosphoinositide Regulation of Chromatin
    • Authors: Bree L. Hamann; Raymond D. Blind
      Abstract: Phospholipid signaling has clear connections to a wide array of cellular processes, particularly in gene expression and in controlling the chromatin biology of cells. However, most of the work elucidating how phospholipid signaling pathways contribute to cellular physiology have studied cytoplasmic membranes, while relatively little attention has been paid to the role of phospholipid signaling in the nucleus. Recent work from several labs has shown that nuclear phospholipid signaling can have important roles that are specific to this cellular compartment. This review focuses on the nuclear phospholipid functions and the activities of phospholipid signaling enzymes that regulate metazoan chromatin and gene expression. In particular, we highlight the roles that nuclear phosphoinositides play in several nuclear-driven physiological processes, such as differentiation, proliferation, and gene expression. Taken together, the recent discovery of several specifically nuclear phospholipid functions could have dramatic impact on our understanding of the fundamental mechanisms that enable tight control of cellular physiology. This article is protected by copyright. All rights reserved
      PubDate: 2017-03-03T08:00:32.895593-05:
      DOI: 10.1002/jcp.25886
       
  • A Biophysical View on von Willebrand Factor Activation
    • Authors: Achim Löf; Jochen P. Müller, Maria A. Brehm
      Abstract: The process of hemostatic plug formation at sites of vascular injury crucially relies on the large multimeric plasma glycoprotein von Willebrand factor (VWF) and its ability to recruit platelets to the damaged vessel wall via interaction of its A1 domain with platelet GPIbα. Under normal blood flow conditions, VWF multimers exhibit a very low binding affinity for platelets. Only when subjected to increased hydrodynamic forces, which primarily occur in connection with vascular injury, VWF can efficiently bind to platelets. This force-regulation of VWF's hemostatic activity is not only highly intriguing from a biophysical perspective, but also of eminent physiological importance. On the one hand, it prevents undesired activity of VWF in intact vessels that could lead to thromboembolic complications and on the other hand, it enables efficient VWF-mediated platelet aggregation exactly where needed.Here, we review recent studies that mainly employed biophysical approaches in order to elucidate the molecular mechanisms underlying the complex mechano-regulation of the VWF-GPIbα interaction. Their results led to two main hypotheses: first, intramolecular shielding of the A1 domain is lifted upon force-induced elongation of VWF; second, force-induced conformational changes of A1 convert it from a low-affinity to a high-affinity state. We critically discuss these hypotheses and aim at bridging the gap between the large-scale behavior of VWF as a linear polymer in hydrodynamic flow and the detailed properties of the A1-GPIbα bond at the single-molecule level. This article is protected by copyright. All rights reserved
      PubDate: 2017-03-03T07:45:36.45689-05:0
      DOI: 10.1002/jcp.25887
       
  • Analysis of Extracellular Vesicles Produced in the Biofilm by the
           Dimorphic Yeast Pichia fermentans
    • Authors: Federica Leone; Lorenza Bellani, Simonetta Mucciflora, Lucia Giorgetti, Paolo Bongioanni, Marcella Simili, BE Maserti, Renata Del Carratore
      Abstract: The yeast Pichia fermentans DISAABA 726 strain (P. fermentans) is a dimorphic yeast that under different environmental conditions may switch from a yeast-like to pseudohyphal morphology. We hypothesize that exosomes-like vesicles (EV) could mediate this rapid modification. As EV are membrane-derived vesicles carrying lipids, proteins, mRNAs and microRNAs and have been recognized as important mediators of intercellular communication. Although it has been assumed for a long time that fungi release EV, knowledge of their functions is still limited. In this work we analyze P. fermentans EV production, during growth in two different media containing urea (YCU) or methionine (YCM) where yeast-like or pseudohyphal morphology, are produced. We developed a procedure to extract EV from the neighboring biofilm, which is faster and more efficient as compared to the widely used ultracentrifugation method. Differences in morphology and RNA content of EV, suggest that they might have an active role during dimorphic transition as response to the growth conditions. Our findings are coherent with a general state of hypoxic stress of the pseudohyphal cells. This article is protected by copyright. All rights reserved
      PubDate: 2017-03-03T07:45:26.290121-05:
      DOI: 10.1002/jcp.25885
       
  • Myocardial Proteases and Cardiac Remodeling
    • Authors: S Riaz; A Zeidan, F Mraiche
      Abstract: Cardiac hypertrophy (CH), characterized by the enlargement of cardiomyocytes, fibrosis and apoptosis, is one of the leading causes of death worldwide. Despite the advances in cardiovascular research, there remains a need to further investigate the signaling pathways that mediate CH in order to identify novel therapeutic targets. One of the hallmarks of CH is the remodeling of the extracellular matrix (ECM). Multiple studies have shown an important role of cysteine proteases and matrix metalloproteinases (MMPs) in the remodeled heart. This review focuses on the role of cysteine cathepins and MMPs in cardiac remodeling. This article is protected by copyright. All rights reserved
      PubDate: 2017-03-03T01:26:27.798376-05:
      DOI: 10.1002/jcp.25884
       
  • Transcription Factor EGR1 Promotes Differentiation of Bovine Skeletal
           Muscle Satellite Cells by Regulating MyoG Gene Expression
    • Authors: Weiwei Zhang; Huili Tong, Ziheng Zhang, Shuli Shao, Dan Liu, Shufeng Li, Yunqin Yan
      Abstract: The transcription factor, early growth response 1 (EGR1), has important roles in various cell types in response to different stimuli. EGR1 is thought to be involved in differentiation of bovine skeletal muscle-derived satellite cells (MDSCs); however, the precise effects of EGR1 on differentiation of MDSCs and its mechanism of action remain unknown. In the present study, a time course of EGR1 expression and the effects of EGR1 on MDSC differentiation were determined. The results demonstrated that the expression of EGR1 mRNA and protein increased significantly in differentiating MDSCs relative to that in proliferating cells. Over-expression of the EGR1 gene in MDSCs promoted their differentiation and inhibited proliferation. Conversely, knock-down of EGR1 inhibited differentiation of MDSCs and promoted their proliferation, indicating that EGR1 promotes MDSC differentiation. Moreover, over-expression of EGR1 in MDSCs increased the expression of MyoG mRNA and protein, whereas its knock-down had the opposite effect. Furthermore, ChIP-PCR analyses demonstrated that EGR1 could bind directly to its putative binding site within the promoter region of MyoG, and determination of ERG1 subcellular localization in MDSCs demonstrated that it could relocate to the nucleus, indicating MyoG is likely an EGR1 target gene whose expression is positively regulated by this transcription factor. In conclusion, EGR1 can promote MDSC differentiation through positive regulation of MyoG gene expression. This article is protected by copyright. All rights reserved
      PubDate: 2017-03-03T01:26:26.56571-05:0
      DOI: 10.1002/jcp.25883
       
  • Deregulated ALG-2/HEBP2 axis alters microtubule dynamics and mitotic
           spindle behavior to stimulate cancer development
    • Authors: Juan Qin; Yang Yang, Siqi Gao, Yang Liu, Fan Yu, Yunqiang Zhou, Rui Lyu, Min Liu, Xinqi Liu, Dengwen Li, Jun Zhou
      Abstract: Cancer cells are characterized by genomic instability, resulting in the accumulation of mutations that promote cancer progression. One way that genomic instability can arise is through improper regulation of the microtubule cytoskeleton that impacts the function of the mitotic spindle. In this study, we have identified a critical role for the interaction between apoptosis-linked gene 2 (ALG-2) and heme-binding protein 2 (HEBP2) in the above processes. Our data show that the gene copy numbers and mRNA levels for both ALG-2 and HEBP2 are significantly upregulated in breast and lung cancer. Coexpression of ALG-2 and HEBP2 markedly increases the cytoplasmic pool of ALG-2 and alters the subcellular distribution of HEBP2. Our data further reveal that abnormality in the ALG-2/HEBP2 interaction impairs spindle orientation and positioning during mitosis. In addition, this complex appears to modulate the dynamic properties of microtubules in cancer cells. These finding thus uncover an important function for deregulated ALG-2/HEBP2 axis in cancer development by influencing microtubule dynamics and spindle behavior, providing novel insight into the etiology and pathogenesis of cancer.In this study, we uncover a previously unrecognized role for deregulated ALG-2/HEBP2 axis in cancer development. Mechanistically, this is mediated by altered microtubule dynamics and impaired spindle orientation and positioning.
      PubDate: 2017-03-01T10:50:25.470059-05:
      DOI: 10.1002/jcp.25754
       
  • Proinflammatory signaling functions of thrombin in cancer
    • Authors: Safieh Ebrahimi; Farzad Rahmani, Reihane Behnam-Rassouli, Fatemeh Hoseinkhani, Mohammad Reza Parizadeh, Mohammad Reza Keramati, Majid Khazaie, Amir Avan, Seyed Mahdi Hassanian
      Abstract: Thrombin-induced activation of protease-activated receptors (PARs) represents a link between inflammation and cancer. Proinflammatory signaling functions of thrombin are associated with several inflammatory diseases including neurodegenerative, cardiovascular, and of special interest in this review cancer. Thrombin-induced inflammatory responses up-regulates expression of cytokines, adhesion molecules, angiogenic factors, and matrix-degrading proteases that facilitate tumor cells proliferation, angiogenesis, invasion, and metastasis. This review summarizes the current knowledge about the mechanisms of thrombin-mediated proinflammatory responses in cancer pathology for a better understanding and hence a better management of this disease.Thrombin-induced inflammatory responses up-regulates expression of cytokines, adhesion molecules, angiogenic factors, and matrix-degrading proteases that facilitate tumor cells proliferation, angiogenesis, invasion, and metastasis. This review summarizes the current knowledge about the mechanisms of thrombin-mediated proinflammatory responses in cancer pathology for a better understanding and hence a better management of this disease.
      PubDate: 2017-03-01T10:45:23.21184-05:0
      DOI: 10.1002/jcp.25753
       
  • Impact of Cryopreservation on Caprine Fetal Adnexa Derived Stem Cells and
           Its Evaluation for Growth Kinetics, Phenotypic Characterization, and Wound
           Healing Potential in Xenogenic Rat Model
    • Authors: Anjali Somal; Irfan A. Bhat, Indu B. , Anuj P. Singh, Bibhudatta S.K. Panda, Perumal A. Desingu, Sriti Pandey, Mukesh K. Bharti, Amar Pal, Guttula Saikumar, Vikash Chandra, Guttula Taru Sharma
      Abstract: This study was conducted to know the impact of cryopreservation on caprine fetal adnexa derived mesenchymal stem cells (MSCs) on the basic stem cell characteristics. Gravid caprine uteri (2–3 months) were collected from local abattoir to derive (amniotic fluid [cAF], amniotic sac [cAS], Wharton's jelly [cWJ], and cord blood [cCB]) MSCs and expanded in vitro. Cells were cryopreserved at 3rd passage (P3) using 10% DMSO. Post-thaw viability and cellular properties were assessed. Cells were expanded to determine growth kinetics, tri-lineage differentiation, localization, and molecular expression of MSCs and pluripotency markers; thereafter, these cells were transplanted in the full-thickness (2 × 2cm2) rat skin wound to determine their wound healing potential. The post-thaw (pt) growth kinetics study suggested that cWJ MSCs expanded more rapidly with faster population doubling time (PDT) than that of other fetal adnexa MSCs. The relative mRNA expression of surface antigens (CD73, CD90, and CD 105) and pluripotency markers (Oct4, KLF, and cMyc) was higher in cWJ MSCs in comparison to cAS, cAF, and cCB MSCs post-thaw. The percent wound contraction on 7th day was more than 50% for all the MSC-treated groups (pre and post-thaw), against 39.55% in the control group. On day 28th, 99% and more wound contraction was observed in cAF, cAF-pt, cAS-pt, cWJ, cWJ-pt, and cCB, MSCs with better scores for epithelization, neovascularization, and collagen characteristics at a non-significant level. It is concluded that these MSCs could be successfully cryopreserved without altering their stemness and wound healing properties. J. Cell. Physiol. 9999: 1–15, 2017. © 2016 Wiley Periodicals, Inc.The present study showed that cWJ MSCs have better pluripotency and post thaw survivability than other fetal adnexa derived MSCs and all the adnexa derived MSCs were found to heal excision wound faster and quality of healed wound was also better than control, indicating that these cells can be cryopreserved and may be used for regenerative cell therapy.
      PubDate: 2017-03-01T10:30:35.320275-05:
      DOI: 10.1002/jcp.25731
       
  • Notch Signaling Participates in TGF-β-Induced SOST Expression Under
           Intermittent Compressive Stress
    • Authors: Jeeranan Manokawinchoke; Piyamas Sumrejkanchanakij, Prasit Pavasant, Thanaphum Osathanon
      Abstract: Notch signaling is regulated by mechanical stimuli in various cell types. It has previously been reported that intermittent compressive stimuli enhanced sclerostin (SOST) expression in human periodontal ligament cells (hPDLs) by regulating transforming growth factor-β (TGF-β) expression. The aim of the present study was to determine the involvement of Notch signaling in the TGF-β-induced SOST expression in hPDLs. Cells were treated with intermittent compressive stress in a computer-controlled apparatus for 24 h. The mRNA and protein expression of the cells were determined by real-time polymerase chain reaction and Western blot analysis, respectively. In some experiments, the target signaling pathway was impeded by the addition of a TGF-β receptor kinase inhibitor (SB431542) or a γ-secretase inhibitor (DAPT). The results demonstrated that hPDLs under intermittent compressive stress exhibited significantly higher NOTCH2, NOTCH3, HES1, and HEY1 mRNA expression compared with control, indicating that mechanical stress induced Notch signaling. DAPT pretreatment markedly reduced the intermittent stress-induced SOST expression. The expression of NOTCH2, NOTCH3, HES1, and HEY1 mRNA under compressive stress was significantly reduced after pretreatment with SB431542, coinciding with a reduction in SOST expression. Recombinant human TGF-β1 enhanced SOST, Notch receptor, and target gene expression in hPDLs. Further, DAPT treatment attenuated rhTGF-β1-induced SOST expression. In summary, intermittent compressive stress regulates Notch receptor and target gene expression via the TGF-β signaling pathway. In addition, Notch signaling participates in TGF-β-induced SOST expression in hPDLs. J. Cell. Physiol. 9999: 1–10, 2016. © 2016 Wiley Periodicals, Inc.Intermittent stress stimulates TGF-β signaling via an unknown mechanism. TGF-β, in turn, induces the expression of SOST, NOTCH2, NOTCH3, HES1, and HEY1, whose expression was reduced by SB431542. Inhibition SOST expression, but not that of TGF-β by DAPT and NOTCH2 shRNA indicate that NOTCH signaling is a downstream participant in the TGF-β-induced SOST expression.
      PubDate: 2017-03-01T09:26:05.50402-05:0
      DOI: 10.1002/jcp.25740
       
  • Tolerogenic probiotics: potential immunoregulators in Systemic Lupus
           Erythematosus
    • Authors: Seyed-Alireza Esmaeili; Mahmoud Mahmoudi, Amir Abbas Momtazi, Amirhossein Sahebkar, Hassan Doulabi, Maryam Rastin
      Abstract: Probiotics are commensal or nonpathogenic microbes that colonize the gastrointestinal tract and confer beneficial effects on the host through several mechanisms such as competitive exclusion, anti-bacterial effects, and modulation of immune responses. There is growing evidence supporting the immunomodulatory ability of some probiotics. Several experimental and clinical studies have been shown beneficial effect of some probiotic bacteria, particularly Lactobacillus and Bifidobacteria strains, on inflammatory and autoimmune diseases. Systemic lupus erythematosus (SLE) is an autoimmune disease that is mainly characterized by immune intolerance towards self-antigens. Some immunomodulatory probiotics have been found to regulate immune responses via tolerogenic mechanisms. Dendritic and T regulatory (Treg) cells, IL-6, IFN-γ, IL-17, and IL-23 can be considered as the most determinant dysregulated mediators in tolerogenic status. As demonstrated by documented experimental and clinical trials on inflammatory and autoimmune diseases, a number of probiotic bacterial strains can restore tolerance in host through modification of such dysregulated mediators. Since there are limited reports regarding to impact of probiotic supplementation in SLE patients, the preset review was aimed to suggest a number of probiotics bacteria, mainly from Bifidobacteria and Lactobacillus strains that are able to ameliorate immune responses. The aim was followed through literature survey on immunoregulatory probiotics that can restore tolerance and also modulate the important dysregulated pro/anti-inflammatory cytokines contributing to the pathogenesis of SLE.Since there are limited reports regarding to impact of probiotic supplementation in SLE patients, the preset review was aimed to suggest a number of probiotics bacteria, mainly from Bifidobacteria and Lactobacillus strains that are able to ameliorate immune responses. The aim was followed through literature survey on immunoregulatory probiotics that can restore tolerance and also modulate the important dysregulated pro/anti-inflammatory cytokines contributing to the pathogenesis of SLE.
      PubDate: 2017-03-01T09:20:36.037057-05:
      DOI: 10.1002/jcp.25748
       
  • Uremic Serum Impairs Osteogenic Differentiation of Human Bone Marrow
           Mesenchymal Stromal Cells
    • Authors: Elena Della Bella; Stefania Pagani, Gianluca Giavaresi, Irene Capelli, Giorgia Comai, Chiara Donadei, Maria Cappuccilli, Gaetano La Manna, Milena Fini
      Abstract: Chronic Kidney Disease-Mineral and Bone Disorder (CKD-MBD) is characterized by an increased fracture risk. Bone marrow mesenchymal stromal cells (BMSCs) may be involved in the pathogenesis of bone disease and, in view of their promising potential applications in bone tissue engineering, the effect of uremia on BMSCs regenerative potential represents a central issue. The present study evaluated in vitro the effect of a serum pool from hemodialysis patients on BMSCs to observe its influence on osteogenic differentiation. Besides alterations in spatial organization and cytotoxicity along with hyperproliferation, gene expression analysis suggested an impairment in the osteogenic differentiation. More importantly, Receptor activator of nuclear factor kappa-B ligand (RANKL) was upregulated with a mild reduction in osteoprotegerin levels. In summary, uremic environment seems to impair BMSCs osteogenic differentiation. Moreover BMSCs themselves may enhance osteoclastogenesis, feasibly contributing to the altered bone remodeling in CKD-MBD patients. J. Cell. Physiol. 9999: 1–9, 2016. © 2016 Wiley Periodicals, Inc.BMSCs were cultured in presence of a serum pool from hemodialysis patients (HUS) to study the effect of uremia on BMSC behavior. HUS induced both hyperproliferation and cytotoxicity, with an alteration in the spatial organization of cells. Moreover, osteogenic differentiation was impaired: a reduced mineralization and an altered pattern of gene expression were observed, in particular upregulation of RANKL and reduction in OPG levels.
      PubDate: 2017-03-01T09:10:30.160074-05:
      DOI: 10.1002/jcp.25732
       
  • Dlx3 and GCM-1 functionally coordinate the regulation of placental growth
           factor in human trophoblast-derived cells
    • Authors: Sha Li; Mark S. Roberson
      Abstract: Placental growth factor (PGF) is abundantly expressed by trophoblast cells within human placentae and is important for trophoblast development and placental vascularization. Circulating maternal serum levels of PGF are dynamically upregulated across gestation in normal pregnancies, whereas low circulating levels and placental production of PGF have been implicated in the pathogenesis of preeclampsia and other gestational diseases. However, the underlying molecular mechanism of regulating PGF expression in the human placenta remains poorly understood. In this study, we demonstrated that transcription factors Distal-less 3 (DLX3) and Glial cell missing-1 (GCM1) were both sufficient and required for PGF expression in human trophoblast-derived cells by overexpression and knockdown approaches. Surprisingly, while DLX3 and GCM1 were both positive regulators of PGF, co-overexpression of DLX3 and GCM1 led to an antagonist effect on PGF expression on the endogenous gene and a luciferase reporter. Further, deletion and site-directed mutagenesis studies identified a novel regulatory element on the PGF promoter mediating both DLX3- and GCM1-dependent PGF expression. This regulatory region was also found to be essential for the basal activity of the PGF promoter. Finally, Chromatin-immunoprecipitation (ChIP) assays revealed colocalization of DLX3 and GCM1 at the identified regulatory region on the PGF promoter. Taken together, our studies provide important insights into intrinsic regulation of human placental PGF expression through the functional coordination of DLX3 and GCM1, and are likely to further the understanding of pathogenesis of PGF dysregulation in preeclampsia and other disease conditions.Our studies examine the role and requirement for the transcription factors Dlx3 and GCM-1 in the regulation of placental growth factor (PGF) in human placental trophoblast cell lines. Extensive over-expression and knockdown studies reveal a requirement for both factors; however, the combined action of Dlx3 and GCM-1 is antagonistic to PGF expression. Our studies identify a novel cis-acting regulatory element in the proximal region of the PGF gene promoter that binds both factors and is required for the antagonist effects of Dlx3 on GCM-1.
      PubDate: 2017-03-01T09:05:27.51505-05:0
      DOI: 10.1002/jcp.25752
       
  • Integrated Analysis and MicroRNA Expression Profiling Identified Seven
           miRNAs Associated With Progression of Oral Squamous Cell Carcinoma
    • Authors: Zhong-Yi Yan; Zhi-Qing Luo, Lai-Jian Zhang, Jia Li, Jia-Qiang Liu
      Abstract: MicroRNAs have been used as diagnostic and prognostic biomarkers for many cancers including oral squamous cell carcinoma (OSCC). Several studies have been shown that microRNA (miRNA) play important roles during the progression of OSCC. However, the results vary largely in different studies due to different platforms and sample sizes. In this study, we systematically evaluated a large scale of miRNA profiles from current qualified OSCC samples, and further investigated the functions of genes regulated by these key miRNAs as well as the signaling pathways through which these miRNA effect carcinogenesis. Seven key miRNAs were identified, and of which three were significantly upregulated, including hsa-miR-21, hsa-miR-31, hsa-miR-338, and four were downregulated, namely hsa-miR-125b, hsa-miR-133a, hsa-miR-133b, and hsa-miR-139. The function enrichment analysis revealed that target genes of upregulated miRNAs were associated with cellular protein metabolic process, macromolecule metabolic process, and TGF-beta pathway, while the targets of downregulated were enriched in negative regulation of macromolecule biosynthetic process and gene expression, and p53, long-term potentiation and adherens junction pathways. Transcription factor analysis revealed that there were 67 (51.1%) transcription factors influenced by both up and downregulated miRNAs. In summary, seven key miRNAs were found to play essential role in progression of OSCC, as well as the target genes and transcription factors of these miRNAs. The potential functions of these target genes identified in our study may be profitable to diagnosis and prognostic prediction of OSCC as biomarkers. J. Cell. Physiol. 9999: 1–8, 2016. © 2016 Wiley Periodicals, Inc.Seven key miRNAs were found to play essential role in progression of OSCC, as well as the target genes and transcription factors of these miRNAs. The potential functions of these target genes identified in our study may be profitable to diagnosis and prognostic prediction of OSCC as biomarkers.
      PubDate: 2017-03-01T09:00:38.977533-05:
      DOI: 10.1002/jcp.25728
       
  • Hyaluronic Acid Nanohydrogel Loaded With Quercetin Alone or in Combination
           to a Macrolide Derivative of Rapamycin RAD001 (Everolimus) as a New
           Treatment for Hormone-Responsive Human Breast Cancer
    • Authors: Vincenzo Quagliariello; Rosario Vincenzo Iaffaioli, Emilia Armenia, Ottavia Clemente, Manlio Barbarisi, Guglielmo Nasti, Massimiliano Berretta, Alessandro Ottaiano, Alfonso Barbarisi
      Abstract: The aim of this study is based on the evaluation of anticancer, anti-inflammatory activities, and cellular uptake of hyaluronic acid nanohydrogel of quercetin tested alone and in combination to a macrolide derivative of rapamycin RAD001 (everolimus) on hormone-responsive breast cancer cell line MCF-7. Biological investigations were focused on the receptor mediated cellular internalization of the nanohydrogel and its abilities to reduce secretion of several cytokines (IL-8, IL-6, IL-19), VEGF, and metalloproteases (MMP-2, MMP-9) under pro-inflammatory conditions. Nanohydrogel show a CD44 dependent endocytosis with evident time dependent cytoplasmatic accumulation with abilities to reduce secretion of all cytokines of ∼60% compared to untreated cells. Combination of formulated quercetin and everolimus leads to a synergistic cytotoxic effects with a Combination Index of 0.38. These results highlights the importance of synergistic effect of the hyaluronic acid nanohydrogel of quercetin with everolimus in the regulation of human breast cancer cell proliferation and emphasize the antitumor and anti-inflammatory properties of the nanocarrier. J. Cell. Physiol. 9999: 1–12, 2016. © 2016 Wiley Periodicals, Inc.Hyaluronic acid nanohydrogel of Quercetin has anti-anticancer and antinflammatory activity in human breast cancer cells. Combination of Everolimus and hyaluronic acid nanohydrogel of Quercetin leads to synergistic cytotoxicity in MCF-7 call line.
      PubDate: 2017-03-01T08:55:25.407896-05:
      DOI: 10.1002/jcp.25587
       
  • Plasma Membrane Calcium ATPase 4 (PMCA4) co-ordinates Calcium and Nitric
           Oxide signaling in regulating Murine Sperm functional activity
    • Authors: Kristine E. Olli; Kun Li, Deni S. Galileo, Patricia A. Martin-DeLeon
      Abstract: Reduced sperm motility (asthenospermia) and resulting infertility arise from deletion of the Plasma Membrane Ca2+-ATPase 4 (Pmca4) gene which encodes the highly conserved Ca2+ efflux pump, PMCA4. This is the major Ca2+ clearance protein in murine sperm. Since the mechanism underlying asthenospermia in PMCA4's absence or reduced activity is unknown, we investigated if sperm PMCA4 negatively regulates nitric oxide synthases (NOSs) and when absent NO, peroxynitrite, and oxidative stress levels are increased. Using co-immunoprecipitation (Co-IP) and Fluorescence Resonance Energy Transfer (FRET), we show an association of PMCA4 with the NOSs in elevated cytosolic [Ca2+] in capacitated and Ca2+ ionophore-treated sperm and with neuronal (nNOS) at basal [Ca2+] (ucapacitated sperm). FRET efficiencies for PMCA4-eNOS were 35% and 23% in capacitated and uncapacitated sperm, significantly (P 
      PubDate: 2017-03-01T07:55:27.125042-05:
      DOI: 10.1002/jcp.25882
       
  • TGFβ Signalling and the Control of Myofibroblast Differentiation:
           Implications for Chronic Inflammatory Disorders
    • Authors: Jonathon M Carthy
      Abstract: The myofibroblast is a highly specialized cell type that plays a critical role during normal tissue wound healing, but also contributes pathologically to chronic inflammatory conditions such as fibrosis and cancer. As fibrotic conditions continue to be a major burden to the public health system, novel therapies that target the function of myofibroblasts may show promise in the clinic. The cytokine transforming growth factor β (TGFβ) is the most potent known inducer of myofibroblast differentiation and thus represents a powerful target to modify myofibroblast function during disease. This review focuses on our current understanding of the key signaling pathways activated by TGFβ during myofibroblast differentiation. This article is protected by copyright. All rights reserved
      PubDate: 2017-03-01T07:50:39.648093-05:
      DOI: 10.1002/jcp.25879
       
  • GM13133 Is a Negative Regulator in Mouse White Adipocytes Differentiation
           and Drives the Characteristics of Brown Adipocytes
    • Authors: LiangHui You; YaHui Zhou, XianWei Cui, XingYun Wang, YaZhou Sun, Yao Gao, Xing Wang, Juan Wen, Kaipeng Xie, RanRan Tang, ChenBo Ji, XiRong Guo
      Abstract: Obesity is tightly associated with the disturbance of white adipose tissue storing excess energy. Thermogenic adipocytes (brown and beige) exert a critical role of oxidizing nutrients at the high rates through non-shivering thermogenesis. The recruitment of brown characteristics in white adipocytes, termed browning, has been considered as a promising strategy for treating obesity and associated metabolic complications. Recently, long noncoding RNAs play a crucial role in regulating tissue development and participating in disease pathogenesis, yet their effects on the conversion of white into brown-like adipocytes and thermogenic function were not totally understood. Here, we identified a mouse brown adipose specific expressed lncRNA, termed GM13133. Moreover, a considerable amount of GM13133 is expressed in adipocytes and actively modulated by cold, β3-adrenergic agonist and cAMP stimuli, implying a potential role in the conversion from white to brown adipocytes. Overexpression of GM13133 didn't affect the proliferation of mouse white pre-adipocytes, but inhibited white adipocyte differentiation by decreasing lipid accumulation. The forced expression of GM13133 also significantly drove the conversion of white into brown-like adipocytes with the enhanced mitochondrial biogenesis and the induced expression of brown adipocytes specific markers. A global mRNA analysis further indicated the possible regulatory role of cAMP signaling pathway in GM13133 mediated white-to-brown adipocytes conversion. Our results identified a lncRNA-mediated modulation in primary mouse white adipocyte differentiation and indicate the functional significance of GM13133 in promoting browning of white adipocytes and maintenance of thermogenesis, further providing a potential strategy to treating obesity. This article is protected by copyright. All rights reserved
      PubDate: 2017-03-01T07:50:38.368695-05:
      DOI: 10.1002/jcp.25878
       
  • Identification of Murine Phosphodiesterase 5A Isoforms and their
           Functional Characterization in HL-1 Cardiac Cell Line
    • Authors: Federica Campolo; Alessandra Zevini, Silvia Cardarelli, Lucia Monaco, Federica Barbagallo, Manuela Pellegrini, Marisa Cornacchione, Antonio Di Grazia, Valeria De Arcangelis, Daniele Gianfrilli, Mauro Giorgi, Andrea Lenzi, Andrea M. Isidori, Fabio Naro
      Abstract: Phosphodiesterase 5A (PDE5A) specifically degrades the ubiquitous second messenger cGMP and experimental and clinical data highlight its important role in cardiac diseases. To address PDE5A role in cardiac physiology, three splice variants of the PDE5A were cloned for the first time from mouse cDNA library (mPde5a1, mPde5a2 and mPde5a3). The predicted amino acidic sequences of the three murine isoforms are different in the N-terminal regulatory domain. mPDE5A isoforms were transfected in HEK293T cells and they showed high affinity for cGMP and similar sensitivity to sildenafil inhibition. RT-PCR analysis showed that mPde5a1, mPde5a2 and mPde5a3 had differential tissue distribution. In the adult heart, mPde5a1 and mPde5a2 were expressed at different levels whereas mPde5a3 was undetectable. Overexpression of mPDE5As induced an increase of HL-1 number cells which progress into cell cycle. mPDE5A1 and mPDE5A3 overexpression increased the number of polyploid and binucleated cells, mPDE5A3 widened HL-1 areas and modulated hypertrophic markers more efficiently respect to the other mPDE5A isoforms. Moreover, mPDE5A isoforms had differential subcellular localization: mPDE5A1 was mainly localized in the cytoplasm, mPDE5A2 and mPDE5A3 were also nuclear localized. These results demonstrate for the first time the existence of three PDE5A isoforms in mouse and highlight their potential role in the induction of hypertrophy. This article is protected by copyright. All rights reserved
      PubDate: 2017-03-01T07:50:30.854052-05:
      DOI: 10.1002/jcp.25880
       
  • Steady State Peripheral Blood Provides Cells with Functional and Metabolic
           Characteristics of Real Hematopoietic Stem Cells
    • Authors: A. Bourdieu; M. Avalon, V. Lapostolle, S. Ismail, M. Mombled, C. Debeissat, M. Guérinet, P. Duchez, J. Chevaleyre, M. Vlaski-Lafarge, A. Villacreces, V. Praloran, Z. Ivanovic, P. Brunet de la Grange
      Abstract: Hematopoietic stem cells (HSCs), which are located in the bone marrow, also circulate in cord and peripheral blood. Despite high availability, HSCs from steady state peripheral blood (SSPB) are little known and not used for research or cell therapy. We thus aimed to characterize and select HSCs from SSPB by a direct approach with a view to delineating their main functional and metabolic properties and the mechanisms responsible for their maintenance. We chose to work on Side Population (SP) cells which are highly enriched in HSCs in mouse, human bone marrow, and cord blood. However, no SP cells from SSBP have as yet been characterized. Here we showed that SP cells from SSPB exhibited a higher proliferative capacity and generated more clonogenic progenitors than non-SP cells in vitro. Furthermore, xenotransplantation studies on immunodeficient mice demonstrated that SP cells are up to 45 times more enriched in cells with engraftment capacity than non-SP cells. From a cell regulation point of view, we showed that SP activity depended on O2 concentrations close to those found in HSC niches, an effect which is dependent on both hypoxia-induced factors HIF-1α and HIF-2α. Moreover SP cells displayed a reduced mitochondrial mass and, in particular, a lower mitochondrial activity compared to non-SP cells, while they exhibited a similar level of glucose incorporation. These results provided evidence that SP cells from SSPB displayed properties of very primitive cells and HSC, thus rendering them an interesting model for research and cell therapy. This article is protected by copyright. All rights reserved
      PubDate: 2017-03-01T07:50:24.289104-05:
      DOI: 10.1002/jcp.25881
       
  • Anti-CRISPR Proteins: Counterattack of Phages on Bacterial Defense
           (CRISPR/Cas) System
    • Authors: Kulbhushan Chaudhary; Anirudha Chattopadhyay, Dharmendra Pratap
      Abstract: Since the dawn of life there is a never ending strife between bacteria and phages. Both are perpetually changing their strategies to take over each other. CRISPR/Cas is the most widespread defense system used by bacteria against mobile genetic elements (MGEs) such as phages, cojugative palsmids, transoposons and pathogenicity islands. This system utilizes small guide RNA molecules to protect against phages infection and invasion by MGEs. Phages circumvent to these antiviral barriers by point mutation in PAM (protospacer-adjacent motif) sequence, genome rearrangements and by using anti-CRISPR proteins. This article is protected by copyright. All rights reserved
      PubDate: 2017-03-01T07:45:29.60748-05:0
      DOI: 10.1002/jcp.25877
       
  • Quantification of Exosomes
    • Authors: Erik H. Koritzinsky; Jonathan M. Street, Robert A. Star, Peter S.T. Yuen
      Abstract: Exosomes are released by cells as self-contained vesicles with an intact lipid bilayer that encapsulates a small portion of the parent cell. Exosomes have been studied widely as information-rich sources of potential biomarkers that can reveal cellular physiology. We suggest that quantification is essential to understand basic biological relationships between exosomes and their parent cells and hence the underlying interpretation of exosome signals. The number of methods for quantifying exosomes has expanded as interest in exosomes has increased. However, a consensus on proper quantification has not developed, making each study difficult to compare to another. Overcoming this ad hoc approach will require widely available standards that have been adequately characterized, and multiple comparative studies across platforms. We outline the current status of these technical approaches and our view of how they can become more coherent. J. Cell. Physiol. 9999: 1–4, 2016. © 2016 Wiley Periodicals, Inc.As exosomes are released from cells, they represent not only a rich source of biomarkers, but also a potential source of insight into parent cell physiology. Quantitation of exosomes is essential for progress in both of these areas. Several methods have been developed to quantitate exosomes, and we summarize the current status and future directions toward standardization of these methods.
      PubDate: 2017-03-01T07:02:02.139595-05:
      DOI: 10.1002/jcp.25387
       
  • Celastrol Attenuates Cadmium-Induced Neuronal Apoptosis via Inhibiting
           Ca2+-CaMKII-Dependent Akt/mTOR Pathway
    • Authors: Ruijie Zhang; Yu Zhu, Xiaoqing Dong, Beibei Liu, Nana Zhang, Xiaoxue Wang, Lei Liu, Chong Xu, Shile Huang, Long Chen
      Abstract: Cadmium (Cd), an environmental and industrial pollutant, affects the nervous system and consequential neurodegenerative disorders. Recently, we have shown that celastrol prevents Cd-induced neuronal cell death partially by suppressing Akt/mTOR pathway. However, the underlying mechanism remains to be elucidated. Here, we show that celastrol attenuated Cd-elevated intracellular-free calcium ([Ca2+]i) level and apoptosis in neuronal cells. Celastrol prevented Cd-induced neuronal apoptosis by inhibiting Akt-mediated mTOR pathway, as inhibition of Akt with Akt inhibitor X or ectopic expression of dominant negative Akt reinforced celastrol's prevention of Cd-induced phosphorylation of S6K1/4E-BP1 and cell apoptosis. Furthermore, chelating intracellular Ca2+ with BAPTA/AM or preventing [Ca2+]i elevation using EGTA potentiated celastrol's repression of Cd-induced [Ca2+]i elevation and consequential activation of Akt/mTOR pathway and cell apoptosis. Moreover, celastrol blocked Cd-elicited phosphorylation of CaMKII, and pretreatment with BAPTA/AM or EGTA enhanced celastrol's suppression of Cd-increased phosphorylation of CaMKII in neuronal cells, implying that celastrol hinders [Ca2+]i-mediated CaMKII phosphorylation. Inhibiting CaMKII with KN93 or silencing CaMKII attenuated Cd activation of Akt/mTOR pathway and cell apoptosis, and this was strengthened by celastrol. Taken together, these data demonstrate that celastrol attenuates Cd-induced neuronal apoptosis via inhibiting Ca2+-CaMKII-dependent Akt/mTOR pathway. Our findings underscore that celastrol may act as a neuroprotective agent for the prevention of Cd-induced neurodegenerative disorders. J. Cell. Physiol. 9999: 1–13, 2016. © 2016 Wiley Periodicals, Inc.Celastrol prevents Cd-induced elevation of [Ca2+]i, which results in inhibition of CaMKII phosphorylation. This leads to suppression of CaMKII-dependent Akt/mTOR pathway, thereby preventing Cd-induced apoptosis in neuronal cells.
      PubDate: 2017-02-28T09:25:30.042774-05:
      DOI: 10.1002/jcp.25703
       
  • Versatile Roles of Intracellularly Located TRPV1 Channel
    • Authors: Rui Zhao; Suk Ying Tsang
      Abstract: The ubiquitous expression in many organs throughout the body and the ability to respond to a wide variety of physical and chemical stimuli have brought transient receptor potential (TRP) channels to the vanguards of our sensory systems. TRP vanilloid-1 (TRPV1) is the founding member of the TRPV subfamily. TRPV1 can be activated by noxious heat, protons, and vanilloids. Previous studies have shown that TRPV1 is located on the plasma membrane, serving to non-selectively permeate calcium ion from the extracellular region to the cytoplasm. Interestingly, increasing evidence suggests that TRPV1 is also located intracellularly in various cell types such as neurons, myocytes, and numerous cancer cells. By immunocytochemistry and/or fractionation followed by Western blotting, TRPV1 was found to express on the endoplasmic reticulum/sarcoplasmic reticulum and the mitochondria. By using various pharmacological and molecular tools, intracellular TRPV1 was also found to functionally express to control calcium level both inside the organelles and in the cytoplasm. Recent studies have shown that intracellularly located TRPV1 serves versatile functions in various physiological and pathological conditions (e.g., exercise endurance and hypertrophy). In this review, we not only have summarized the well-characterized roles of TRPV1, but also have highlighted the increasing importance of intracellular TRPV1-mediated pathways. Lastly, we have pointed out future research direction for answering several important questions that have remained unanswered. Vigorous investigation of the emerging roles of intracellular TRPV1 can allow a better understanding of how TRPV1 controls the cellular calcium homeostasis and its role in various physiological and pathophysiological conditions. J. Cell. Physiol. 9999: 1–9, 2016. © 2016 Wiley Periodicals, Inc.Increasing evidence suggests that TRPV1 is located on the endoplasmic reticulum/sarcoplasmic reticulum and on the mitochondria. Intracellular TRPV1 was found to functionally express to control calcium level both inside the organelles and in the cytoplasm and serve versatile functions in various physiological and pathological conditions. In this review, we not only have summarized the well-characterized roles of TRPV1, but also have highlighted the increasing importance of intracellular TRPV1-mediated pathways.
      PubDate: 2017-02-28T09:15:26.746156-05:
      DOI: 10.1002/jcp.25704
       
  • Epigenetics in Reactive and Reparative Cardiac Fibrogenesis: The Promise
           of Epigenetic Therapy
    • Authors: Asish K. Ghosh; Rahul Rai, Panagiotis Flevaris, Douglas E. Vaughan
      Abstract: Epigenetic changes play a pivotal role in the development of a wide spectrum of human diseases including cardiovascular diseases, cancer, diabetes, and intellectual disabilities. Cardiac fibrogenesis is a common pathophysiological process seen during chronic and stress-induced accelerated cardiac aging. While adequate production of extracellular matrix (ECM) proteins is necessary for post-injury wound healing, excessive synthesis and accumulation of extracellular matrix protein in the stressed or injured hearts causes decreased or loss of lusitropy that leads to cardiac failure. This self-perpetuating deposition of collagen and other matrix proteins eventually alter cellular homeostasis; impair tissue elasticity and leads to multi-organ failure, as seen during pathogenesis of cardiovascular diseases, chronic kidney diseases, cirrhosis, idiopathic pulmonary fibrosis, and scleroderma. In the last 25 years, multiple studies have investigated the molecular basis of organ fibrosis and highlighted its multi-factorial genetic, epigenetic, and environmental regulation. In this minireview, we focus on five major epigenetic regulators and discuss their central role in cardiac fibrogenesis. Additionally, we compare and contrast the epigenetic regulation of hypertension-induced reactive fibrogenesis and myocardial infarction-induced reparative or replacement cardiac fibrogenesis. As microRNAs—one of the major epigenetic regulators—circulate in plasma, we also advocate their potential diagnostic role in cardiac fibrosis. Lastly, we discuss the evolution of novel epigenetic-regulating drugs and predict their clinical role in the suppression of pathological cardiac remodeling, cardiac aging, and heart failure. J. Cell. Physiol. 9999: 1–16, 2017. © 2016 Wiley Periodicals, Inc.In this minireview, we discuss epigenetic regulation of cardiac fibrogenesis. Cardiac vascular occlusion-induced myocardial infarction and sustained high blood pressure alter the levels and activities of different epigenetic regulators that lead to epigenetic modifications of chromatin histones and transcription factors and non-physiological matrix protein synthesis. Normalization of epigenetic modifications targeting altered specific epigenetic regulator may be an ideal therapeutic approach to control cardiac fibrogenesis.
      PubDate: 2017-02-28T08:25:27.782287-05:
      DOI: 10.1002/jcp.25699
       
  • Human Telomerase Reverse Transcriptase (hTERT) Positively Regulates 26S
           Proteasome Activity
    • Authors: Eunju Im; Jong Bok Yoon, Han-Woong Lee, Kwang Chul Chung
      Abstract: Human telomerase reverse transcriptase (hTERT) is the catalytic subunit of telomerase, an RNA-dependent DNA polymerase that elongates telomeric DNA. hTERT displays several extra-telomeric functions that are independent of its telomere-regulatory function, including tumor progression, and neuronal cell death regulation. In this study, we evaluated these additional hTERT non-telomeric functions. We determined that hTERT interacts with several 19S and 20S proteasome subunits. The 19S regulatory particle and 20S core particle are part of 26S proteasome complex, which plays a central role in ubiquitin-dependent proteolysis. In addition, hTERT positively regulated 26S proteasome activity independent of its enzymatic activity. Moreover, hTERT enhanced subunit interactions, which may underlie hTERT's ability of hTERT to stimulate the 26S proteasome. Furthermore, hTERT displayed cytoprotective effect against ER stress via the activation of 26S proteasome in acute myeloid leukemia cells. Our data suggest that hTERT acts as a novel chaperone to promote 26S proteasome assembly and maintenance. J. Cell. Physiol. 9999: 1–11, 2016. © 2016 Wiley Periodicals, Inc.The present study demonstrated that hTERT positively regulates 26S proteasome activity independent of its enzymatic activity. Moreover, hTERT displayed cytoprotective effect against ER stress via the activation of 26S proteasome in acute myeloid leukemia cells. Our data suggest that hTERT acts as a novel chaperone to promote 26S proteasome assembly and maintenance.
      PubDate: 2017-02-28T08:10:29.642961-05:
      DOI: 10.1002/jcp.25607
       
  • Chemoresistance of lung and breast cancer cells growing under prolonged
           periods of serum starvation
    • Authors: Juan Sebastian Yakisich; Rajkumar Venkatadri, Neelam Azad, Anand Krishnan V. Iyer
      Abstract: The efficacy of chemotherapy is hindered by both tumor heterogeneity and acquired or intrinsic multi-drug resistance caused by the contribution of multidrug resistance proteins and stemness-associated prosurvival markers. Therefore, targeting multi-drug resistant cells would be much more effective against cancer. In this study, we characterized the chemoresistance properties of adherent (anchorage-dependent) lung H460 and breast MCF-7 cancer cells growing under prolonged periods of serum starvation (PPSS). We found that under PPSS, both cell lines were highly resistant to Paclitaxel, Colchicine, Hydroxyurea, Obatoclax, Wortmannin, and LY294002. Levels of several proteins associated with increased stemness such as Sox2, MDR1, ABCG2, and Bcl-2 were found to be elevated in H460 cells but not in MCF-7 cells. While pharmacological inhibition of either MDR1, ABCG2, Bcl-2 with Verapamil, Sorafenib, or Obatoclax, respectively decreased the levels of their target proteins under routine culture conditions as expected, such inhibition did not reverse PX resistance in PPSS conditions. Paradoxically, treatment with inhibitors in serum-starved conditions produced an elevation of their respective target proteins. In addition, we found that Digitoxin, an FDA approved drug that decrease the viability of cancer cells growing under PPSS, downregulates the expression of Sox2, MDR1, phospho- AKT, Wnt5a/b, and β-catenin. Our data suggest that PPSS-induced chemoresistance is the result of extensive rewiring of intracellular signaling networks and that multi-resistance can be effectively overcome by simultaneously targeting multiple targets of the rewired network. Furthermore, our PPSS model provides a simple and useful tool to screen drugs for their ability to target multiple pathways of cancer resistance. J. Cell. Physiol. 9999: 1–11, 2016. © 2016 Wiley Periodicals, Inc.Cancer cells growing under prolonged periods of serum starvation (PPSS) become multidrug resistant. Under PPSS cancer cells respond aberrantly and paradoxically to some anticancer drugs likely by rewire signaling pathway networks.
      PubDate: 2017-02-28T07:50:54.531744-05:
      DOI: 10.1002/jcp.25514
       
  • Dedicator of cytokinesis 2 in cell signaling regulation and disease
           development
    • Authors: Xia Guo; Shi-You Chen
      Abstract: Dedicator of cytokinesis 2 (DOCK2) is a CDM family protein containing DOCK homology region (DHR)-1 and DHR-2, Src-homology 3 (SH3) domain, and C-terminal polybasic amino acid cluster. The CDM family consists of 11 mammalian members and is classified into four subfamilies, the DOCK-A, -B, -C, and -D. DOCK2 is a member of DOCK-A subfamily and an atypical guanine exchange factor regulating the loading of GTP to activate Rac. It is primarily found in peripheral blood, spleen, and thymus and mainly expressed in lymphocytes and macrophages of various organs. DOCK2 is also expressed in microglial in brain and is induced in neointima smooth muscle following vascular injury. Functionally, DOCK2 is involved in cell motility, polarity, adhesion, proliferation, and apoptosis. It is essential for lymphocyte migration and activation as well as neutrophil chemotaxis. DOCK2 also regulates the differentiation of natural killer T cells, type 2 T helper cells, and plasmacytoid dendritic cells. In addition, it is important for the growth of B cell lymphoma and prostate cancer cells. Deletion of DOCK2 enables long-term cardiac allograft survival. Moreover, DOCK2 is associated with the Alzheimer Disease, HIV development, and the early-onset of invasive infections. Recently, we found that DOCK2 plays a critical role in SMC phenotypic modulation and vascular remodeling. In this review, we will briefly summarize recent advancement of DOCK2 function. J. Cell. Physiol. 9999: 1–10, 2016. © 2016 Wiley Periodicals, Inc.Dedicator of cytokinesis 2 (DOCK2) is an atypical guanine exchange factor regulating the loading of GTP to activate Rac. It is a very important protein factor involved in multiple cellular processes including cell motility, polarity, adhesion, proliferation, and apoptosis. Most studies have been focused on its regulation on immune cells. We recently reported that DOCK2 regulates vascular smooth muscle cell phenotypic modulation.
      PubDate: 2017-02-28T07:35:45.066974-05:
      DOI: 10.1002/jcp.25512
       
  • Isolation, identification, and characterization of cancer stem cells: A
           review
    • Authors: Mohammad Reza Abbaszadegan; Vahid Bagheri, Mahya Shariat Razavi, Amir Abbas Momtazi, Amirhossein Sahebkar, Mehran Gholamin
      Abstract: Cancer stem cells (CSCs) or tumor-initiating cells (TICs) as a small subset of neoplastic cells are able to produce a tumor (tumorigenesis), maintain the population of tumorigenic cells (self-renewal), and generate the heterogeneous cells constructing the entire tumor (pluripotency). The research on stationary and circulating CSCs due to resistance to conventional therapies and inability in complete eradication of cancer is critical for developing novel therapeutic strategies for a more effective reduction in the risk of tumor metastasis and cancer recurrence. This review compiles information about different methods of detection and dissociation, side population, cellular markers, and establishment culture of CSCs, as well as characteristics of CSCs such as tumorigenicity, and signaling pathways associated with self-renewal and the capability of the same histological tumor regeneration in various cancers.This review compiles information about detection, dissociation, side population, cellular markers, and establishment culture of cancer stem cells, as well as characteristics of CSCs such as tumorigenicity, signaling pathways associated with self-renewal and the capability of the same histological tumor regeneration in various cancers.
      PubDate: 2017-02-28T07:21:00.771459-05:
      DOI: 10.1002/jcp.25759
       
  • An Advanced Tri-culture Model to Evaluate the Dynamic Interplay Among
           Osteoblasts, Osteoclasts and Endothelial Cells
    • Authors: Stefania Pagani; Paola Torricelli, Francesca Veronesi, Francesca Salamanna, Simona Cepollaro, Milena Fini
      Abstract: The dynamic metabolism and the numerous roles of bone tissue necessitate a suitable in vitro model to represent them. In order to investigate the interaction among the several cell types composing bone microenvironment, we studied a tri-culture model including human osteoblasts (OBs), osteoclasts (OCs) and endothelial cells (HUVEC). While OBs are essential for bone deposition and OCs for bone resorption, the vasculature is necessary to provide growth factors, nutrients and oxygen in the mature tissue. The results of this study showed a strong mutual influence between OBs, OCs and HUVEC in term of proliferation, viability and activity (release of ALP, Coll I, OPG, RANKL, VEGF, CTSK, TGFβ and IL-6). The behavior of the single cultures demonstrated to be different compared to the bi- or tri-cultures and depending on the cell types involved: the coexistence of OBs and OCs stimulated the synthetic activity of both cell types, while the presence of HUVEC induced a stimulating role for OBs but mainly an inhibitory effect for OC. In addition, evidence of the effects of OBs and OCs on HUVEC is highlighted by their morphology: regular and able to “sketch” little vessels in presence of OBs, more disorganized and heterogeneous in presence of OCs.Taken together, these observations well characterize an advanced cellular model to be used as starting point for mimicking bone microenvironment in vivo, thus reducing the use of animals in the preclinical phase and offering a more reliable tool to test new and innovative biomaterials. This article is protected by copyright. All rights reserved
      PubDate: 2017-02-27T05:35:39.435191-05:
      DOI: 10.1002/jcp.25875
       
  • Melatonin-induced Increase of Lipid Droplets Accumulation and In Vitro
           Maturation in Porcine Oocytes Is Mediated by Mitochondrial Quiescence
    • Authors: Bin He; Chao Yin, Yabin Gong, Jie Liu, Huiduo Guo, Ruqian Zhao
      Abstract: Melatonin, the major pineal secretory product, has a significant impact on the female reproductive system. Recently, the beneficial effects of melatonin on mammalian oocyte maturation and embryonic development have drawn increased attention. However, the exact underlying mechanisms remain to be fully elucidated. This study demonstrates that supplementing melatonin to in vitro maturation (IVM) medium enhances IVM rate, lipid droplets (LDs) accumulation as well as triglyceride content in porcine oocytes. Decrease of mitochondrial membrane potential, mitochondrial respiratory chain complex IV activity as well as mitochondrial reactive oxygen species (mROS) content indicated that melatonin induced a decrease of mitochondrial activity. The copy number of mitochondrial DNA (mtDNA) which encodes essential subunits of oxidative phosphorylation (OXPHOS), was not affected by melatonin. However, the expression of mtDNA-encoded genes was significantly down-regulated after melatonin treatment. The DNA methyltransferase DNMT1, which regulates methylation and expression of mtDNA, was increased and translocated into the mitochondria in melatonin-treated oocytes. The inhibitory effect of melatonin on the expression of mtDNA was significantly prevented by simultaneous addition of DNMT1 inhibitor, which suggests that melatonin regulates the transcription of mtDNA through up-regulation of DNMT1 and mtDNA methylation. Increase of triglyceride contents after inhibition of OXPHOS indicated that mitochondrial quiescence is crucial for LDs accumulation in oocytes. Taken together, our results suggest that melatonin-induced reduction in mROS production and increase in IVM and LDs accumulation in porcine oocytes is mediated by mitochondrial quiescence. This article is protected by copyright. All rights reserved
      PubDate: 2017-02-27T05:35:33.507037-05:
      DOI: 10.1002/jcp.25876
       
  • Cover Image, Volume 232, Number 6, June 2017
    • Authors: María Laura Matos; Lara Lapyckyj, Marina Rosso, María José Besso, María Victoria Mencucci, Clara Isabel Marín Briggiler, Silvina Giustina, Laura Inés Furlong, Mónica Hebe Vazquez-Levin
      Abstract: Cover: The cover image, by Mónica Hebe Vazquez-Levin et al., is based on the Original Research Article Identification of a Novel Human E-Cadherin Splice Variant and Assessment of Its Effects Upon EMT-Related Events,
      DOI : 10.1002/jcp.25622.
      PubDate: 2017-02-24T12:14:04.686251-05:
       
  • Differential Expression of Estrogen Receptor Variants in Response to
           Inflammation Signals in Human Airway Smooth Muscle
    • Authors: Bharathi Aravamudan; Katelyn J. Goorhouse, Ghanashyam Unnikrishnan, Michael A. Thompson, Christina M. Pabelick, John R. Hawse, Y. S. Prakash, Venkatachalem Sathish
      Abstract: The prevalence of asthma is higher in pre-pubescent and aging males, and in post-pubertal females, strongly indicating that sex steroids (especially estrogen) may be an important modulator in lung disease. We recently demonstrated that airway smooth muscle (ASM) expresses both alpha and beta forms of the estrogen receptor (ERα and ERβ) in males and females, and that these receptors regulate intracellular [Ca2+] and ASM contractility. Although both ERα and ERβ have multiple splice variants, it is unclear if and how the expression of these variants is modulated under conditions such as chronic inflammation/asthma. In order to test the hypothesis that the differential expression of ERα and ERβ variants contributes to the pathogenesis of asthma, we profiled the expression of various ERα and ERβ genes in asthmatic and inflamed (TNFα- or IL-13-treated) ASM. Gene expression was assessed at both the mRNA and protein levels in asthmatic ASM cells or non-asthmatic cells treated with TNFα (20 ng/ml) or IL-13 (50 ng/ml). We observed marked variation in the expression of ER isoforms in response to inflammatory stimuli, and in non-asthmatic versus asthmatic ASM. Changes in protein levels of ERα and ERβ corresponded with the observed differential mRNA patterns. Pharmacological studies implicate cytosolic (p42/44 MAPK and PI3 K) and nuclear (NFκB, STAT6, and AP-1) signaling pathways as putative mechanisms that mediate and/or regulate effects of inflammation on ER expression. We conclude that variations in ASM ER expression profiles occur with inflammation and that ER variants could contribute to estrogen signaling in airway diseases such as asthma. J. Cell. Physiol. 9999: 1–7, 2016. © 2016 Wiley Periodicals, Inc.
      PubDate: 2017-02-24T09:10:37.512884-05:
      DOI: 10.1002/jcp.25674
       
  • T11TS Repress Gliomagenic Apoptosis of Bone Marrow Hematopoietic Stem
           Cells
    • Authors: Somnath Mondal; Iman Hazra, Ankur Datta, Omar Faruk Sk Md, Saibal Moitra, Santanu Kumar Tripathi, Swapna Chaudhuri
      Abstract: Combating gliomagenic global immunosuppression is the one of the emerging key for improving prognosis in malignant glioma. Apoptosis plays a pivotal role within the adult hematopoietic system particularly in regulating the cells of immune system. Gliomagenic regulation of apoptotic mediators within bone marrow milieu has not been elucidated. We previously demonstrated that administration of membrane glycopeptides T11 target structure (T11TS) not only rejuvenate bone marrow hematopoietic stem cells (BMHSCs) from glioma mediated hibernation by inhibiting gliomagenic overexpression of Ang-1/Tie-2 but also stimulate glioma mediated diminution of expression CD34, c-kit and Sca-1 markers. In the present study we investigated the impact of glioma on apoptotic signaling cascades of BMHSCs and consequences following T11TS therapy. Bone marrow smear and Annexin V staining confirm gliomagenic acceleration of apoptotic fate of BMHSCs whereas T11TS treatment in glioma-bearing rats disrupted apoptosis of BMHSCs. Flowcytometry, immunoblotting and immunofluorescence imagining results revealed multi potent T11TS not only significantly downregulates gliomagenic overexpression of Fas, Fas L, Bid and caspase-8, the pro-apoptotic extrinsic mediators but also strongly inhibits cytosolic release of cytochrome-c, Apf-1, and Bax to deactivate gliomagenic caspase-9, 3 the key intrinsic apoptotic mediators followed by up modulation of anti-apoptotic Bcl-2 in glioma associated HSCs. T11TS is also able to diminish the perforin-granzyme B mediated apoptotic verdict of BMHSCs during gliomagenesis. The anti-apoptotic action of T11TS on glioma associated BMHSCs provide a crucial insight into how T11TS exerts its immunomodulatory action against glioma mediated immune devastation. This article is protected by copyright. All rights reserved
      PubDate: 2017-02-24T04:40:25.484671-05:
      DOI: 10.1002/jcp.25874
       
  • Profilin1 Is Expressed in Osteocytes and Regulates Cell Shape and
           Migration
    • Authors: Wanting Lin; Yayoi Izu, Arayal Smriti, Makiri Kawasaki, Chantida Pawaputanon, Ralph T. Böttcher, Mercedes Costell, Keiji Moriyama, Masaki Noda, Yoichi Ezura
      Abstract: Osteocytes are the most abundant cells in bone and regulate bone metabolism in coordination with osteoblasts and osteoclasts. However, the molecules that control osteocytes are still incompletely understood. Profilin1 is an actin-binding protein that is involved in actin polymerization. Osteocytes possess characteristic dendritic process formed based on actin cytoskeleton. Here, we examined the expression of profilin1 and its function in osteocytes. Profilin1 mRNA was expressed in osteocytic MLO-Y4 cells and its levels were gradually increased along with the time in culture. With regard to functional aspect, knockdown of profilin1 by siRNA enhanced BMP-induced increase in alkaline phosphatase expression levels in MLO-Y4 cells. Profilin1 knockdown suppressed the levels of dendritic processes and migration of MLO-Y4 cells. Since ageing causes an increase in ROS in the body, we further examined the effects of hydrogen peroxide on the expression of profilin1. Hydrogen peroxide treatment increased the levels of profilin1 mRNA in MLO-Y4 cells in contrast to the decline in alkaline phosphatase. Profilin1 was expressed not only in MLO-Y4cells but also in the primary cultures of osteocytes. Importantly, profilin1 mRNA levels in primary cultures of osteocytes were higher than those in primary cultures of osteoblasts. To examine in vivo role of profilin1 in osteocytes, proflin1 was conditionally knocked out by using DMP1-cre and profilin1 floxed mice. This conditional deletion of profilin1 specifically in osteocytes resulted in reduction in the levels of bone volume and bone mineral density. These data indicate that profilin1 is expressed in osteocytes and regulates cell shape, migration and bone mass. This article is protected by copyright. All rights reserved
      PubDate: 2017-02-24T04:30:35.728431-05:
      DOI: 10.1002/jcp.25872
       
  • Soft Tissue Engineering with Micronized-Gingival Connective Tissues
    • Authors: Sawako Noda; Yoshinori Sumita, Seigo Ohba, Hideyuki Yamamoto, Izumi Asahina
      Abstract: The free gingival graft (FGG) and connective tissue graft (CTG) are currently considered to be the gold standards for keratinized gingival tissue reconstruction and augmentation. However, these procedures have some disadvantages in harvesting large grafts, such as donor-site morbidity as well as insufficient gingival width and thickness at the recipient site post-treatment. To solve these problems, we focused on an alternative strategy using micronized tissue transplantation (micro-graft). In this study, we first investigated whether transplantation of micronized gingival connective tissues (MGCTs) promotes skin wound healing. MGCTs (≤100 µm) were obtained by mincing a small piece (8 mm3) of porcine keratinized gingiva using the RIGENERA system. The MGCTs were then transplanted to a full skin defect (5 mm in diameter) on the dorsal surface of immunodeficient mice after seeding to an atelocollagen matrix. Transplantations of atelocollagen matrixes with and without micronized dermis were employed as experimental controls. The results indicated that MGCTs markedly promote the vascularization and epithelialization of the defect area 14 days after transplantation compared to the experimental controls. After 21 days, complete wound closure with low contraction was obtained only in the MGCT grafts. Tracking analysis of transplanted MGCTs revealed that some mesenchymal cells derived from MGCTs can survive during healing and may function to assist in wound healing. We propose here that micro-grafting with MGCTs represents an alternative strategy for keratinized tissue reconstruction that is characterized by low morbidity and ready availability. This article is protected by copyright. All rights reserved
      PubDate: 2017-02-24T04:30:32.232091-05:
      DOI: 10.1002/jcp.25871
       
  • MicroRNA106a Regulates Matrix Metalloprotease 9 in a Sirtuin Dependent
           Mechanism
    • Authors: Lincy Edatt; Ashutosh Kumar Maurya, Grace Raji, Haritha K, Sameer V.B. Kumar
      Abstract: Cellular migration is important during many physiological as well as pathological conditions and is regulated very tightly by an intricate network of signaling and effecter molecules. One of the important players during cellular migration are matrix metalloproteases and their levels have been reported to be important in determining the cellular migratory properties during metastasis. MMPs and regulators of MMPs therefore, present themselves as potent candidates for manipulation, to control conditions where they get dysregulated. Micro RNAs are a group of micro regulators that can modulate expression of a gene through transcriptional and post transcriptional regulations. Owing to the fact that many microRNAs have already been reported to regulate MMPs and that miR106a, a member of oncomir17 family has been implicated in metastatic conditions, the present study intended to analyze if miR106a can regulate levels of MMP9, an important inducible matrix metalloproteinase. The results of thein vitro experiments demonstrated that under conditions of migration cells showed elevated levels of miR106a, which could regulate the expression of major MMP9 regulator, SIRT-1. Decreased levels of SIRT1thus resulted in an increase in the expression and activity of MMP9. Over expression and mRNA stability studies carried out also suggested regulatory role of miR106a. The overall results thus suggested that the levels of miR106a gets modulated during cellular migration, causing a change in the levels of SIRT-1 mRNA by affecting its stability and the levels of SIRT-1 in turn can regulate the levels of MMP9. This article is protected by copyright. All rights reserved
      PubDate: 2017-02-24T04:30:28.139507-05:
      DOI: 10.1002/jcp.25870
       
  • Adenosine and Adenosine Receptors in the Immunopathogenesis and Treatment
           of Cancer
    • Authors: Mohammad Hossein Kazemi; Sahar Raoufi, Mohammad Hojjat-Farsangi, Enayat Anvari, Ghasem Ghalamfarsa, Hamed Mohammadi, Farhad Jadidi-Niaragh
      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 including A1, A2A, A2B, and A3 which are expressed on various immune cells. Several studies have indicated the overexpression of adenosine generating enzymes and adenosine receptors in various cancers which was correlated with tumor progression. Since the signaling of adenosine receptors enhances tumor progression, their manipulation can be promising therapeutic approach in cancer therapy. Accordingly, several agonists and antagonists against adenosine receptors have been designed for cancer therapy. In this review, we will try to clarify the role of different adenosine receptors in the immunopathogenesis, as well as their role in the treatment of cancer. This article is protected by copyright. All rights reserved
      PubDate: 2017-02-24T04:30:26.555417-05:
      DOI: 10.1002/jcp.25873
       
  • Cytoplasmic Localization of RUNX3 via Histone Deacetylase-Mediated SRC
           Expression in Oxidative-Stressed Colon Cancer Cells
    • Authors: Kyoung Ah Kang; Mei Jing Piao, Yea Seong Ryu, Young Hee Maeng, Jin Won Hyun
      Abstract: Runt domain transcription factor 3 (RUNX3) is a transcription factor that functions as a tumor suppressor. RUNX3 is frequently inactivated by epigenetic silencing or its protein mislocalization (cytoplasmic localization) in many cancer types. This study investigated whether oxidative stress induces redistribution of RUNX3 from the nucleus to the cytoplasm. The cytoplasmic localization of RUNX3 was associated with oxidative stress-induced RUNX3 phosphorylation at tyrosine residues via SRC activation. Moreover, oxidative stress increased expression of histone deacetylases (HDACs). RUNX3 phosphorylation and SRC expression induced by oxidative stress were inhibited by knockdown of HDAC1, restoring the nuclear localization of RUNX3 under oxidative stress. In conclusion, these results demonstrate that HDAC1- and SRC-mediated phosphorylation of RUNX3 induced by oxidative stress is associated with the cytoplasmic localization of RUNX3 and can lead to RUNX3 inactivation and carcinogenesis. J. Cell. Physiol. 9999: 1–8, 2016. © 2016 Wiley Periodicals, Inc.RUNX3 cytoplasmic localization induced by oxidative stress in colon cancer cells. Oxidative stress induces HDAC1 expression and then activates SRC. Activated SRC induces expression of phospho-RUNX3. Phospho-RUNX3 is retained in the cytoplasm, leading to degradation of cytoplasmic RUNX3 by the proteasome system and finally resulting carcinogenesis.
      PubDate: 2017-02-21T05:31:21.597574-05:
      DOI: 10.1002/jcp.25746
       
  • STIM-1 and ORAI-1 channel mediate angiotensin-II-induced expression of
           Egr-1 in vascular smooth muscle cells
    • Authors: Estelle R. Simo-Cheyou; Ju Jing Tan, Ryszard Grygorczyk, Ashok K. Srivastava
      Abstract: An upregulation of Egr-1 expression has been reported in models of atherosclerosis and intimal hyperplasia and, various vasoactive peptides and growth promoting stimuli have been shown to induce the expression of Egr-1 in vascular smooth muscle cells (VSMC). Angiotensin-II (Ang-II) is a key vasoactive peptide that has been implicated in the pathogenesis of vascular diseases. Ang-II elevates intracellular Ca2+ through activation of the store-operated calcium entry (SOCE) involving an inositol-3-phosphate receptor (IP3R)-coupled depletion of endoplasmic reticular Ca2+ and a subsequent activation of the stromal interaction molecule 1 (STIM-1)/Orai-1 complex. However, the involvement of IP3R/STIM-1/Orai-1-Ca2+-dependent signaling in Egr-1 expression in VSMC remains unexplored. Therefore, in the present studies, we have examined the role of Ca2+ signaling in Ang-II-induced Egr-1 expression in VSMC and investigated the contribution of STIM-1 or Orai-1 in mediating this response. 2-aminoethoxydiphenyl borate (2-APB), a dual non-competitive antagonist of IP3R and inhibitor of SOCE, decreased Ang-II-induced Ca2+ release and attenuated Ang-II-induced enhanced expression of Egr-1 protein and mRNA levels. Egr-1 upregulation was also suppressed following blockade of calmodulin and CaMKII. Furthermore, RNA interference-mediated depletion of STIM-1 or Orai-1 attenuated Ang-II-induced Egr-1 expression as well as Ang-II-induced phosphorylation of ERK1/2 and CREB. In addition, siRNA-induced silencing of CREB resulted in a reduction in the expression of Egr-1 stimulated by Ang-II. In summary, our data demonstrate that Ang-II-induced Egr-1 expression is mediated by STIM-1/Orai-1/Ca2+-dependent signaling pathways in A-10 VSMC.Angiotensin-II-induced intracellular calcium release depletes the endoplasmic reticulum store, resulting in the activation of the stromal interaction molecule 1 (STIM1) known to cooperate with Orai-1 transmembrane channel in mediating store-operated calcium entry (SOCE) in the cells. In this study, we demonstrate that intact SOCE and unaltered expression of STIM-1 and Orai-1, in association with an involvement of calcium signaling molecules such as calmodulin and CaMKII, are required for Ang-II-dependent induction of the early growth response protein-1 in VSMC. We also provide evidence of the critical role played by ERK1/2 and CREB activation in mediating this response.
      PubDate: 2017-02-21T05:21:00.969387-05:
      DOI: 10.1002/jcp.25810
       
  • Regulation of mitotic spindle orientation during epidermal stratification
    • Authors: Wei Xie; Jun Zhou
      Abstract: The epidermis is a stratified epithelium that serves as a barrier to infection from environmental pathogens and prevents water loss. Epidermal stratification is tightly controlled during embryogenesis. Progenitor cells in the developing epidermis undergo both symmetric and asymmetric cell divisions to balance the growth of the skin surface area against the generation of differentiated cell layers. Therefore, understanding the relationship between oriented divisions of progenitor cells and the development and stratification of the epidermis is of paramount importance in the field of skin biology and pathology. We provide here an integrated view of recent studies implicating that improper orientation of the mitotic spindle contributes to disorders associated with abnormal epidermal stratification and suggesting that spindle orientation could serve as a potential therapeutic target in skin diseases.We provide here an integrated view of recent studies implicating that improper orientation of the mitotic spindle contributes to disorders associated with abnormal epidermal stratification. We also suggest that spindle orientation could serve as a potential therapeutic target in skin diseases.
      PubDate: 2017-02-21T05:15:49.12907-05:0
      DOI: 10.1002/jcp.25750
       
  • LCN2 overexpression in bone enhances the hematopoietic compartment via
           modulation of the bone marrow microenvironment
    • Authors: Delfina Costa; Elisa Principi, Edoardo Lazzarini, Fiorella Descalzi, Ranieri Cancedda, Patrizio Castagnola, Sara Tavella
      Abstract: Lipocalin-2 (LCN2) is a member of the lipocalin family whose expression is modulated in several conditions, including cell differentiation, innate immunity, stress, and cancer. Although it is known that it is expressed in bone, its function in this tissue remains poorly studied. To this end, we took advantage of transgenic mice lines that expressed LCN2 driven by a bone specific type I collagen (LCN2-Tg). In the bone marrow (BM) of LCN2-Tg mice we observed an increased number of phenotypically long-term hematopoietic stem cells (LT-HSC) that also displayed a higher proliferation rate compared to wild-type controls (Wt). Furthermore, hematopoietic progenitor cells, obtained from LCN2-Tg BM showed an increased clonogenic capacity compared to those obtained from LCN2-Tg spleen, a higher concentration of serum erythropoietin and a higher number of mature erythrocytes in the peripheral blood of old LCN2-Tg animals compared to aged-matched wt. The findings of a combined increase in the BM of the LCN2-Tg mice of SDF-1, SCF, and TIMP-1 levels along with the reduction of both MMP-9 activity and cathepsin K concentration may explain the observed effects on the HSC compartment. This study shows that LCN2 overexpression in bones modifies the BM microenvironment via modulation of the expression of key secreted factors and cytokines, which in turn regulate the HSC niche behavior enhancing both HSC homing in young mice and erythrocytes production in older mice.The overexpression of LCN2 in bone stimulates the expansion of the HSC by inducing pro-retention factors and inhibiting pro-egress signals for hematopoietic stem cells.
      PubDate: 2017-02-21T05:10:50.678787-05:
      DOI: 10.1002/jcp.25755
       
  • Na+/H+ exchanger NHE1 and NHE2 have opposite effects on migration velocity
           in rat gastric surface cells
    • Authors: Anja Paehler vor der Nolte; Giriprakash Chodisetti, Zhenglin Yuan, Florian Busch, Brigitte Riederer, Min Luo, Yan Yu, Manoj B. Menon, Andreas Schneider, Renata Stripecke, Katerina Nikolovska, Sunil Yeruva, Ursula Seidler
      Abstract: Following superficial injury, neighbouring gastric epithelial cells close the wound by rapid cell migration, a process called epithelial restitution. Na+/H+ exchange (NHE) inhibitors interfere with restitution, but the role of the different NHE isoforms expressed in gastric pit cells has remained elusive. The role of the basolaterally expressed NHE1 (Slc9a1) and the presumably apically expressed NHE2 (Slc9a2) in epithelial restitution was investigated in the nontransformed rat gastric surface cell line RGM1. Migration velocity was assessed by loading the cells with the fluorescent dye DiR and following closure of an experimental wound over time. Since RGM1 cells expressed very low NHE2 mRNA and have low transport activity, NHE2 was introduced by lentiviral gene transfer. In medium with pH 7.4, RGM1 cells displayed slow wound healing even in the absence of growth factors and independently of NHE activity. Growth factors accelerated wound healing in a partly NHE1-dependent fashion. Preincubation with acidic pH 7.1 stimulated restitution in a NHE1-dependent fashion. When pH 7.1 was maintained during the restitution period, migratory speed was reduced to ∼10% of the speed at pH 7,4, and the residual restitution was further inhibited by NHE1 inhibition. Lentiviral NHE2 expression increased the steady-state pHi and reduced the restitution velocity after low pH preincubation, which was reversible by pharmacological NHE2 inhibition. The results demonstrate that in RGM1 cells, migratory velocity is increased by NHE1 activation, while NHE2 activity inhibit this process. A differential activation of NHE1 and NHE2 may therefore, play a role in the initiation and completion of the epithelial restitution process.Genetic deletion of the Na+/H+ exchanger NHE2 leads to compromised wound healing in the gastrointestinal tract of mice. We studied the roles of NHE1 and NHE2 in gastrointestinal wound healing in vitro, and while NHE1 activity supported wound healing velocity, NHE2/GFP-overexpression dramatically reduced wound healing velocity compared to GFP expression alone. This effect was abolished by a NHE2- but not a NHE1 inhibitory concentration of HOE642. Thus, NHE1 and NHE2 may be important for the initiation and the resolution of gastrointestinal wound healing.
      PubDate: 2017-02-21T05:05:58.470701-05:
      DOI: 10.1002/jcp.25758
       
  • Osteogenic differentiation of human gingival mesenchymal stem cells by
           Aristolochia bracteolata supplementation through enhanced Runx2 expression
           
    • Authors: Dinesh Murugan Girija; Suresh Y Ranga Rao, Mangathayaru Kalachaveedu, Rajasekaran Subbarayan
      Abstract: The phenotypic characteristics of human gingival derived mesenchymal stem cells (HGMSCs) on induction with total methanol extract of Aristolochia bracteolata have been evaluated. HGMSCs were cultured in control and two different induction medium: Control medium (basal medium), OM1 (Standard induction medium), and OM2 (100 μg/ml of A. bracteolata). Osteogenic differentiation of the cultured cells was assessed by studying the calcium deposition and osteoblastic gene expression. OM2 medium showed an enhanced osteogenic differentiation potential than OM1 as measured by increased calcium deposition and elevated expression of Runx2, osteopontin, osteonectin, osteocalcin, Collagen type I, and ALP levels in comparison with OM1 differentiated cells. We conclude that at 100 μg/ml A. bracteolata has induced HGMSC differentiation into osteogenic lineage consequent to enhanced Runx2 expression and related osteogenic genes.The phenotypic characteristics of human gingival derived mesenchymal stem cells (HGMSCs) on induction with total methanol extract of Aristolochia bracteolata have been evaluated. A. bracteolata has induced HGMSC differentiation into osteogenic lineage consequent to enhanced Runx2 expression and related osteogenic genes.
      PubDate: 2017-02-21T04:55:23.652165-05:
      DOI: 10.1002/jcp.25835
       
  • Long noncoding RNAs (lncRNAs) in triple negative breast cancer
    • Authors: Qiuhong Wang; Sheng Gao, Haibo Li, Mingming Lv, Cheng Lu
      Abstract: Long noncoding RNAs (lncRNAs) are dysregulated in many cancer types, which are believed to play crucial roles in regulating several hallmarks of cancer biology. Triple Negative Breast Cancer (TNBC) is a very aggressive subtype of normal breast cancer, which has features of negativity for ER, PR, and HER2. Great efforts have been made to identify an association between lncRNAs expression profiles and TNBC, and to understand the functional role and molecular mechanism on aberrant-expressed lncRNAs. In this review, we summarized the existed knowledge on the systematics, biology, and function of lncRNAs. The advances from the most recent studies of lncRNAs in the predicament of breast cancer, TNBC, are highlighted, especially the functions of specifically selected lncRNAs. We also discussed the potential value of these lncRNAs in TNBC, providing clues for the diagnosis and treatments of TNBC.
      PubDate: 2017-02-21T04:50:25.364351-05:
      DOI: 10.1002/jcp.25830
       
  • Activin-SMAD signaling is required for maintenance of porcine iPS cell
           self-renewal through upregulation of NANOG and OCT4 expression
    • Authors: Fan Yang; Ning Wang, Yaxian Wang, Tong Yu, Huayan Wang
      Abstract: Porcine induced pluripotent stem cells (piPSCs) retain the enormous potential for farm animal reproduction and translational medicine, and have been reported by many laboratories worldwide. Some piPSC lines were bFGF-dependence and showed mouse EpiSC-like morphology; other lines were LIF-dependence and showed mouse ESC-like morphology. Metastable state of piPSC line that required both LIF and bFGF was also reported. Because bona fide pig embryonic stem cells were not available, uncovering piPSC state-specific regulatory circuitries was the most important task. In this study, we explored the function of Activin-SMAD signaling pathway and its downstream activated target genes in piPSCs. Transcriptome analysis showed that genes involved in Activin-SMAD signaling pathway were evidently activated during porcine somatic cell reprogramming, regardless piPSCs were LIF- or bFGF-dependent. Addition of Activin A and overexpression of SMAD2/3 significantly promoted expressions of porcine NANOG and OCT4, whereas inhibition of Activin-SMAD signaling by SB431542 and SMAD7 reduced NANOG and OCT4 expressions, and induced piPSCs differentiation exiting from pluripotent state. Our data demonstrate that activation of Activin-SMAD signaling pathway by addition of Activin A in culture medium is necessary for maintenance of self-renewal in porcine pluripotent stem cells.Our data demonstrate that activation of Activin-SMAD signaling pathway by addition of Activin A in culture medium is necessary for maintenance of self-renewal in porcine pluripotent stem cells.
      PubDate: 2017-02-21T04:40:33.007513-05:
      DOI: 10.1002/jcp.25747
       
  • Glucose Adsorption to Chitosan Membranes Increases Proliferation of Human
           Chondrocyte via Mammalian Target of Rapamycin Complex 1 and Sterol
           Regulatory Element-binding Protein-1 Signaling
    • Authors: Shun-Fu Chang; Kuo-Chin Huang, Chin-Chang Cheng, Yu-Ping Su, Ko-Chao Lee, Cheng-Nan Chen, Hsin-I Chang
      Abstract: BackgroundOsteoarthritis (OA) is currently still an irreversible degenerative disease of the articular cartilage. Recent, dextrose (D-glucose) intraarticular injection prolotherapy for OA patients has been reported to benefit the chondrogenic stimulation of damaged cartilage. However, the detailed mechanism of glucose's effect on cartilage repair remains unclear. Chitosan, a naturally derived polysaccharide, has recently been investigated as a surgical or dental dressing to control breeding. Therefore, in this study, glucose was adsorbed to chitosan membranes (CTS-Glc), and the study aimed to investigate whether CTS-Glc complex membranes could regulate the proliferation of human OA chondrocytes and to explore the underlying mechanism.MethodsHuman OA and SW1353 chondrocytes were used in this study. The experiments involving the transfection of cells used SW1353 chondrocytes. A specific inhibitor and siRNAs were used to investigate the mechanism underlying the CTS-Glc-regulated proliferation of human chondrocytes.ResultsWe found that CTS-Glc significantly increased the proliferation of both human OA and SW1353 chondrocytes comparable to glucose- or chitosan-only stimulation. The role of mammalian target of rapamycin complex 1 (mTORC1) signaling, including mTOR, raptor, and S6k proteins, has been demonstrated in the regulation of CTS-Glc-increased human chondrocyte proliferation. mTORC1 signaling increased the expression levels of maturated SREBP-1 and FASN and then induced the expressions of cell cycle regulators, i.e., cyclin D, cyclin-dependent kinase-4 and -6 in human chondrocytes.ConclusionsThis study elucidates the detailed mechanism behind the effect of CTS-Glc complex membranes in promoting chondrocyte proliferation and proposes a possible clinical application of the CTS-Glc complex in the dextrose intraarticular injection of OA prolotherapy in the future to attenuate the pain and discomfort of OA patients. This article is protected by copyright. All rights reserved
      PubDate: 2017-02-20T07:51:23.531003-05:
      DOI: 10.1002/jcp.25869
       
  • MAPK3/1 Participates in the Activation of Primordial Follicles through
           mTORC1-KITL Signaling
    • Authors: Yu Zhao; Yu Zhang, Jia Li, Nana Zheng, Xiaoting Xu, Jing Yang, Guoliang Xia, Meijia Zhang
      Abstract: The majority of ovarian primordial follicles are preserved in a dormant state to maintain the female reproductive lifespan, and only a few primordial follicles are activated to enter the growing follicle pool in each wave. Recent studies have shown that primordial follicular activation depends on mammalian target of rapamycin complex 1 (mTORC1)-KIT ligand (KITL) signaling in pre-granulosa cells and its receptor (KIT)-phosphoinositol 3 kinase (PI3K) signaling in oocytes. However, the upstream regulator of mTORC1 signaling is unclear. The results of the present study showed that the phosphorylated mitogen-activated protein kinase3/1 (MAPK3/1) protein is expressed in some primordial follicles and all growing follicles. Culture of 3 days post-parturition (dpp) ovaries with the MAPK3/1 signaling inhibitor U0126 significantly reduced the number of activated follicles and was accompanied by dramatically reduced granulosa cell proliferation and increased oocyte apoptosis. Western blot and immunofluorescence analyses showed that U0126 significantly decreased the phosphorylation levels of Tsc2, S6K1, and rpS6 and the expression of KITL, indicating that U0126 inhibits mTORC1-KITL signaling. Furthermore, U0126 decreased the phosphorylation levels of Akt, resulting in a decreased number of oocytes with Foxo3 nuclear export. To further investigate MAPK3/1 signaling in primordial follicle activation, we used phosphatase and tensin homolog deleted on chromosome 10 (PTEN) inhibitor bpV(HOpic) to promote primordial follicle activation. In this model, U0126 also inhibited the activation of primordial follicles and mTORC1 signaling. Thus, these results suggest that MAPK3/1 participates in primordial follicle activation through mTORC1-KITL signaling. This article is protected by copyright. All rights reserved
      PubDate: 2017-02-20T07:45:26.212931-05:
      DOI: 10.1002/jcp.25868
       
  • Palmitate Induces Mitochondrial Superoxide Generation and Activates AMPK
           in Podocytes
    • Authors: Eugene Lee; Jin Choi, Hyun Soon Lee
      Abstract: Studies have shown that high levels of serum free fatty acids (FFAs) are associated with lipotoxicity and type 2 diabetes. Palmitic acid (PA) is the predominant circulating saturated FFA, yet its role in the pathogenesis of diabetic nephropathy (DN) is not clear. Recently, one study suggested that mitochondrial superoxide production is related to AMP-activated protein kinase (AMPK) activity in diabetic mice kidneys. To elucidate the link between PA and oxidative stress and AMPK activity in DN, we compared the cultured murine podocytes exposed to PA and oleic acid (OA). Incubation of cells with 250 µM PA or OA induced a translocation of CD36, a fatty acid transport protein, with intracellular lipid accumulation. PA, but not OA, induced mitochondrial superoxide and hydrogen peroxide (H2O2) generation in podocytes, as shown by enhanced fluorescence of MitoSOX Red and dichlorofluorescein (DCF), respectively. Costimulation of PA-treated cells with the H2O2 scavenger catalase abolished the PA-induced DCF fluorescence. Only PA induced mitochondrial damage as shown by electron microscopy. The AMPK activity was determined by immunobloting, measuring the ratio of phosphorylated AMPK (p-AMPK) to total AMPK. Only PA significantly increased the p-AMPK levels compared with controls. Addition of catalase to PA-treated cells did not affect the PA-stimulated p-AMPK levels. Collectively, our results indicate that PA induces mitochondrial superoxide and H2O2 generation in cultured podocytes, which may not be directly linked to AMPK activation. Given that, PA seems to play an important role in the pathogenesis of DN through lipotoxicity initiated by mitochondrial superoxide overproduction. This article is protected by copyright. All rights reserved
      PubDate: 2017-02-18T10:00:27.672401-05:
      DOI: 10.1002/jcp.25867
       
  • Mitochondria-targeted Esculetin Inhibits PAI-1 Levels by Modulating STAT3
           Activation and miR-19b via SIRT3: Role in Acute Coronary Artery Syndrome
    • Authors: Sujana Katta; Santosh Karnewar, Devayani Panuganti, Mahesh Kumar Jerald, BK Sastry, Srigiridhar Kotamraju
      Abstract: In this study, we explored the microRNAs responsible for the regulation of PAI-1 during LPS-stimulated inflammation in human aortic endothelial cells, subsequently studied the effect of a newly synthesized mitochondria-targeted esculetin (Mito-Esc) that was shown for its anti-atherosclerotic potential in modulating PAI-1 levels and its targeted miRs during angiotensin-II-induced atherosclerosis in ApoE-/- mice. LPS-stimulated PAI-1 was accompanied with an upregulation of miR-19b and down-regulation of miR-30c. These effects of LPS on PAI-1 were reversed in the presence of both parent esculetin and Mito-Esc. However, the effect of Mito-Esc was more pronounced in the regulation of PAI-1. In addition, LPS-stimulated PAI-1 expression was significantly decreased in cells treated with Anti-miR-19b, thereby suggesting that miR-19b co-expression plays a key role in PAI-1 regulation. The results also show that incubation of cells with Stattic, an inhibitor of STAT-3, inhibited LPS-stimulated PAI-1 expression. Interestingly, knockdown of SIRT3, a mitochondrial biogenetic marker, enhanced PAI-1 levels via modulation of miR-19b and -30c. Mito-Esc treatment significantly inhibited Ang-II-induced PAI-1, possibly via altering miR-19b and 30c in ApoE-/- mice. The association between PAI-1, miR-19b and -30c were further confirmed in plasma and microparticles isolated from patients suffering from acute coronary syndrome of various degrees. Taken together, LPS-induced PAI-1 involves co-expression of miR-19b and down regulation of miR-30c and Mito-Esc treatment by modulating miR-19b and miR-30c through SIRT3 activation, inhibits PAI-1 levels that, in part, contribute to its anti-atherosclerotic effects. Moreover, there exists a strong positive correlation between miR-19b and PAI-1 in patients suffering from ST-elevated myocardial infarction. This article is protected by copyright. All rights reserved
      PubDate: 2017-02-18T05:20:37.618848-05:
      DOI: 10.1002/jcp.25865
       
  • Transforming Growth Factor β Induces Bone Marrow Mesenchymal Stem Cell
           Migration via Noncanonical Signals and N-cadherin
    • Authors: Maria Jose Dubon; Jinyeong Yu, Sanghyuk Choi, Ki-Sook Park
      Abstract: Transforming growth factor-beta (TGF-β) induces the migration and mobilization of bone marrow-derived mesenchymal stem cells (BM-MSCs) to maintain bone homeostasis during bone remodeling and facilitate the repair of peripheral tissues. Although many studies have reported the mechanisms through which TGF-β mediates the migration of various types of cells, including cancer cells, the intrinsic cellular mechanisms underlying cellular migration and mobilization of BM-MSCs mediated by TGF-β are unclear. In this study, we showed that TGF-β activated noncanonical signaling molecules, such as Akt, extracellular signal-regulated kinase 1/2 (ERK1/2), focal adhesion kinase (FAK), and p38, via TGF-β type I receptor in human BM-MSCs and murine BM-MSC-like ST2 cells. Inhibition of Rac1 by NSC23766 and Src by PP2 resulted in impaired TGF-β-mediated migration. These results suggested that the Smad-independent, noncanonical signals activated by TGF-β were necessary for migration. We also showed that N-cadherin-dependent intercellular interactions were required for TGF-β-mediated migration using functional inhibition of N-cadherin with EDTA treatment and a neutralizing antibody (GC-4 antibody) or siRNA-mediated knockdown of N-cadherin. However, N-cadherin knockdown did not affect the global activation of noncanonical signals in response to TGF-β. Therefore, these results suggested that the migration of BM-MSCs in response to TGF-β was mediated through N-cadherin and noncanonical TGF-β signals. This article is protected by copyright. All rights reserved
      PubDate: 2017-02-18T05:20:33.723598-05:
      DOI: 10.1002/jcp.25863
       
  • Subclinical alteration of the cervical–vaginal microbiome in women
           with idiopathic infertility
    • Authors: Giuseppina Campisciano; Fiorella Florian, Angela D'Eustacchio, David Stanković, Giuseppe Ricci, Francesco De Seta, Manola Comar
      Abstract: Biomarkers have a wide application in research and clinic, they help to choose the correct treatment for diseases. Recent studies, addressing the vaginal microbiome using next generation sequencing (NGS), reported the involvement of bacterial species in infertility. We compared the vaginal microbiome of idiopathic infertile women with that of healthy, including bacterial vaginosis affected women and non-idiopathic infertile women, to identify bacterial species suitable as biomarkers. Information on microorganisms was obtained from the V3-16S rDNA sequencing of cervical–vaginal fluids of 96 women using the Ion Torrent platform. Data were processed with QIIME and classified against the Vaginal 16S rDNA Reference Database. The analysis revealed a significant beta-diversity variation (p 
      PubDate: 2017-02-16T10:35:27.562047-05:
      DOI: 10.1002/jcp.25806
       
  • Mesenchyme Homeobox 2 Enhances Migration of Endothelial Colony Forming
           Cells Exposed to Intrauterine Diabetes Mellitus
    • Authors: Cassandra R. Gohn; Emily K. Blue, BreAnn M. Sheehan, Kaela M. Varberg, Laura S. Haneline
      Abstract: Diabetes mellitus (DM) during pregnancy has long-lasting implications for the fetus, including cardiovascular morbidity. Previously, we showed that endothelial colony forming cells (ECFCs) from DM human pregnancies have decreased vasculogenic potential. Here, we evaluate whether the molecular mechanism responsible for this phenotype involves the transcription factor, Mesenchyme Homeobox 2 (MEOX2). In human umbilical vein endothelial cells, MEOX2 upregulates cyclin-dependent kinase inhibitor expression, resulting in increased senescence and decreased proliferation. We hypothesized that dysregulated MEOX2 expression in neonatal ECFCs from DM pregnancies decreases network formation through increased senescence and altered cell cycle progression. Our studies show that nuclear MEOX2 is increased in ECFCs from DM pregnancies. To determine if MEOX2 is sufficient and/or required to induce impaired network formation, MEOX2 was overexpressed and depleted in ECFCs from control and DM pregnancies, respectively. Surprisingly, MEOX2 overexpression in control ECFCs resulted in increased network formation, altered cell cycle progression, and increased senescence. In contrast, MEOX2 knockdown in ECFCs from DM pregnancies led to decreased network formation, while cell cycle progression and senescence were unaffected. Importantly, migration studies demonstrated that MEOX2 overexpression increased migration, while MEOX2 knockdown decreased migration. Taken together, these data suggest that altered migration may be mediating the impaired vasculogenesis of ECFCs from DM pregnancies. While initially believed to be maladaptive, these data suggest that MEOX2 may serve a protective role, enabling increased vessel formation despite exposure to a DM intrauterine environment. J. Cell. Physiol. 9999: 1–8, 2016. © 2016 Wiley Periodicals, Inc.Endothelial colony forming cells (ECFCs) from diabetic human pregnancies have decreased vasculogenic potential both in vitro and in vivo. The goal of this study was to evaluate whether the molecular mechanism responsible for this phenotype involves the transcription factor, Mesenchyme Homeobox 2 (MEOX2). While initially believed to be maladaptive, these data suggest that MEOX2 may serve a protective role, enabling increased vessel formation despite exposure to a DM intrauterine environment.
      PubDate: 2017-02-16T10:25:26.580862-05:
      DOI: 10.1002/jcp.25734
       
  • The Role of Cardiac Myosin Binding Protein C3 in Hypertrophic
           Cardiomyopathy-Progress and Novel Therapeutic Opportunities
    • Authors: Iman A. Mohamed; Navaneethakrishnan T. Krishnamoorthy, Gheyath K. Nasrallah, Sahar I. Da'as
      Abstract: Hypertrophic cardiomyopathy (HCM) is a common autosomal dominant genetic cardiovascular disorder marked by genetic and phenotypic heterogeneity. Mutations in the gene encodes the cardiac myosin-binding protein C, cMYBPC3 is amongst the various sarcomeric genes that are associated with HCM. These mutations produce mutated mRNAs and truncated cMyBP-C proteins. In this review, we will discuss the implications and molecular mechanisms involved in MYBPC3 different mutations. Further, we will highlight the novel targets that can be developed into potential therapeutics for the treatment of HMC. J. Cell. Physiol. 9999: 1–10, 2016. © 2016 Wiley Periodicals, Inc.This review discusses the implications and the and molecular mechanisms of MYBPC3 different mutations. Further, we will highlight the novel targets that can be developed into potential therapeutics for the treatment of Hypertrophic cardiomyopathy.
      PubDate: 2017-02-16T10:15:27.995734-05:
      DOI: 10.1002/jcp.25639
       
  • Identification of MSX1 and DCLK1 as mRNA Biomarkers for Colorectal Cancer
           Detection Through DNA Methylation Information
    • Authors: Ai-Jun Sun; Hai-Bo Gao, Gao Liu, Heng-Fa Ge, Zun-Ping Ke, Sen Li
      Abstract: Colorectal cancer is the second most deadly malignancy in the United States. However, the currently screening options had their limitation. Novel biomarkers for colorectal cancer detections are necessary to reduce the mortality. The clinical information, mRNA expression levels and DNA methylation information of colorectal cancer were downloaded from TCGA. The patients were separated into training group and testing group based on their platforms for DNA methylation. Beta values of DNA methylation from tumor tissues and normal tissues were utilized to figure out the position that were differentially methylated. The expression levels of mRNA of thirteen genes, whose CpG islands were differentially methylated, were extracted from the RNA-Seq results from TCGA. The probabilities whether the mRNA was differentially expressed between tumor and normal samples were calculated using Student's t-test. Logistic regression and decision tree were built for cancer detection and their performances were evaluated by the area under the curve (AUC). Twenty-four genomic locations were differentially methylated, which could be mapped to eleven genes. Nine out of eleven genes had differentially expressed mRNA levels, which were used to build the model for cancer detection. The final detection models consisting of mRNA expression levels of these nine genes had great performances on both training group and testing group. The model that constructed in this study suggested MSX1 and DCLK1 might be used in colorectal cancer detection or as target of cancer therapies. J. Cell. Physiol. 9999: 1–6, 2016. © 2016 Wiley Periodicals, Inc.Nine out of eleven genes had differentially expressed mRNA levels, which were used to build the model for cancer detection. The final detection models consisting of mRNA expression levels of these nine genes had great performances on both training group and testing group. The model that constructed in this study suggested MSX1 and DCLK1 might be used in colorectal cancer detection or as target of cancer therapies.
      PubDate: 2017-02-16T10:10:26.023949-05:
      DOI: 10.1002/jcp.25733
       
  • MicroRNA-125a-5p Is a Downstream Effector of Sorafenib in Its
           Antiproliferative Activity Toward Human Hepatocellular Carcinoma Cells
    • Authors: Nicoletta Potenza; Nicola Mosca, Silvia Zappavigna, Filomena Castiello, Marta Panella, Carmela Ferri, Daniela Vanacore, Antonio Giordano, Paola Stiuso, Michele Caraglia, Aniello Russo
      Abstract: Sorafenib is an antitumor drug for treatment of advanced hepatocellular carcinoma (HCC). It acts as a multikinase inhibitor suppressing cell proliferation and angiogenesis. Human microRNA-125a-5p (miR-125a) is endowed with similar activities and is frequently downregulated in HCC. Looking for a potential microRNA-based mechanism of action of the drug, we found that sorafenib increases cellular expression of miR-125a in cultured HuH-7 and HepG2 HCC cells. Upregulation of the microRNA inhibited cell proliferation by suppression of sirtuin-7, a NAD(+)-dependent deacetylase, and p21/p27-dependent cell cycle arrest in G1. Later, recruitment of miR-125a in the antiproliferative activity of sorafenib was inquired by modulating its expression in combination with the drug treatment. This analysis showed that intracellular delivery of miR-125a had no additive effect on the antiproliferative activity of sorafenib, whereas a miR-125a inhibitor could counteract it. Finally, evaluation of other oncogenic targets of miR-125a revealed its ability to interfere with the expression of matrix metalloproteinase-11, Zbtb7a proto-oncogene, and c-Raf, possibly contributing to the antiproliferative activity of the drug. J. Cell. Physiol. 9999: 1–7, 2016. © 2016 Wiley Periodicals, Inc.Upregulation of the microRNA inhibited cell proliferation by suppression of sirtuin-7, a NAD(+)-dependent deacetylase, and p21/p27-dependent cell cycle arrest in G1. Intracellular delivery of miR-125a had no additive effect on the antiproliferative activity of sorafenib, whereas a miR-125a inhibitor could counteract it. miR-125a interfered with the expression of matrix metalloproteinase-11, Zbtb7a proto-oncogene, and c-Raf, possibly contributing to the antiproliferative activity of the drug.
      PubDate: 2017-02-16T10:05:25.695604-05:
      DOI: 10.1002/jcp.25744
       
  • Dact1, a Wnt-Pathway Inhibitor, Mediates Human Mesangial Cell
           TGF-β1-Induced Apoptosis
    • Authors: Daniele Pereira Jardim; Paula Cristina Eiras Poço, Alexandre Holthausen Campos
      Abstract: Chronic kidney disease (CKD) is a worldwide public health problem that affects millions of men and women of all ages and racial groups. Loss of mesangial cells (MC) represents an early common feature in the pathogenesis of CKD. Transforming growth factor-β1 (TGF-β1) is a key inducer of kidney damage and triggers several pathological changes in renal cells, notably MC apoptosis. However, the mechanism of MC apoptosis induced by TGF-β1 remains elusive. Here, we demonstrate for the first time a novel regulatory pathway in which the disheveled-binding antagonist of β-catenin 1 (Dact1) gene is upregulated by TGF-β1, inducing MC apoptosis. We also show that the inhibitory effect of Dact1 and TGF-β1 on the transcriptional activation of the pro-survival Wnt pathway is the mechanism of death induction. In addition, Dact1 mRNA/protein levels are increased in kidney remnants from 5/6 nephrectomized rats and strongly correlate with TGF-β1 expression. Together, our results point to Dact1 as a novel element controlling MC survival that is causally related to CKD progression. J. Cell. Physiol. 9999: 1–8, 2016. © 2016 Wiley Periodicals, Inc.Here, we demonstrate for the first time a novel regulatory pathway in which the disheveled-binding antagonist of β-catenin 1 (Dact1) gene is upregulated by TGF-β1, inducing MC apoptosis.
      PubDate: 2017-02-16T09:46:07.825522-05:
      DOI: 10.1002/jcp.25636
       
  • Proteomics Applications in Dental Derived Stem Cells
    • Authors: Jie Li; Weidong Tian, Jinlin Song
      Abstract: At present, the existence of a variety of dental derived stem cells has been documented. These cells displayed promising clinical application potential not only for teeth and its surrounding tissue regeneration, but also for other tissues, such as nerve and bone regeneration. Proteomics is an unbiased, global informatics tool that provides information on all protein expression levels as well as post-translational modification in cells or tissues and is applicable to dental derived stem cells research. Over the last decade, considerable progress has been made to study the global proteome, secrotome, and membrane proteome of dental derived stem cells. Here, we present an overview of the proteomics studies in the context of stem cell research. Particular attention is given to dental derived stem cell types as well as current challenges and opportunities. J. Cell. Physiol. 9999: 1–9, 2016. © 2016 Wiley Periodicals, Inc.(1) Dental derived stem cells displayed promising clinical application potential not only for teeth and its surrounding tissue regeneration, but also for other tissues regeneration. (2) Here, we present an overview of the proteomics studies in the context of dental derived stem cell research. (3) Particular attention is given to dental derived stem cell types as well as current challenges and opportunities.
      PubDate: 2017-02-16T09:40:25.198627-05:
      DOI: 10.1002/jcp.25667
       
  • Tight Junction Protein Abundance and Apoptosis During Involution of Rat
           Mammary Glands
    • Authors: Claire V. C. Phyn; Kerst Stelwagen, Stephen R. Davis, Christopher D. McMahon, Joanne M. Dobson, Kuljeet Singh
      Abstract: To examine tight junction protein abundance and apoptosis of epithelial cells at the onset of involution in rodent mammary glands, milk accumulation and mammary engorgement were induced by teat-sealing with an adhesive for 0, 6, 12, 18, 24, and 36 h (n = 6 per group) at peak lactation. In non-sealed control glands, histological analysis confirmed a lactating phenotype, indicating suckling by pups throughout the experiment. In contrast, alveoli of teat-sealed glands were distended within 6 h, with maximal luminal size observed by 12 h of non-suckling. By 18 h following teat-sealing, an involuting phenotype was observed, indicated by alveolar lumina engorged with milk vesicles and increased leukocytes. Relative to non-sealed glands, mammary apoptosis was increased in engorged glands 18 h following teat-sealing. The abundance of ZO-1 and occludin proteins was decreased in engorged glands by 12 and 18 h, respectively, following teat-sealing. In contrast, the claudin-1 22 kDa band was increased by 6 h and peaked at 12–18 h, whereas the 28 kDa band declined by 36 h, relative to controls. There were no temporal changes in ZO-1, occludin, and claudin-1 22 kDa proteins within control glands, although there were minor differences in claudin-1 28 kDa. These data indicate that intramammary milk accumulation due to cessation of milk removal is associated with mammary apoptosis. The apoptotic event is preceded by a rapid loss of abundance of ZO-1, occludin and an initial increase in claudin-1. The loss of cell–cell communication may initiate involution and apoptosis of mammary epithelial cells and is a localized intramammary event, occurring only in non-suckled glands. J. Cell. Physiol. 9999: 1–8, 2016. © 2016 Wiley Periodicals, Inc.To examine tight junction protein abundance and apoptosis of epithelial cells at the onset of involution in rodent mammary glands, milk accumulation and mammary engorgement was induced by teat-sealing for 0, 6, 12, 18, 24, and 36 h (n = 6 per group) at peak lactation. The loss of cell–cell communication may initiate involution and apoptosis of mammary epithelial cells and is a localized intramammary event, occurring only in non-suckled glands.
      PubDate: 2017-02-16T09:30:26.601031-05:
      DOI: 10.1002/jcp.25591
       
  • Protective Role of GPER Agonist G-1 on Cardiotoxicity Induced by
           Doxorubicin
    • Authors: Ernestina M. De Francesco; Carmine Rocca, Francesco Scavello, Daniela Amelio, Teresa Pasqua, Damiano C. Rigiracciolo, Andrea Scarpelli, Silvia Avino, Francesca Cirillo, Nicola Amodio, Maria C. Cerra, Marcello Maggiolini, Tommaso Angelone
      Abstract: The use of Doxorubicin (Dox), a frontline drug for many cancers, is often complicated by dose-limiting cardiotoxicity in approximately 20% of patients. The G-protein estrogen receptor GPER/GPR30 mediates estrogen action as the cardioprotection under certain stressful conditions. For instance, GPER activation by the selective agonist G-1 reduced myocardial inflammation, improved immunosuppression, triggered pro-survival signaling cascades, improved myocardial mechanical performance, and reduced infarct size after ischemia/reperfusion (I/R) injury. Hence, we evaluated whether ligand-activated GPER may exert cardioprotection in male rats chronically treated with Dox. 1 week of G-1 (50 μg/kg/day) intraperitoneal administration mitigated Dox (3 mg/kg/day) adverse effects, as revealed by reduced TNF-α, IL-1β, LDH, and ROS levels. Western blotting analysis of cardiac homogenates indicated that G-1 prevents the increase in p-c-jun, BAX, CTGF, iNOS, and COX2 expression induced by Dox. Moreover, the activation of GPER rescued the inhibitory action elicited by Dox on the expression of BCL2, pERK, and pAKT. TUNEL assay indicated that GPER activation may also attenuate the cardiomyocyte apoptosis upon Dox exposure. Using ex vivo Langendorff perfused heart technique, we also found an increased systolic recovery and a reduction of both infarct size and LDH levels in rats treated with G-1 in combination with Dox respect to animals treated with Dox alone. Accordingly, the beneficial effects induced by G-1 were abrogated in the presence of the GPER selective antagonist G15. These data suggest that GPER activation mitigates Dox-induced cardiotoxicity, thus proposing GPER as a novel pharmacological target to limit the detrimental cardiac effects of Dox treatment. J. Cell. Physiol. 9999: 1–10 , 2016. © 2016 Wiley Periodicals, Inc.1. The potential beneficial effects of GPER activation in Doxorubicin (Dox)-induced cardiotoxicity was evaluated. 2. In male rat heart, the treatment with the selective GPER ligand G-1 prevented the increase in TNF-α, IL-1β, and LDH plasma levels induced by Dox. In Langendorff perfused rat heart, G-1 also increased the systolic recovery at the end of ischemia and reduced infarct size and LDH levels triggered by Dox. Moreover, G-1 attenuated the Dox-induced myocyte apoptosis, as well as the inhibition of pro-survival signaling cascades like ERK1/2 and AKT. Next, G-1 counteracted the down-regulation of BCL2 as well as the up-regulation of apoptotic, inflammatory and fibrotic targets, such as BAX, iNOS, COX2, and CTGF induced by Dox. 3. Our study paves the way for further analyzing the role of GPER as pharmacological target in novel therapeutic strategies aimed to prevent the cardiotoxicity exerted by Dox.
      PubDate: 2017-02-16T09:20:30.356957-05:
      DOI: 10.1002/jcp.25585
       
  • Postnatal Hyperplasic Effects of ActRIIB Blockade in a Severely Dystrophic
           Muscle
    • Authors: Cory Nielsen; Ross M. Potter, Christopher Borowy, Kimberly Jacinto, Ravi Kumar, C. George Carlson
      Abstract: The efficacy of two ActRIIB ligand-trapping agents (RAP-031 and RAP-435) in treating muscular dystrophy was examined by determining their morphological effects on the severely dystrophic triangularis sterni (TS) muscle of the mdx mouse, a model for Duchenne muscular dystrophy. These agents trap all endogenous ligands to the ActRIIB receptor and thereby block myostatin signaling in a highly selective manner. Short-term (1 month) and long-term (3 months) in vivo treatment of 1-month-old mdx mice increased myonuclei and fiber cross section (FCS) density but did not alter individual fiber size. Vehicle-treated mdx mice exhibited age-dependent increases in myonuclei and FCS density, and age-dependent reductions in centronucleation that were each enhanced by treatment with RAP-435. Distributions of FCS area (FCSA) in the mdx TS were 90% identical to those from untreated age-matched nondystrophic mice and were unaltered by the substantial fiber hyperplasia observed with age and RAP-435 treatment. These results were inconsistent with injury-induced fiber regeneration which produces altered FCSA distributions characterized by a distinct class of smaller regenerated fibers. Nondystrophic mice exhibited a constant postnatal density of fiber cross sections and myonuclei, and RAP-435 treatment of nondystrophic mice increased TS mean FCSA but had no effects on myonuclei or FCS density. These results demonstrating a continual postnatal proliferation and fusion of satellite cells and a response to myostatin blockade characteristic of developing prenatal muscle suggest that the lack of dystrophin directly results in unrestrained postnatal satellite cell activation that is not necessarily dependent upon prior fiber degeneration. J. Cell. Physiol. 9999: 1–20, 2016. © 2016 Wiley Periodicals, Inc.We examined the efficacy of two ActRIIB ligand-trapping agents in altering the morphology of a severely dystrophic muscle in the mdx mouse. The results indicate that severely dystrophic muscle exhibits monotonic postnatal increases in myonuclei and fiber density that are not associated with alterations in the distribution of fiber cross-sectional areas. Long-term treatment with an ActRIIB ligand-trapping agent substantially enhanced this postnatal hyperplasia and reduced centronucleation. These results demonstrating a continual postnatal proliferation and fusion of satellite cells and a response to myostatin blockade characteristic of developing prenatal muscle suggest that the lack of dystrophin directly results in unrestrained postnatal satellite cell activation that is not necessarily dependent upon prior fiber degeneration.
      PubDate: 2017-02-16T09:15:56.755686-05:
      DOI: 10.1002/jcp.25694
       
  • Role of Wnt5a in the Pathogenesis of Inflammatory Diseases
    • Authors: Mehran Pashirzad; Mojtaba Shafiee, Farzad Rahmani, Reihane Behnam-Rassouli, Fatemeh Hoseinkhani, Mikhail Ryzhikov, Maryam Moradi Binabaj, Mohammad Reza Parizadeh, Amir Avan, Seyed Mahdi Hassanian
      Abstract: Wnt5a initiates pro-inflammatory responses through activation of non-canonical Wnt signaling pathway. Pro-inflammatory functions of Wnt5a trigger pro-inflammatory signaling cascades and increase secretion of pro-inflammatory cytokines and chemokines. Wnt5a as a potent signaling molecule is strongly implicated in a number of diseases including cancer, diabetes, metabolic disorders, and of special interest in this review, inflammatory diseases. This review summarizes the role of Wnt5a in the pathogenesis of inflammatory diseases including atherosclerosis, rheumatoid arthritis, psoriasis vulgaris and sepsis, promoting greater understanding, and clinical management of these diseases. J. Cell. Physiol. 9999: 1–6, 2017. © 2016 Wiley Periodicals, Inc.Wnt5a as a potent signaling molecule is strongly implicated in a number of diseases including cancer, diabetes, metabolic disorders, and of special interest in this review, inflammatory diseases. This review summarizes the role of Wnt5a in the pathogenesis of inflammatory diseases including atherosclerosis, rheumatoid arthritis, psoriasis vulgaris and sepsis, promoting greater understanding and clinical management of these diseases.
      PubDate: 2017-02-16T09:10:27.206473-05:
      DOI: 10.1002/jcp.25687
       
  • M2-like macrophages induce colon cancer cell invasion via matrix
           metalloproteinases
    • Authors: Katyayni Vinnakota; Yuan Zhang, Benson Chellakkan Selvanesan, Geriolda Topi, Tavga Salim, Janna Sand-Dejmek, Gunilla Jönsson, Anita Sjölander
      Abstract: The inflammatory milieu plays an important role in colon cancer development and progression. Previously, we have shown that tumor-associated macrophages (TAMs), an important component of the tumor microenvironment, are enriched in tumors compared with normal tissue and confer a poorer prognosis. In the present study, we found that matrix metallopeptidase-9 (MMP-9), which degrades extracellular matrix proteins, was increased in biopsies from colon cancer patients and in mouse xenografts with SW480 cell-derived tumors. SW480 colon cancer cells exposed to M2-like macrophage-conditioned medium (M2-medium) exhibited increased MMP-9 mRNA, protein expression and gelatinase activity. A similar effect was obtained by the addition of tumor necrosis factor-α (TNFα) and leukotriene D4 (LTD4). MMP-9 expression and activity were reduced by a TNFα blocking antibody adalimumab and a cysteinyl leukotriene receptor 1 (CysLTR1, the receptor for LTD4) antagonist montelukast. M2-medium also induced changes in the epithelial–mesenchymal transition (EMT) markers E-cadherin, β-catenin, vimentin, and snail in SW480 cells. We also found that both M2-medium and TNFα and LTD4 induced stabilization/nuclear translocation of β-catenin. Furthermore, we also observed an elongated phenotype that may indicate increased invasiveness, as confirmed in a collagen I invasion assay. M2-medium increased the invasive ability, and a similar effect was also obtained by the addition of TNFα and LTD4. The specific MMP inhibitor I or adalimumab and montelukast reduced the number of invasive cells. In conclusion, our findings show that M2-medium enriched in TNFα and LTD4 promote colon cancer cell invasion via MMP-9 expression and activation and the induction of EMT.Our findings show that M2-medium enriched in TNFalfa and LTD4 promote colon cancer cell invasion via MMP-9 expression and activation and the induction of EMT in SW480 colon cancer cells.
      PubDate: 2017-02-13T12:00:33.599029-05:
      DOI: 10.1002/jcp.25808
       
  • Knockdown of REV3 synergizes with ATR inhibition to promote apoptosis
           induced by cisplatin in lung cancer cells
    • Authors: He-Guo Jiang; Ping Chen, Jin-Yu Su, Ming Wu, Hai Qian, Yi Wang, Jian Li
      Abstract: It has been demonstrated that REV3, the catalytic subunit of the translesion synthesis (TLS) polymerase ζ, play an important role in DNA damage response (DDR) induced by cisplatin, and Ataxia-telangietasia mutated and Rad-3-related (ATR) knase is a central player in activating cell cycle checkpoint, stabilizing replication forks, regulating DDR, and promoting repair of DNA damage caused by cisplatin. Cancer cells deficient in either one of REV3 and ATR are more sensitive to cisplatin. However, whether co-inhibition of REV3 and ATR can further increase sensitivity of non-small cell lung cancer (NSCLC) cells to cisplatin is not clear. In this study, we show that REV3 knockdown combined with ATR inhibition further enhance cytotoxicity of cisplatin in NSCLC cells, including cisplatin-sensitive and -resistant cell lines, compared to individual knockdown of REV3 or ATR, which are accompanied by markedly caspase-dependent apoptosis response, pronounced DNA damage accumulation and severe impediment of interstrand crosslink (ICL), and double strand break (DSB) repair. Our results suggest that REV3 knockdown synergize strongly with ATR inhibition to significantly increase sensitivity of cisplatin in NSCLC cells by inhibiting ICL and DSB repair. Thus simultaneously targeting REV3 and ATR may represent one approach to overcome cisplatin resistance and improve chemotherapeutic efficacy in NSCLC treatment.Herein, we showed that depletion of REV3 combined with ATR inhibition significantly potentiates the cytotoxicity of cisplatin to non-small cell lung cancer (NSCLC) cells, which is concomitant with pronounced DNA damage accumulation and severe impediment of interstrand crosslink (ICL) and double strand break (DSB) repair, compared to individual depletion of REV3 or ATR. Our data demonstrate that REV3 depletion can synergize with ATR inhibition to increase suppression effect of cisplatin on NSCLC cells through inhibition of ICL and DSB repair.
      PubDate: 2017-02-13T11:35:41.724322-05:
      DOI: 10.1002/jcp.25792
       
  • Lipid profiling of parkin-mutant human skin fibroblasts
    • Authors: Simona Lobasso; Paola Tanzarella, Daniele Vergara, Michele Maffia, Tiziana Cocco, Angela Corcelli
      Abstract: Parkin mutations are a major cause of early-onset Parkinson's disease (PD). The impairment of protein quality control system together with defects in mitochondria and autophagy process are consequences of the lack of parkin, which leads to neurodegeneration. Little is known about the role of lipids in these alterations of cell functions. In the present study, parkin-mutant human skin primary fibroblasts have been considered as cellular model of PD to investigate on possible lipid alterations associated with the lack of parkin protein. Dermal fibroblasts were obtained from two unrelated PD patients with different parkin mutations and their lipid compositions were compared with that of two control fibroblasts. The lipid extracts of fibroblasts have been analyzed by combined matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry (MALDI-TOF/MS) and thin-layer chromatography (TLC). In parallel, we have performed direct MALDI-TOF/MS lipid analyses of intact fibroblasts by skipping lipid extraction steps. Results show that the proportions of some phospholipids and glycosphingolipids were altered in the lipid profiles of parkin-mutant fibroblasts. The detected higher level of gangliosides, phosphatidylinositol, and phosphatidylserine could be linked to dysfunction of autophagy and mitochondrial turnover; in addition, the lysophosphatidylcholine increase could represent the marker of neuroinflammatory state, a well-known component of PD.In the present study, parkin-mutant human skin primary fibroblasts have been considered as cellular model of Parkinson's disease (PD) in order to investigate on possible lipid alterations associated with the lack of parkin protein. Dermal fibroblasts were obtained from two unrelated PD patients with different parkin mutations and their lipid compositions were compared with that of two control fibroblasts. The lipid extracts of fibroblasts have been analyzed by combined MALDI-TOF/MS and TLC. In parallel, we have performed direct MALDI-TOF/MS lipid analyses of intact fibroblasts. Results show that the proportions of some phospholipids and glycosphingolipids were altered in the lipid profiles of parkin-mutant fibroblasts.
      PubDate: 2017-02-10T16:00:26.702925-05:
      DOI: 10.1002/jcp.25815
       
  • Mesenchymal stem cells improves survival in LPS-induced acute lung injury
           acting through inhibition of NETs formation
    • Authors: Leonardo Pedrazza; Aline Andrea Cunha, Carolina Luft, Nailê Karine Nunes, Felipe Schimitz, Rodrigo Benedetti Gassen, Ricardo Vaz Breda, Marcio Vinícius Fagundes Donadio, Angela Terezinha de Souza Wyse, Paulo Marcio Condessa Pitrez, Jose Luis Rosa, Jarbas Rodrigues de Oliveira
      Abstract: Acute lung injury (ALI) and acute respiratory distress syndrome (ARDS) are syndromes of acute hypoxemic respiratory failure resulting from a variety of direct and indirect injuries to the gas exchange parenchyma of the lungs. During the ALI, we have an increase release of proinflammatory cytokines and high reactive oxygen species (ROS) formation. These factors are responsible for the release and activation of neutrophil-derived proteases and the formation of neutrophil extracellular traps (NETs). The excessive increase in the release of NETs cause damage to lung tissue. Recent studies have studies involving the administration of mesenchymal stem cells (MSCs) for the treatment of experimental ALI has shown promising results. In this way, the objective of our study is to evaluate the ability of MSCs, in a lipopolysaccharide (LPS)-induced ALI model, to reduce inflammation, oxidative damage, and consequently decrease the release of NETs. Mice were submitted lung injury induced by intratracheal instillation of LPS and subsequently treated or not with MSCs. Treatment with MSCs was able to modulate pulmonary inflammation, decrease oxidative damage, and reduce the release of NETs. These benefits from treatment are evident when we observe a significant increase in the survival curve in the treated animals. Our results demonstrate that MSCs treatment is effective for the treatment of ALI. For the first time, it is described that MSCs can reduce the formation of NETs and an experimental model of ALI. This finding is directly related to these cells modulate the inflammatory response and oxidative damage in the course of the pathology.Treatment with MSCs was able to modulate pulmonary inflammation, decrease oxidative damage, and reduce the release of NETs. For the first time, it is described that MSCs can reduce the formation of NETs and an experimental model of ALI.
      PubDate: 2017-02-09T10:56:10.426812-05:
      DOI: 10.1002/jcp.25816
       
  • Modulation of long-term endothelial-barrier integrity is conditional to
           the cross-talk between Akt and Src signaling
    • Authors: Fei Gao; Harika Sabbineni, Sandeep Artham, Payaningal R. Somanath
      Abstract: Although numerous studies have implicated Akt and Src kinases in vascular endothelial growth factor (VEGF) and Angiopoietin-1 (Ang-1)-induced endothelial-barrier regulation, a link between these two pathways has never been demonstrated. We determined the long-term effects of Akt inhibition on Src activity and vice versa, and in turn, on the human microvascular endothelial cell (HMEC) barrier integrity at the basal level, and in response to growth factors. Our data showed that Akt1 gene knockdown increases gap formation in HMEC monolayer at the basal level. Pharmacological inhibition of Akt, but not Src resulted in exacerbated VEGF-induced vascular leakage and impaired Ang-1-induced HMEC-barrier protection in vitro at 24 hr. Whereas inhibition of Akt had no effect on VEGF-induced HMEC gap formation in the short term, inhibition of Src blunted this process. In contrast, inhibition of Akt disrupted the VEGF and Ang-1 stabilized barrier integrity in the long-term while inhibition of Src did not. Interestingly, both long-term Akt inhibition and Akt1 gene knockdown in HMECs resulted in increased Tyr416 phosphorylation of Src. Treatment of HMECs with transforming growth factor-β1 (TGFβ1) that inhibited Akt Ser473 phosphorylation in the long-term, activated Src through increased Tyr416 phosphorylation and decreased HMEC-barrier resistance. The effect of TGFβ1 on endothelial-barrier breakdown was blunted in Akt1 deficient HMEC monolayers, where endothelial-barrier resistance was already impaired compared to the control. To our knowledge, this is the first report demonstrating a direct cross-talk between Akt and Src in endothelial-barrier regulation.The current study identifies the long-term effects of VEGF and Ang-1 on HMEC-barrier protection via increased Akt and normalized Src activities, and that sustained treatment with TGFβ1 promotes endothelial-barrier breakdown associated with inhibition of Akt and activation of Src. Direct inhibition of Akt activity using triciribine or gene silencing lead to Src activation in the long-term indicating that Akt and Src are reciprocally regulated in the long-term endothelial-barrier function.
      PubDate: 2017-02-09T10:51:15.187846-05:
      DOI: 10.1002/jcp.25791
       
  • Impact of lysosomal storage disorders on biology of mesenchymal stem
           cells: Evidences from in vitro silencing of glucocerebrosidase (GBA) and
           alpha-galactosidase A (GLA) enzymes
    • Authors: Tiziana Squillaro; Ivana Antonucci, Nicola Alessio, Anna Esposito, Marilena Cipollaro, Mariarosa Anna Beatrice Melone, Gianfranco Peluso, Liborio Stuppia, Umberto Galderisi
      Abstract: Lysosomal storage disorders (LDS) comprise a group of rare multisystemic diseases resulting from inherited gene mutations that impair lysosomal homeostasis. The most common LSDs, Gaucher disease (GD), and Fabry disease (FD) are caused by deficiencies in the lysosomal glucocerebrosidase (GBA) and alpha-galactosidase A (GLA) enzymes, respectively. Given the systemic nature of enzyme deficiency, we hypothesized that the stem cell compartment of GD and FD patients might be also affected. Among stem cells, mesenchymal stem cells (MSCs) are a commonly investigated population given their role in hematopoiesis and the homeostatic maintenance of many organs and tissues. Since the impairment of MSC functions could pose profound consequences on body physiology, we evaluated whether GBA and GLA silencing could affect the biology of MSCs isolated from bone marrow and amniotic fluid. Those cell populations were chosen given the former's key role in organ physiology and the latter's intriguing potential as an alternative stem cell model for human genetic disease. Our results revealed that GBA and GLA deficiencies prompted cell cycle arrest along with the impairment of autophagic flux and an increase of apoptotic and senescent cell percentages. Moreover, an increase in ataxia–telangiectasia-mutated staining 1 hr after oxidative stress induction and a return to basal level at 48 hr, along with persistent gamma-H2AX staining, indicated that MSCs properly activated DNA repair signaling, though some damages remained unrepaired. Our data therefore suggest that MSCs with reduced GBA or GLA activity are prone to apoptosis and senescence due to impaired autophagy and DNA repair capacity.Gaucher disease (GD) and Fabry disease (FD) are caused by deficiencies in the lysosomal glucocerebrosidase (GBA) and alpha-galactosidase A (GLA) enzymes, respectively. Given the systemic nature of enzyme deficiency, we hypothesized that the stem cell compartment of GD and FD patients might be also affected; thus, we evaluated whether GBA and GLA silencing could affect the biology of mesenchymal stem cells (MSCs) isolated from bone marrow and amniotic fluid. Our data suggest that MSCs with reduced GBA or GLA activity are prone to apoptosis and senescence due to impaired autophagy and DNA repair capacity.
      PubDate: 2017-02-07T10:55:32.192299-05:
      DOI: 10.1002/jcp.25807
       
  • LSP1, a responsive protein from Meyerozyma guilliermondii, elicits defence
           response and improves glycyrrhizic acid biosynthesis in Glycyrrhiza
           uralensis Fisch adventitious roots
    • Authors: Juan Wang; Jianli Li, Jing Li, Jinxin Li, Shujie Liu, Wenyuan Gao
      Abstract: This research explored the effects of protein and polysaccharide in Meyerozyma guilliermondii on active compounds in Glycyrrhiza uralensis Fisch adventitious roots. In this study, a responsive protein LSP1 was purified from the Meyerozyma guilliermondii since the excellent induction. The contents of total flavonoids (3.46 mg · g−1), glycyrrhizic acid (0.41 mg · g−1), glycyrrhetinic acid (0.41 mg · g−1), and polysaccharide (94.49 mg · g−1) in adventitious root peaked at LSP1 group, which were 1.6, 3.4, 2.4, 2.0-fold that of control, respectively. Besides, the responsive protein LSP1 significantly activated the defense signaling, mitogen-activated protein kinases and extremely up-regulated the expression of defense-related genes and functional genes involved in glycyrrhizic acid biosynthesis.This research explored the effects of protein and polysaccharide in Meyerozyma guilliermondii on active compounds in Glycyrrhiza uralensis Fisch adventitious roots. In this study, a responsive protein LSP1 was purified from the M. guilliermondii since the excellent induction. Besides, the responsive protein LSP1 significantly activated the defense signaling, mitogen-activated protein kinases and extremely up-regulated the expression of defense-related genes and functional genes involved in glycyrrhizic acid biosynthesis.
      PubDate: 2017-02-07T10:45:28.98882-05:0
      DOI: 10.1002/jcp.25811
       
  • Involvement of CX3CL1 in the Migration of Osteoclast Precursors Across
           Osteoblast Layer Stimulated by Interleukin-1ß
    • Authors: Tsuyoshi Matsuura; Shizuko Ichinose, Masako Akiyama, Yuki Kasahara, Noriko Tachikawa, Ken-ichi Nakahama
      Abstract: The trigger for bone remodeling is bone resorption by osteoclasts. Osteoclast differentiation only occurs on the old bone, which needs to be repaired under physiological conditions. However, uncontrolled bone resorption is often observed in pro-inflammatory bone diseases, such as rheumatoid arthritis. Mature osteoclasts are multinuclear cells that differentiate from monocyte/macrophage lineage cells by cell fusion. Although Osteoclast precursors should migrate across osteoblast layer to reach bone matrix before maturation, the underlying mechanisms have not yet been elucidated in detail. We herein found that osteoclast precursors utilize two routes to migrate across osteoblast layer by confocal- and electro-microscopic observations. The osteoclast supporting activity of osteoblasts inversely correlated with osteoblast density and was positively related to the number of osteoclast precursors under the osteoblast layer. Osteoclast differentiation was induced by IL-1ß, but not by PGE2 in high-density osteoblasts. Osteoblasts and osteoclast precursors expressed CX3CL1 and CX3CR1, respectively, and the expression of CX3CL1 increased in response to interleukin-1ß. An anti-CX3CL1-neutralizing antibody inhibited the migration of osteoclast precursors and osteoclast differentiation. These results strongly suggest the involvement of CX3CL1 in the migration of osteoclast precursors and osteoclastogenesis, and will contribute to the development of new therapies for bone diseases. J. Cell. Physiol. 9999: 1–7, 2016. © 2016 Wiley Periodicals, Inc.Transcellular and paracellular migration of osteoclast precursors across osteoblast layer were observed. CX3CL1 was involved in the migration of osteoclast precursors and osteoclastogenesis.
      PubDate: 2017-02-03T10:00:30.140846-05:
      DOI: 10.1002/jcp.25577
       
  • Membrane ion Channels and Receptors in Animal lifespan Modulation
    • Authors: Yi Sheng; Lanlan Tang, Lijun Kang, Rui Xiao
      Abstract: Acting in the interfaces between environment and membrane compartments, membrane ion channels, and receptors transduce various physical and chemical cues into downstream signaling events. Not surprisingly, these membrane proteins play essential roles in a wide range of cellular processes such as sensory perception, synaptic transmission, cellular growth and development, fate determination, and apoptosis. However, except insulin and insulin-like growth factor receptors, the functions of membrane receptors in animal lifespan modulation have not been well appreciated. On the other hand, although ion channels are popular therapeutic targets for many age-related diseases, their potential roles in aging itself are largely neglected. In this review, we will discuss our current understanding of the conserved functions and mechanisms of membrane ion channels and receptors in the modulation of lifespan across multiple species including Caenorhabditis elegans, Drosophila, mouse, and human.Acting in the interfaces between environment and membrane compartments, membrane ion channels and receptors transduce various physical and chemical cues into downstream signaling events. In this review, we will discuss our current understanding of the conserved functions and mechanisms of membrane ion channels, and receptors in the modulation of lifespan across multiple species including Caenorhabditis elegans, Drosophila, mouse, and human.
      PubDate: 2017-02-03T09:55:26.623235-05:
      DOI: 10.1002/jcp.25824
       
  • DNA Methylation Profiling in Chondrocyte Dedifferentiation In Vitro
    • Authors: Li Duan; Yujie Liang, Bin Ma, Daming Wang, Wei Liu, Jianghong Huang, Jianyi Xiong, Liangquan Peng, Jielin Chen, Weimin Zhu, Daping Wang
      Abstract: DNA methylation has emerged as a crucial regulator of chondrocyte dedifferentiation, which severely compromises the outcome of autologous chondrocyte implantation (ACI) treatment for cartilage defects. However, the full-scale DNA methylation profiling in chondrocyte dedifferentiation remains to be determined. Here, we performed a genome-wide DNA methylation profiling of dedifferentiated chondrocytes in monolayer culture and chondrocytes treated with DNA methylation inhibitor 5-azacytidine (5-AzaC). This research revealed that the general methylation level of CpG was increased while the COL-1A1 promoter methylation level was decreased during the chondrocyte dedifferentiation. 5-AzaC could reduce general methylation levels and reverse the chondrocyte dedifferentiation. Surprisingly, the DNA methylation level of COL-1A1 promoter was increased after 5-AzaC treatment. The COL-1A1 expression level was increased while that of SOX-9 was decreased during the chondrocyte dedifferentiation. 5-AzaC treatment up-regulated the SOX-9 expression while down-regulated the COL-1A1 promoter activity and gene expression. Taken together, these results suggested that differential regulation of the DNA methylation level of cartilage-specific genes might contribute to the chondrocyte dedifferentiation. Thus, the epigenetic manipulation of these genes could be a potential strategy to counteract the chondrocyte dedifferentiation accompanying in vitro propagation. J. Cell. Physiol. 9999: 1–9, 2016. © 2016 Wiley Periodicals, Inc.Differential regulation of the DNA methylation level of cartilage-specific genes might contribute the chondrocyte dedifferentiation. Thus, the epigenetic manipulation of these genes could be a potential strategy to counteract the chondrocyte dedifferentiation accompanying in vitro propagation.
      PubDate: 2017-01-31T07:47:23.34891-05:0
      DOI: 10.1002/jcp.25486
       
  • Glioblastoma Multiforme and Adult Neurogenesis in the
           Ventricular-Subventricular Zone: A Review
    • Authors: Claudia Capdevila; Lucía Rodríguez Vázquez, Joaquín Martí
      Abstract: Brain cancers account for
      PubDate: 2017-01-31T07:47:08.85698-05:0
      DOI: 10.1002/jcp.25502
       
  • Cytoskeleton Aberrations in Alkaptonuric Chondrocytes
    • Authors: Michela Geminiani; Silvia Gambassi, Lia Millucci, Pietro Lupetti, Giulia Collodel, Lucia Mazzi, Bruno Frediani, Daniela Braconi, Barbara Marzocchi, Marcella Laschi, Giulia Bernardini, Annalisa Santucci
      Abstract: Alkaptonuria (AKU) is an ultra-rare autosomal genetic disorder caused by a defect in the activity of the enzyme homogentisate 1,2-dioxygenase (HGD) that leads to the accumulation of homogentisic acid (HGA) and its oxidized product, benzoquinone acetic acid (BQA), in the connective tissues causing a pigmentation called “ochronosis.” The consequent progressive formation of ochronotic aggregates generate a severe condition of oxidative stress and inflammation in all the affected areas. Experimental evidences have also proved the presence of serum amyloid A (SAA) in several AKU tissues and it allowed classifying AKU as a secondary amyloidosis. Although AKU is a multisystemic disease, the most affected system is the osteoarticular one and articular cartilage is the most damaged tissue. In this work, we have analyzed for the first time the cytoskeleton of AKU chondrocytes by means of immunofluorescence staining. We have shown the presence of SAA within AKU chondrocytes and finally we have demonstrated the co-localization of SAA with three cytoskeletal proteins: actin, vimentin, and β-tubulin. Furthermore, in order to observe the ultrastructural features of AKU chondrocytes we have performed TEM analysis, focusing on the Golgi apparatus structure and, to demonstrate that pigmented areas in AKU cartilage are correspondent to areas of oxidation, 4-HNE presence has been evaluated by means of immunofluorescence. J. Cell. Physiol. 9999: 1–11, 2016. © 2016 Wiley Periodicals, Inc.The cytoskeleton of alkaptonuric chondrocytes is severely alterated. Serum amyloid A is demonstrated to be present within alkaptonuric chondrocytes and to co-localize with the three main cytoskeletal proteins: actin, vimentin, and β-tubulin. These findings lead to hypothesize a role of SAA in cytoskeleton alterations of alkaptonuric chondrocytes.
      PubDate: 2017-01-31T07:29:41.564847-05:
      DOI: 10.1002/jcp.25500
       
  • Table of Contents, Editor's Choice, Highlights
    • Pages: 1217 - 1224
      PubDate: 2017-02-24T12:13:54.811026-05:
      DOI: 10.1002/jcp.25833
       
  • A Novel Skin Splint for Accurately Mapping Dermal Remodeling and
           Epithelialization During Wound Healing
    • Authors: Shiro Jimi; Francesco De Francesco, Giuseppe A. Ferraro, Michele Riccio, Shuuji Hara
      Pages: 1225 - 1232
      Abstract: The mouse excisional dorsal full-thickness wound model with a silicon splint fixed on the skin has been widely used to mimic human wound healing. However, the method cannot accurately quantify dermal remodeling, since the initial point of epithelialization on the wound surface is unclear. To overcome this limitation, we have developed a novel mouse excisional wound model to assess the degree of epithelial extension and regeneration, using a plastic ring-shaped splint fixed beneath the surrounding epidermal tissue. At the end of the experiment, tissue samples were fixed in formalin, the splint was excised, and paraffin sections were prepared. Splint holes, corresponding to the prior location of the splint, were evident on the tissue cross-sections, and the epidermis above the holes was considered the initial excision site. The epidermal contraction and epithelial regeneration, as independent essential tissue alterations in wound healing, could be distinguishable and quantified. Compared with previous splint models, this method provides an accurate evaluation of epidermal processes in wound healing, and can be a platform to assess the effects of various wound healing factors. J. Cell. Physiol. 232: 1225–1232, 2017. © 2016 Wiley Periodicals, Inc.The new method established in this manuscript makes possible the accurate assessment and measurement of epidermal changes—dermal remodeling and regeneration—in the wound healing process in mice.
      PubDate: 2017-01-06T10:15:32.764507-05:
      DOI: 10.1002/jcp.25595
       
  • The Emerging Role of TRAF7 in Tumor Development
    • Authors: Tiziana Zotti; Ivan Scudiero, Pasquale Vito, Romania Stilo
      Pages: 1233 - 1238
      Abstract: The seven members of the tumor necrosis factor receptor (TNF-R)-associated factor (TRAF) family of intracellular proteins were originally discovered and characterized as signaling adaptor molecules coupled to the cytoplasmic regions of receptors of the TNF-R superfamily. Functionally, TRAFs act both as a scaffold and/or enzymatic proteins to regulate activation of mitogen-activated protein kinases (MAPKs) and transcription factors of nuclear factor-κB family (NF-κB). Given the wide variety of stimuli intracellularly conveyed by TRAF proteins, they are physiologically involved in multiple biological processes, including embryonic development, tissue homeostasis, and regulation of innate and adaptive immune responses. In the last few years, it has become increasingly evident the involvement of TRAF7, the last member of the TRAF family to be discovered, in the genesis and progression of several human cancers, placing TRAF7 in the spotlight as a novel tumor suppressor protein. In this paper, we review and discuss the literature recently produced on this subject. J. Cell. Physiol. 232: 1233–1238, 2017. © 2016 The
      Authors . Journal of Cellular Physiology Published by Wiley Periodicals, Inc.TRAF proteins are signaling adaptor molecules that regulate activation of mitogen-activated protein kinases (MAPKs) and transcription factors of nuclear factor-κB family (NF-κB). In this paper, we review and discuss the literature recently produced pointing to an involvement of TRAF7 in the genesis and progression of several human cancers.
      PubDate: 2017-01-06T10:15:36.42382-05:0
      DOI: 10.1002/jcp.25676
       
  • Roles of Multifunctional COP9 Signalosome Complex in Cell Fate and
           Implications for Drug Discovery
    • Authors: Ping Li; Longxiang Xie, Yinzhong Gu, Jiang Li, Jianping Xie
      Pages: 1246 - 1253
      Abstract: The eight subunits containing COP9 signalosome (CSN) complex, is highly conserved among eukaryotes. CSN, identified as a negative regulator of photomorphogenesis, has also been demonstrated to be important in proteolysis, cellular signal transduction and cell cycle regulation in various eukaryotic organisms. This review mainly summarizes the roles of CSN in cell cycle regulation, signal transduction and apoptosis, and its potential as diagnostic biomarkers, drug targets for cancer and infectious diseases. J. Cell. Physiol. 232: 1246–1253, 2017. © 2016 Wiley Periodicals, Inc.This review mainly summarizes the roles of CSN in cell cycle regulation, signal transduction and apoptosis, and its potential as diagnostic biomarkers, drug targets for cancer and infectious diseases.
      PubDate: 2017-01-05T08:45:23.116809-05:
      DOI: 10.1002/jcp.25696
       
  • Unique Regulatory Mechanisms for the Human Embryonic Stem Cell Cycle
    • Authors: Jennifer J. VanOudenhove; Rodrigo A. Grandy, Prachi N. Ghule, Jane B. Lian, Janet L. Stein, Sayyed K. Zaidi, Gary S. Stein
      Pages: 1254 - 1257
      Abstract: The cell cycle in pluripotent human embryonic stem cells is governed by unique mechanisms that support unrestricted proliferation and competency for endodermal, mesodermal, and ectodermal differentiation. The abbreviated G1 period with retention of uncompromised fidelity for genetic and epigenetic mechanisms operative in control of proliferation support competency for expansion of the pluripotent cell population that is fundamental for initial stages of development. Regulatory events during the G1 period of the pluripotent cell cycle are decisive for the transition from pluripotency to lineage commitment. Recent findings indicate that a G2 cell cycle pause is present in both endodermal and mesodermal lineage cells, and is obligatory for differentiation to endoderm. J. Cell. Physiol. 232: 1254–1257, 2017. © 2016 Wiley Periodicals, Inc.The abbreviated G1 period with retention of uncompromised fidelity for genetic and epigenetic mechanisms operative in control of proliferation support competency for expansion of the pluripotent cell population that is fundamental for initial stages of development. Regulatory events during the G1 period of the pluripotent cell cycle are decisive for the transition from pluripotency to lineage commitment. Recent findings indicate that a G2 cell cycle pause is present in both endodermal and mesodermal lineage cells, and is obligatory for differentiation to endoderm.
      PubDate: 2017-01-31T07:36:36.861307-05:
      DOI: 10.1002/jcp.25567
       
  • Roles of Notch Signaling in Adipocyte Progenitor Cells and Mature
           Adipocytes
    • Authors: Tizhong Shan; Jiaqi Liu, Weiche Wu, Ziye Xu, Yizhen Wang
      Pages: 1258 - 1261
      Abstract: Adipose tissues, composed with mature adipocytes and preadipocytic stromal/stem cells, play crucial roles in whole body energy metabolism and regenerative medicine. Mature adipocytes are derived and differentiated from mesenchymal stem cells (MSCs) or preadipocytes. This differentiation process, also called adipogenesis, is regulated by several signaling pathways and transcription factors. Notch1 signaling is a highly conserved pathway that is indispensable for stem cell hemostasis and tissue development. In adipocyte progenitor cells, Notch1 signaling regulates the adipogenesis process including proliferation and differentiation of the adipocyte progenitor cells in vitro. Notably, the roles of Notch1 signaling in beige adipocytes formation, adipose development, and function, and the whole body energy metabolism have been recently reported. Here, we mainly review and discuss the roles of Notch1 signaling in adipogenesis in vitro as well as in beige adipocytes formation, adipocytes dedifferentiation, and function in vivo. J. Cell. Physiol. 232: 1258–1261, 2017. © 2016 Wiley Periodicals, Inc.Notch1 signaling not only regulates the adipogenesis process including proliferation and differentiation of the adipocyte progenitor cells in vitro, but also affects beige adipocytes formation, adipose development, and function, and the whole body energy metabolism in vivo.
      PubDate: 2017-01-05T08:50:23.366963-05:
      DOI: 10.1002/jcp.25697
       
  • Cyclic Tensile Strain Reduces TNF-α Induced Expression of MMP-13 by
           Condylar Temporomandibular Joint Cells
    • Authors: Hessam Tabeian; Astrid D. Bakker, Beatriz F. Betti, Frank Lobbezoo, Vincent Everts, Teun J. de Vries
      Pages: 1287 - 1294
      Abstract: To investigate whether the disproportionate degradation of mandibular condyle cartilage in arthritic juvenile temporomandibular joint (TMJ) is related to distinctive responses of TMJ-derived cells to tumor necrosis factor-α (TNF-α), and whether mechanical loading affects this response. The effect of TNF-α (0.1–10 ng/ml) was tested on juvenile porcine TMJ cells isolated from the condyle, fossa, and disc, grown in 3D agarose gels. Expression of anabolic and catabolic factors was quantified by RT-qPCR and/or immunohistochemistry. Condylar cells were stimulated for 12 h with TNF-α (10 ng/ml), followed by 8 h of 6% cyclic tensile strain, and gene expression of MMPs was quantified. TNF-α (10 ng/ml) reduced the expression of the matrix proteins collagen types I and II after 6 h of incubation. Aggrecan gene expression was increased in the presence of 0.1 ng/ml TNF-α. The fossa and disc cells responded to TNF-α with an increased expression of the aggrecanase ADAMTS4. TNF-α enhanced MMP-13 gene and protein expression only by condylar cells. Mechanical loading reduced this effect. Cells isolated from the different cartilaginous structures reacted differently to TNF-α. Since the disc and fossa contain a very low level of proteoglycans in comparison to the condyle, the role played by ADAMTS4 in degradation of the fossa and disc might be limited. TNF-α induced MMP-13 expression by condylar cells might be involved in the degradation of the juvenile condyle. Since this expression was reduced by mechanical loading, functional loading with oral physiotherapy or orthodontic activators may help to reduce the catabolic effect of TNF-α. J. Cell. Physiol. 232: 1287–1294, 2017. © 2016 Wiley Periodicals, Inc.Cells isolated from the different cartilaginous structures reacted differently to TNF-α. TNF-α enhanced MMP-13 gene and protein expression only by condylar cells. Mechanical loading reduced this effect.
      PubDate: 2017-01-06T10:30:23.595916-05:
      DOI: 10.1002/jcp.25593
       
  • Identifying Nuclear Matrix-Attached DNA Across the Genome
    • Authors: Jason R. Dobson; Deli Hong, A. Rasim Barutcu, Hai Wu, Anthony N. Imbalzano, Jane B. Lian, Janet L. Stein, Andre J. van Wijnen, Jeffrey A. Nickerson, Gary S. Stein
      Pages: 1295 - 1305
      Abstract: Experimental approaches to define the relationship between gene expression and nuclear matrix attachment regions (MARs) have given contrasting and method-specific results. We have developed a next generation sequencing strategy to identify MARs across the human genome (MAR-Seq). The method is based on crosslinking chromatin to its nuclear matrix attachment sites to minimize changes during biochemical processing. We used this method to compare nuclear matrix organization in MCF-10A mammary epithelial-like cells and MDA-MB-231 breast cancer cells and evaluated the results in the context of global gene expression (array analysis) and positional enrichment of gene-regulatory histone modifications (ChIP-Seq). In the normal-like cells, nuclear matrix-attached DNA was enriched in expressed genes, while in the breast cancer cells, it was enriched in non-expressed genes. In both cell lines, the chromatin modifications that mark transcriptional activation or repression were appropriately associated with gene expression. Using this new MAR-Seq approach, we provide the first genome-wide characterization of nuclear matrix attachment in mammalian cells and reveal that the nuclear matrix-associated genome is highly cell-context dependent. J. Cell. Physiol. 232: 1295–1305, 2017. © 2016 Wiley Periodicals, Inc.MAR-Seq is a next-generation sequencing strategy to identify matrix attachment regions (MARs) that is based on crosslinking chromatin to its nuclear attachment sites. Comparison of MCF-10A mammary epithelial-like cells and MDA-MB-231 breast cancer cells showed that nuclear matrix-attached DNA in normal-like MCF-10A cells is enriched in expressed genes, while in the breast cancer cells, it is enriched in non-expressed genes. This new MAR-Seq approach reveals that the nuclear matrix-associated genome is highly cell-context dependent.
      PubDate: 2017-01-05T08:35:24.082065-05:
      DOI: 10.1002/jcp.25596
       
  • A MicroRNA Cluster miR-23–24–27 Is Upregulated by Aldosterone in the
           Distal Kidney Nephron Where it Alters Sodium Transport
    • Authors: Xiaoning Liu; Robert S. Edinger, Christine A. Klemens, Yu L. Phua, Andrew J. Bodnar, William A. LaFramboise, Jacqueline Ho, Michael B. Butterworth
      Pages: 1306 - 1317
      Abstract: The epithelial sodium channel (ENaC) is expressed in the epithelial cells of the distal convoluted tubules, connecting tubules, and cortical collecting duct (CCD) in the kidney nephron. Under the regulation of the steroid hormone aldosterone, ENaC is a major determinant of sodium (Na+) and water balance. The ability of aldosterone to regulate microRNAs (miRs) in the kidney has recently been realized, but the role of miRs in Na+ regulation has not been well established. Here we demonstrate that expression of a miR cluster mmu-miR-23–24–27, is upregulated in the CCD by aldosterone stimulation both in vitro and in vivo. Increasing the expression of these miRs increased Na+ transport in the absence of aldosterone stimulation. Potential miR targets were evaluated and miR-27a/b was verified to bind to the 3′-untranslated region of intersectin-2, a multi-domain protein expressed in the distal kidney nephron and involved in the regulation of membrane trafficking. Expression of Itsn2 mRNA and protein was decreased after aldosterone stimulation. Depletion of Itsn2 expression, mimicking aldosterone regulation, increased ENaC-mediated Na+ transport, while Itsn2 overexpression reduced ENaC's function. These findings reinforce a role for miRs in aldosterone regulation of Na+ transport, and implicate miR-27 in aldosterone's action via a novel target. J. Cell. Physiol. 232: 1306–1317, 2017. © 2016 Wiley Periodicals, Inc.The steroid hormone aldosterone alters the expression of microRNAs in distal kidney nephron epithelial cells, and coordinately upregulates the miR-23–24–27 cluster. MiR-27 targets intersectin 2 to reduce its expression. Decreased intersectin 2 levels results in an increase in the activity of the epithelial sodium channel to augment sodium transport.
      PubDate: 2017-01-05T08:35:27.33984-05:0
      DOI: 10.1002/jcp.25599
       
  • Early Subchondral Bone Loss at Arthritis Onset Predicted Late Arthritis
           Severity in a Rat Arthritis Model
    • Authors: Guillaume Courbon; Damien Cleret, Marie-Thérèse Linossier, Laurence Vico, Hubert Marotte
      Pages: 1318 - 1325
      Abstract: Synovitis is usually observed before loss of articular function in rheumatoid arthritis (RA). In addition to the synovium and according to the “Inside–Outside” theory, bone compartment is also involved in RA pathogenesis. Then, we investigated time dependent articular bone loss and prediction of early bone loss to late arthritis severity on the rat adjuvant-induced arthritis (AIA) model. Lewis female rats were longitudinally monitored from arthritis induction (day 0), with early (day 10) and late (day 17) steps. Trabecular and cortical microarchitecture parameters of four ankle bones were assessed by microcomputed tomography. Gene expression was determined at sacrifice. Arthritis occurred at day 10 in AIA rats. At this time, bone erosions were detected on four ankle bones, with cortical porosity increase (+67%) and trabecular alterations including bone volume fraction (BV/TV: −13%), and trabecular thickness decrease. Navicular bone assessment was the most reproducible and sensitive. Furthermore, strong correlations were observed between bone alterations at day 10 and arthritis severity or bone loss at day 17, including predictability of day 10 BV/TV to day 17 articular index (R2 = 0.76). Finally, gene expression at day 17 confirmed massive osteoclast activation and interestingly provided insights on strong activation of bone formation inhibitor markers at the joint level. In rat AIA, bone loss was already observed at synovitis onset and was predicted late arthritis severity. Our results reinforced the key role of subchondral bone in arthritis pathogenesis, in favour to the “Inside–Outside” theory. Mechanisms of bone loss in rat AIA involved resorption activation and formation inhibition changes. J. Cell. Physiol. 232: 1318–1325, 2017. © 2016 Wiley Periodicals, Inc.We explored time dependent articular bone loss and prediction of early bone loss to late arthritis severity on the rat adjuvant-induced arthritis (AIA) model. At clinical synovitis onset, bone porosity and microarchitecture alterations were recorded in the ankle bones. These early modifications were predictive of arthritis outcome, underlining the importance of early bone loss investigation.
      PubDate: 2017-01-05T08:15:26.639024-05:
      DOI: 10.1002/jcp.25601
       
  • Mechanical Stress Regulates Bone Metabolism Through MicroRNAs
    • Authors: Yu Yuan; Lingli Zhang, Xiaoyang Tong, Miao Zhang, Yilong Zhao, Jianming Guo, Le Lei, Xi Chen, Jennifer Tickner, Jiake Xu, Jun Zou
      Pages: 1239 - 1245
      Abstract: There are many lines of evidence indicating that mechanical stress regulates bone metabolism and promotes bone growth. BMP, Wnt, ERK1/2, and OPG/RANKL are the main molecules thought to regulate the effects of mechanical loading on bone formation. Recently, microRNAs were found to be involved in bone cell proliferation and differentiation, regulating the balance of bone formation and bone resorption. Emerging evidence indicates that microRNAs also participate in mechanical stress-mediated bone metabolism, and is associated with disuse induced osteoporosis or osteopenia. Mechanical stress is able to induce expression of microRNAs that modulate the expression of osteogenic and bone resorption factors, leading to the positive impact of mechanical stress on bone. This review discusses the emerging evidence implicating an important role for microRNAs in the mechanical stress response in bone cells, as well as the challenges of translating microRNA research into potential treatment. J. Cell. Physiol. 232: 1239–1245, 2017. © 2016 Wiley Periodicals, Inc.Mechanical stress can induce expression of microRNAs that modulate the expression of osteogenic and bone resorption factors, leading to the positive impact of mechanical stress on bone. This review discusses the emerging evidence implicating an important role for microRNAs in the mechanical stress response in bone cells.
      PubDate: 2016-11-28T12:00:24.433488-05:
      DOI: 10.1002/jcp.25688
       
  • RNA-Generated and Gene-Edited Induced Pluripotent Stem Cells for Disease
           Modeling and Therapy
    • Authors: James Kehler; Marianna Greco, Valentina Martino, Manickam Pachiappan, Hiroko Yokoe, Alice Chen, Miranda Yang, Jonathan Auerbach, Joel Jessee, Martin Gotte, Luciano Milanesi, Alberto Albertini, Gianfranco Bellipanni, Ileana Zucchi, Rolland A. Reinbold, Antonio Giordano
      Pages: 1262 - 1269
      Abstract: Cellular reprogramming by epigenomic remodeling of chromatin holds great promise in the field of human regenerative medicine. As an example, human-induced Pluripotent Stem Cells (iPSCs) obtained by reprograming of patient somatic cells are sufficiently similar to embryonic stem cells (ESCs) and can generate all cell types of the human body. Clinical use of iPSCs is dependent on methods that do not utilize genome altering transgenic technologies that are potentially unsafe and ethically unacceptable. Transient delivery of exogenous RNA into cells provides a safer reprogramming system to transgenic approaches that rely on exogenous DNA or viral vectors. RNA reprogramming may prove to be more suitable for clinical applications and provide stable starting cell lines for gene-editing, isolation, and characterization of patient iPSC lines. The introduction and rapid evolution of CRISPR/Cas9 gene-editing systems has provided a readily accessible research tool to perform functional human genetic experiments. Similar to RNA reprogramming, transient delivery of mRNA encoding Cas9 in combination with guide RNA sequences to target specific points in the genome eliminates the risk of potential integration of Cas9 plasmid constructs. We present optimized RNA-based laboratory procedure for making and editing iPSCs. In the near-term these two powerful technologies are being harnessed to dissect mechanisms of human development and disease in vitro, supporting both basic, and translational research. J. Cell. Physiol. 232: 1262–1269, 2017. © 2016 Wiley Periodicals, Inc.Transient delivery of exogenous RNA into cells provides a safer reprogramming system to transgenic approaches that rely on exogenous DNA or viral vectors. RNA reprogrammed iPSCs lines may prove to be more suitable for clinical applications and provide stable starting cell lines for gene-editing, isolation, and characterization of patient iPSC lines. The introduction and rapid evolution of CRISPR/Cas9 gene-editing systems has provided a readily accessible research tool to perform functional human genetic experiments. Similar to RNA reprogramming, transient delivery of mRNA encoding Cas9 in combination with guide RNA sequences to target specific points in the genome eliminates the risk of potential integration of Cas9 plasmid constructs. We present optimized RNA-based laboratory procedures for making and editing iPSCs. These two powerful technologies are being harnessed to dissect mechanisms of human development and disease in vitro, supporting both basic and translational research.
      PubDate: 2016-12-20T08:30:50.328732-05:
      DOI: 10.1002/jcp.25597
       
  • Exploring the Role of PGC-1α in Defining Nuclear Organisation in
           Skeletal Muscle Fibres
    • Authors: Jacob Alexander Ross; Adam Pearson, Yotam Levy, Bettina Cardel, Christoph Handschin, Julien Ochala
      Pages: 1270 - 1274
      Abstract: Muscle fibres are multinucleated cells, with each nucleus controlling the protein synthesis in a finite volume of cytoplasm termed the myonuclear domain (MND). What determines MND size remains unclear. In the present study, we aimed to test the hypothesis that the level of expression of the transcriptional coactivator PGC-1α and subsequent activation of the mitochondrial biogenesis are major contributors. Hence, we used two transgenic mouse models with varying expression of PGC-1α in skeletal muscles. We isolated myofibres from the fast twitch extensor digitorum longus (EDL) and slow twitch diaphragm muscles. We then membrane-permeabilised them and analysed the 3D spatial arrangements of myonuclei. In EDL muscles, when PGC-1α is over-expressed, MND volume decreases; whereas, when PGC-1α is lacking, no change occurs. In the diaphragm, no clear difference was noted. This indicates that PGC-1α and the related mitochondrial biogenesis programme are determinants of MND size. PGC-1α may facilitate the addition of new myonuclei in order to reach MND volumes that can support an increased mitochondrial density. J. Cell. Physiol. 232: 1270–1274, 2017. © 2016 Wiley Periodicals, Inc.Muscle fibres are multinucleated cells, with individual nuclei controlling the protein synthesis in a finite volume of cytoplasm termed myonuclear domain. The present study indicates that one important regulator of myonuclear domain size is PGC-1α and related mitochondrial biogenesis programme.
      PubDate: 2016-12-29T05:37:05.80091-05:0
      DOI: 10.1002/jcp.25678
       
  • Acute Ethanol Increases IGF-I-Induced Phosphorylation of ERKs by Enhancing
           Recruitment of p52-Shc to the Grb2/Shc Complex
    • Authors: Matthew Dean; Adam Lassak, Anna Wilk, Adriana Zapata, Luis Marrero, Patricia Molina, Krzysztof Reiss
      Pages: 1275 - 1286
      Abstract: Ethanol plays a detrimental role in the development of the brain. Multiple studies have shown that ethanol inhibits insulin-like growth factor I receptor (IGF-IR) function. Because the IGF-IR contributes to brain development by supporting neural growth, survival, and differentiation, we sought to determine the molecular mechanism(s) involved in ethanol's effects on this membrane-associated tyrosine kinase. Using multiple neuronal cell types, we performed Western blot, immunoprecipitation, and GST-pulldowns following acute (1–24 h) or chronic (3 weeks) treatment with ethanol. Surprisingly, exposure of multiple neuronal cell types to acute (up to 24 h) ethanol (50 mM) enhanced IGF-I-induced phosphorylation of extracellular regulated kinases (ERKs), without affecting IGF-IR tyrosine phosphorylation itself, or Akt phosphorylation. This acute increase in ERKs phosphorylation was followed by the expected inhibition of the IGF-IR signaling following 3-week ethanol exposure. We then expressed a GFP-tagged IGF-IR construct in PC12 cells and used them to perform fluorescence recovery after photobleaching (FRAP) analysis. Using these fluorescently labeled cells, we determined that 50 mM ethanol decreased the half-time of the IGF-IR-associated FRAP, which implied that cell membrane-associated signaling events could be affected. Indeed, co-immunoprecipitation and GST-pulldown studies demonstrated that the acute ethanol exposure increased the recruitment of p52-Shc to the Grb2-Shc complex, which is known to engage the Ras-Raf-ERKs pathway following IGF-1 stimulation. These experiments indicate that even a short and low-dose exposure to ethanol may dysregulate function of the receptor, which plays a critical role in brain development. J. Cell. Physiol. 232: 1275–1286, 2017. © 2016 Wiley Periodicals, Inc.In the current study, we show a novel effect of ethanol on the IGF-I receptor (IGF-IR) signaling, and provided a possible mechanism through which this occurs. We determine that ethanol enhances mobility of the IGF-IR molecules within the cell membrane, and that this stimulatory effect enhances formation of the Grb2-Shc signaling complex leading to the transient increase of the signal toward extracellular regulated kinases (ERKs). Because of the critical role of the IGF-IR signaling in neuronal growth and survival, and the detrimental effects of alcohol on CNS development, we believe that our new data will help in better understanding of the fetal alcohol syndrome.
      PubDate: 2016-12-19T11:50:27.245701-05:
      DOI: 10.1002/jcp.25586
       
  • Differential Expression of miR-4520a Associated With Pyrin Mutations in
           Familial Mediterranean Fever (FMF)
    • Authors: Helen Latsoudis; Mir-Farzin Mashreghi, Joachim R. Grün, Hyun-Dong Chang, Bruno Stuhlmüller, Argyro Repa, Irini Gergiannaki, Eleni Kabouraki, George S. Vlachos, Thomas Häupl, Andreas Radbruch, Prodromos Sidiropoulos, Kimon Doukoumetzidis, Dimitris Kardassis, Timothy B. Niewold, Dimitrios T. Boumpas, George N. Goulielmos
      Pages: 1326 - 1336
      Abstract: Familial Mediterranean fever (FMF) is an autosomal recessive disease characterized by recurrent, acute, and self-limiting attacks of fever. Mutations in MEFV gene encoding pyrin account for FMF, but the high number of heterozygote patients with typical symptoms of the disease has driven a number of alternative aetiopathogenic hypotheses. The MEFV gene was knocked down in human myelomonocytic cells that express endogenous pyrin to identify deregulated microRNAs (miRNAs). Microarray analyses revealed 29 significantly differentially expressed miRNAs implicated in pathways associated with cellular integrity and survival. Implementation of in silico gene network prediction algorithms and bioinformatics analyses showed that miR-4520a is predicted to target genes implicated in autophagy through regulation of RHEB/mTOR signaling. Differential expression levels of RHEB were confirmed by luciferase reporter gene assays providing further evidence that is directly targeted by miR-4520a. Although the relative expression levels of miR-4520a were variable among FMF patients, the statistical expression of miR-4520a was different between FMF mutation carriers and controls (P = 0.0061), indicating an association between miR-4520a expression and MEFV mutations. Comparison between FMF patients bearing the M694V mutation, associated with severe disease, and healthy controls showed a significant increase in miR-4520a expression levels (P = 0.00545). These data suggest that RHEB, the main activator of mTOR signaling, is a valid target of miR-4520a with the relative expression levels of the latter being significantly deregulated in FMF patients and highly dependent on the presence of pyrin mutations, especially of the M694V type. These results suggest a role of deregulated autophagy in the pathogenesis of FMF. J. Cell. Physiol. 232: 1326–1336, 2017. © 2016 Wiley Periodicals, Inc.Familial Mediterranean fever (FMF) is an autosomal recessive disease characterized by recurrent, acute and self-limiting attacks of fever. In the present study we presented for the first time results supporting a role of deregulated autophagy in the pathogenesis of FMF.
      PubDate: 2016-12-20T08:30:57.563042-05:
      DOI: 10.1002/jcp.25602
       
  • MED28 Regulates Epithelial–Mesenchymal Transition Through NFκB in Human
           Breast Cancer Cells
    • Authors: Chun-Yin Huang; Nien-Tsu Hsieh, Chun-I Li, Yu-Ting Weng, Hsiao-Sheng Liu, Ming-Fen Lee
      Pages: 1337 - 1345
      Abstract: MED28, a mammalian Mediator subunit, was found highly expressed in several types of malignancy, including breast cancer. Recently, we have identified a role of MED28 in regulating both cell growth and migration in human breast cancer cells. In epithelium-derived solid tumor, migration and invasion are preceded by the progression of epithelial–mesenchymal transition (EMT) which calls for downregulation of epithelial markers as well as upregulation of mesenchymal markers, among other features. The objective of this study was to investigate a putative role of MED28 in the progression of EMT in human breast cancer cells. In fibroblast-like MDA-MB-231 cells, suppression of MED28 attenuated the mesenchymal morphology, concomitantly with a reduction of several mesenchymal biomarkers and Snail, a transcriptional repressor of E-cadherin. The suppression effect was also accompanied by downregulation of p-NFκB/p65. However, overexpression of MED28 exhibited in an opposite manner. In epithelial MCF7 cells, administration of Adriamycin®, an experimental EMT induction system, led to a mesenchyme-like appearance correlated with increased expression of MED28, p-p65, and Snail, and a reciprocal change of epithelial and mesenchymal markers. Furthermore, suppression of MED28 attenuated the experimental EMT effect and restored the original expression status of E-cadherin and MMP9 in MCF7 cells. Our data indicate that MED28 modulates the development of EMT through NFκB in human breast cancer cells, further reinforcing the significance of MED28 in the progression of breast cancer on top of its role in cell growth and migration. J. Cell. Physiol. 232: 1337–1345, 2017. © 2016 Wiley Periodicals, Inc.MED28 regulates the expression of epithelial markers and mesenchymal markers through NFκB, thereby modulating the development of epithelial–mesenchymal transition. These findings further reinforce the significance of MED28 in the progression of breast cancer on top of its role in cell growth and migration.
      PubDate: 2016-09-30T09:09:02.969848-05:
      DOI: 10.1002/jcp.25610
       
  • Hypoglycemia Enhances Epithelial-Mesenchymal Transition and Invasiveness,
           and Restrains the Warburg Phenotype, in Hypoxic HeLa Cell Cultures and
           Microspheroids
    • Authors: Álvaro Marín-Hernández; Juan Carlos Gallardo-Pérez, Ileana Hernández-Reséndiz, Isis Del Mazo-Monsalvo, Diana Xochiquetzal Robledo-Cadena, Rafael Moreno-Sánchez, Sara Rodríguez-Enríquez
      Pages: 1346 - 1359
      Abstract: The accelerated growth of solid tumors leads to episodes of both hypoxia and hypoglycemia (HH) affecting their intermediary metabolism, signal transduction, and transcriptional activity. A previous study showed that normoxia (20% O2) plus 24 h hypoglycemia (2.5 mM glucose) increased glycolytic flux whereas oxidative phosphorylation (OxPhos) was unchanged versus normoglycemia in HeLa cells. However, the simultaneous effect of HH on energy metabolism has not been yet examined. Therefore, the effect of hypoxia (0.1–1% O2) plus hypoglycemia on the energy metabolism of HeLa cells was analyzed by evaluating protein content and activity, along with fluxes of both glycolysis and OxPhos. Under hypoxia, in which cell growth ceased and OxPhos enzyme activities, ΔΨm and flux were depressed, hypoglycemia did not stimulate glycolytic flux despite increasing H-RAS, p-AMPK, GLUT1, GLUT3, and HKI levels, and further decreasing mitochondrial enzyme content. The impaired mitochondrial function in HH cells correlated with mitophagy activation. The depressed OxPhos and unchanged glycolysis pattern was also observed in quiescent cells from mature multicellular tumor spheroids, suggesting that these inner cell layers are similarly subjected to HH. The principal ATP supplier was glycolysis for HH 2D monolayer and 3D quiescent spheroid cells. Accordingly, the glycolytic inhibitors iodoacetate and gossypol were more effective than mitochondrial inhibitors in decreasing HH-cancer cell viability. Under HH, stem cell-, angiogenic-, and EMT-biomarkers, as well as glycoprotein-P content and invasiveness, were also enhanced. These observations indicate that HH cancer cells develop an attenuated Warburg and pronounced EMT- and invasive-phenotype. J. Cell. Physiol. 232: 1346–1359, 2017. © 2016 Wiley Periodicals, Inc.Hypoglycemia enhances (i) epithelial-mesenchymal transition, (ii) invasiveness, and (iii) restrains the Warburg phenotype in hypoxic HeLa cells.
      PubDate: 2016-09-30T09:16:31.539215-05:
      DOI: 10.1002/jcp.25617
       
  • p120-Catenin Is Required for Dietary Calcium Suppression of Oral
           Carcinogenesis in Mice
    • Authors: Zhongjian Xie; Yuan Yuan, Yi Jiang, Chandrama Shrestha, Ying Chen, Liyan Liao, Shangli Ji, Xiaoge Deng, Eryuan Liao, Daniel D. Bikle
      Pages: 1360 - 1367
      Abstract: Previous studies have shown that dietary calcium suppresses oral carcinogenesis, but the mechanism is unclear. p120-catenin (p120) is a cytoplasmic protein closely associated with E-cadherin to form the E-cadherin–β-catenin complex and may function as a tumor suppressor in the oral epithelium. To determine whether p120 is involved in the mechanism by which dietary calcium suppresses oral carcinogenesis, The normal, low, or high calcium diet was fed control mice (designated as floxed p120 mice) or mice in which p120 was specifically deleted in the oral squamous epithelium during the adult stage (designated as p120cKO mice). All mice were exposed to a low dose of oral cancer carcinogen 4-nitroquinoline 1-oxide and rates of oral squamous cell carcinoma (OSCC) and proliferation and differentiation in the cancerous and non-cancerous oral epithelium of these mice were examined. The results showed that the low calcium diet increased rates of OSCC and proliferation of the non-cancerous oral epithelium and decreased differentiation of the non-cancerous oral epithelium, but had no effect on cancerous oral epithelium. In contrast, the high calcium diet had opposite effects. However, the effect of the dietary calcium on the rates of OSCC, proliferation, and differentiation of the non-cancerous epithelium were not seen in p120cKO mice. Based on these results, we conclude that p120 is required for dietary calcium suppression of oral carcinogenesis and oral epithelial proliferation and dietary calcium induction of oral epithelial differentiation. J. Cell. Physiol. 232: 1360–1367, 2017. © 2016 Wiley Periodicals, Inc.The aim of the present study was to determine whether p120-catenin plays a role in dietary calcium-induced oral carcinogenesis. The results showed that knockout of p120-catenin blocked the inhibitory effect of dietary calcium on tumor rates and proliferation and the stimulatory effect of dietary calcium on tumor differentiation in the oral epithelium. These data suggest that p120-catenin plays a critical role in dietary calcium-suppressed carcinogenesis and dietary calcium regulation of proliferation, differentiation. Our findings illustrate a mechanism by which dietary calcium suppresses oral carcinogenesis will help in the development of better preventive and therapeutic strategies for oral cancers.
      PubDate: 2016-11-10T06:40:29.912919-05:
      DOI: 10.1002/jcp.25620
       
  • Identification of a Novel Human E-Cadherin Splice Variant and Assessment
           of Its Effects Upon EMT-Related Events
    • Authors: María Laura Matos; Lara Lapyckyj, Marina Rosso, María José Besso, María Victoria Mencucci, Clara Isabel Marín Briggiler, Silvina Giustina, Laura Inés Furlong, Mónica Hebe Vazquez-Levin
      Pages: 1368 - 1386
      Abstract: Epithelial Cadherin (E-cadherin) is involved in calcium-dependent cell–cell adhesion and signal transduction. The E-cadherin decrease/loss is a hallmark of Epithelial to Mesenchymal Transition (EMT), a key event in tumor progression. The underlying molecular mechanisms that trigger E-cadherin loss and consequent EMT have not been completely elucidated. This study reports the identification of a novel human E-cadherin variant mRNA produced by alternative splicing. A bioinformatics evaluation of the novel mRNA sequence and biochemical verifications suggest its regulation by Nonsense-Mediated mRNA Decay (NMD). The novel E-cadherin variant was detected in 29/42 (69%) human tumor cell lines, expressed at variable levels (E-cadherin variant expression relative to the wild type mRNA = 0.05–11.6%). Stable transfection of the novel E-cadherin variant in MCF-7 cells (MCF7Ecadvar) resulted in downregulation of wild type E-cadherin expression (transcript/protein) and EMT-related changes, among them acquisition of a fibroblastic-like cell phenotype, increased expression of Twist, Snail, Zeb1, and Slug transcriptional repressors and decreased expression of ESRP1 and ESRP2 RNA binding proteins. Moreover, loss of cytokeratins and gain of vimentin, N-cadherin and Dysadherin/FXYD5 proteins was observed. Dramatic changes in cell behavior were found in MCF7Ecadvar, as judged by the decreased cell–cell adhesion (Hanging-drop assay), increased cell motility (Wound Healing) and increased cell migration (Transwell) and invasion (Transwell w/Matrigel). Some changes were found in MCF-7 cells incubated with culture medium supplemented with conditioned medium from HEK-293 cells transfected with the E-cadherin variant mRNA. Further characterization of the novel E-cadherin variant will help understanding the molecular basis of tumor progression and improve cancer diagnosis. J. Cell. Physiol. 232: 1368–1386, 2017. © 2016 Wiley Periodicals, Inc.This report describes the identification and partial characterization of a novel human E-cadherin variant mRNA produced by alternative splicing. Stable transfection of the E-cadherin variant mRNA in MCF-7 cells (MCF7Ecadvar cells) resulted in downregulation of wild type E-cadherin and EMT-related changes, that is, acquisition of a fibroblast-like cellular phenotype, increased expression of E-cadherin transcriptional repressors, vimentin and N-cadherin, and loss of cytokeratins. MCF7Ecadvar cells depicted dramatic changes in cellular behavior, showing reduced cell–cell adhesion and increased cell migration and invasiveness.
      PubDate: 2016-12-29T05:36:52.820603-05:
      DOI: 10.1002/jcp.25622
       
  • MicroRNA-494 Activation Suppresses Bone Marrow Stromal Cell-Mediated Drug
           Resistance in Acute Myeloid Leukemia Cells
    • Authors: Chen Tian; Guoguang Zheng, Hongqing Zhuang, Xubin Li, Dongzhi Hu, Lei Zhu, Tengteng Wang, Mingjian James You, Yizhuo Zhang
      Pages: 1387 - 1395
      Abstract: Acute myeloid leukemia (AML) is not sensitive to chemotherapy partially because of the protection of AML cells by mesenchymal stromal cells (MSCs). Our previous studies found that MSCs protected AML cells from apoptosis through the c-Myc-dependent pathway. However, the mechanism by which MSCs regulate c-Myc in AML cells is still unknown. To elucidate the mechanism, we performed microRNA array analysis of AML cell lines and validated by TaqMan realtime PCR. The results showed that the expression of microRNA-494 (miR-494) in AML cells after coculture with MSCs was downregulated. Reporter gene analysis confirmed miR-494 as one of the regulators of c-Myc. In the coculture system, activation of miR-494 in AML cells suppressed proliferation and induced apoptosis of AML cells in vitro. After addition of mitoxantrone to the coculture system, the proliferation of AML cells with miR-494 activation was suppressed more than that of control cells. After subcutaneous injection of AML cell lines in combination with MSC, tumor growth was suppressed in mice injected with miR-494-overexpressing AML cells. The rate of tumor formation was even lower after mitoxantrone treatment in the miR-494 overexpressing group. Moreover, miR-494 activation resulted in a decrease of leukemic cell counts in peripheral blood (PB) and bone marrow, and prolonged survival in mice injected with miR-494-overexpressing AML cellls and MSCs compared to the control mice. Our results indicate that miR-494 suppresses drug resistance in AML cells by downregulating c-Myc through interaction with MSCs and that miR-494 therefore is a potential therapeutic target. J. Cell. Physiol. 232: 1387–1395, 2017. © 2016 Wiley Periodicals, Inc.miR-494 suppresses drug resistance in AML cells by downregulating c-Myc through interaction with MSCs and that miR-494 therefore is a potential therapeutic target.
      PubDate: 2016-10-19T11:25:27.872055-05:
      DOI: 10.1002/jcp.25628
       
  • Osteoclast Fusion: Time-Lapse Reveals Involvement of CD47 and Syncytin-1
           at Different Stages of Nuclearity
    • Authors: Anaïs Marie Julie Møller; Jean-Marie Delaissé, Kent Søe
      Pages: 1396 - 1403
      Abstract: Investigations addressing the molecular keys of osteoclast fusion are primarily based on end-point analyses. No matter if investigations are performed in vivo or in vitro the impact of a given factor is predominantly analyzed by counting the number of multi-nucleated cells, the number of nuclei per multinucleated cell or TRAcP activity. But end-point analyses do not show how the fusion came about. This would not be a problem if fusion of osteoclasts was a random process and occurred by the same molecular mechanism from beginning to end. However, we and others have in the recent period published data suggesting that fusion partners may specifically select each other and that heterogeneity between the partners seems to play a role. Therefore, we set out to directly test the hypothesis that fusion factors have a heterogenic involvement at different stages of nuclearity. Therefore, we have analyzed individual fusion events using time-lapse and antagonists of CD47 and syncytin-1. All time-lapse recordings have been studied by two independent observers. A total of 1808 fusion events were analyzed. The present study shows that CD47 and syncytin-1 have different roles in osteoclast fusion depending on the nuclearity of fusion partners. While CD47 promotes cell fusions involving mono-nucleated pre-osteoclasts, syncytin-1 promotes fusion of two multi-nucleated osteoclasts, but also reduces the number of fusions between mono-nucleated pre-osteoclasts. Furthermore, CD47 seems to mediate fusion mostly through broad contact surfaces between the partners’ cell membrane while syncytin-1 mediate fusion through phagocytic-cup like structure. J. Cell. Physiol. 232: 1396–1403, 2017. © 2016 Wiley Periodicals, Inc.Time-lapse recording has allowed us to categorize cell fusion events according to the number of nuclei of the fusion partners. We show here that factors commonly believed to be critical for fusion, are in fact only involved in some categories of fusion. This points to the existence of distinct molecular mechanisms of cell fusion depending on the degree of nuclearity.
      PubDate: 2016-10-19T11:20:25.747768-05:
      DOI: 10.1002/jcp.25633
       
  • Essential Role of CFTR in PKA-Dependent Phosphorylation, Alkalinization,
           and Hyperpolarization During Human Sperm Capacitation
    • Authors: Lis C. Puga Molina; Nicolás A. Pinto, Paulina Torres Rodríguez, Ana Romarowski, Alberto Vicens Sanchez, Pablo E. Visconti, Alberto Darszon, Claudia L. Treviño, Mariano G. Buffone
      Pages: 1404 - 1414
      Abstract: Mammalian sperm require to spend a limited period of time in the female reproductive tract to become competent to fertilize in a process called capacitation. It is well established that HCO3− is essential for capacitation because it activates the atypical soluble adenylate cyclase ADCY10 leading to cAMP production, and promotes alkalinization of cytoplasm, and membrane hyperpolarization. However, how HCO3− is transported into the sperm is not well understood. There is evidence that CFTR activity is involved in the human sperm capacitation but how this channel is integrated in the complex signaling cascades associated with this process remains largely unknown. In the present work, we have analyzed the extent to which CFTR regulates different events in human sperm capacitation. We observed that inhibition of CFTR affects HCO3−-entrance dependent events resulting in lower PKA activity. CFTR inhibition also affected cAMP/PKA-downstream events such as the increase in tyrosine phosphorylation, hyperactivated motility, and acrosome reaction. In addition, we demonstrated for the first time, that CFTR and PKA activity are essential for the regulation of intracellular pH, and membrane potential in human sperm. Addition of permeable cAMP partially recovered all the PKA-dependent events altered in the presence of inh-172 which is consistent with a role of CFTR upstream of PKA activation. J. Cell. Physiol. 232: 1404–1414, 2017. © 2016 Wiley Periodicals, Inc.We observed that inhibition of CFTR affects HCO3-entrance dependent events resulting in lower PKA activity.
      PubDate: 2016-10-26T09:05:27.52414-05:0
      DOI: 10.1002/jcp.25634
       
  • Short- and Long-Term Hindlimb Immobilization and Reloading: Profile of
           Epigenetic Events in Gastrocnemius
    • Authors: Alba Chacon-Cabrera; Joaquim Gea, Esther Barreiro
      Pages: 1415 - 1427
      Abstract: Skeletal muscle dysfunction and atrophy are characteristic features accompanying chronic conditions. Epigenetic events regulate muscle mass and function maintenance. We hypothesized that the pattern of epigenetic events (muscle-enriched microRNAs and histone acetylation) and acetylation of transcription factors known to signal muscle wasting may differ between early- and late-time points in skeletal muscles of mice exposed to hindlimb immobilization (I) and recovery following I. Body and muscle weights, grip strength, muscle-enriched microRNAs, histone deacetylases (HDACs), acetylation of proteins, histones, and transcription factors (TF), myogenic TF factors, and muscle phenotype were assessed in gastrocnemius of mice exposed to periods (1, 2, 3, 7, 15, and 30 days, I groups) of hindlimb immobilization, and in those exposed to reloading for different periods of time (1, 3, 7, 15, and 30 days, R groups) following 7-day immobilization. Compared to non-immobilized controls, muscle weight, limb strength, microRNAs, especially miR-486, SIRT1 levels, and slow- and fast-twitch cross-sectional areas were decreased in mice of I groups, whereas Pax7 and acetylated FoxO1 and FoxO3 levels were increased. Muscle reloading following splint removal improved muscle mass loss, strength, and fiber atrophy, by increasing microRNAs, particularly miR-486, and SIRT1 content, while decreasing acetylated FoxO1 and FoxO3 levels. In this mouse model of disuse muscle atrophy, muscle-enriched microRNAs, especially miR-486, through Pax7 regulation delayed muscle cell differentiation following unloading of gastrocnemius muscle. Acetylation of FoxO1 and 3 seemed to drive muscle mass loss and atrophy, while deacetylation of these factors through SIRT1 would enable the muscle fibers to regenerate. J. Cell. Physiol. 232: 1415–1427, 2017. © 2016 Wiley Periodicals, Inc.In this mouse model of disuse muscle atrophy, muscle-enriched microRNAs, especially miR-486, through Pax7 regulation delayed muscle cell differentiation following unloading of gastrocnemius muscle. Acetylation of FoxO1 and 3 seemed to drive muscle mass loss and atrophy, while deacetylation of these factors through SIRT1 would enable the muscle fibers to regenerate.
      PubDate: 2016-10-19T11:25:25.182241-05:
      DOI: 10.1002/jcp.25635
       
  • Inhibitory Effects of Quercetin on Progression of Human Choriocarcinoma
           
    • Authors: Whasun Lim; Changwon Yang, Sunwoo Park, Fuller W. Bazer, Gwonhwa Song
      Pages: 1428 - 1440
      Abstract: As a major dietary flavonol, quercetin mitigates proliferation and progression of cancer due to its anti-angiogenic, anti-inflammatory, anti-oxidant, and apoptotic biological effects on cells. Although its apoptotic effects have been reported for various cancers, little is known of the functional role of quercetin in gestational choriocarcinoma. Results of the present study indicated that quercetin reduced proliferation and induced cell death in two choriocarcinoma cell lines, JAR and JEG3 cells, with an increase in the sub-G1 phase of the cell cycle. In addition, quercetin induced mitochondrial dysfunction significantly reduced mitochondrial membrane potential (MMP) and increased production of reactive oxygen species (ROS) in both JAR and JEG3 cells. Further, quercetin inhibited phosphorylation of AKT, P70S6K and S6 proteins whereas, it increased phosphorylation of ERK1/2, P38, JNK and P90RSK proteins in JAR and JEG3 cells. The decrease in viability of choriocarcinoma cells treated with quercetin was confirmed by using combinations of quercetin and pharmacological inhibitors of the PI3K and MAPK signaling pathways. Classical chemotherapeutic agents, cisplatin (a platinum-based drug) and paclitaxel (a taxene-based drug), inhibited proliferation of JAR and JEG3 cells, and when combined with quercetin, the antiproliferative effects of cisplatin and paclitaxel were enhanced for both choriocarcinoma cell lines. Collectively, these results suggest that quercetin prevents development of choriocarcinoma and may be a valuable therapeutic agent for treatment of choriocarcinoma through its regulation of PI3K and MAPK signal transduction pathways. J. Cell. Physiol. 232: 1428–1440, 2017. © 2016 Wiley Periodicals, Inc.Quercetin prevents development of choriocarcinoma and may be a valuable therapeutic agent for treatment of choriocarcinoma through its regulation of PI3K/AKT and MAPK signaling pathways and intrinsic apoptotic pathways.
      PubDate: 2016-11-30T10:21:09.529104-05:
      DOI: 10.1002/jcp.25637
       
  • Peripheral Blood Mononuclear Cells Infiltration Downregulates Decidual
           FAAH Activity in an LPS-Induced Embryo Resorption Model
    • Authors: Manuel Luis Wolfson; Julieta Aisemberg, Fernando Correa, Ana María Franchi
      Pages: 1441 - 1447
      Abstract: Maternal infections with gram-negative bacteria are associated with miscarriage and are one of the most common complications during pregnancy. Previous studies from our group have shown that lipopolysaccharide (LPS)-activated infiltrating peripheral blood mononuclear cells (PBMC) into decidual tissue plays an important role in the establishment of a local inflammatory process that results in embryo cytotoxicity and early embryo resorption. Moreover, we have also shown that an increased endocannabinoid tone mediates LPS-induced deleterious effects during early pregnancy loss. Here, we sought to investigate whether the infiltrating PBMC modulates the decidual endocannabinoid tone and the molecular mechanisms involved. PBMC isolated from 7-day pregnant mice subjected to different treatments were co-cultured in a transwell system with decidual tissue from control 7-day pregnant mice. Decidual fatty acid amide hydrolase (FAAH) activity was measured by radioconvertion, total decidual protein nitration by Western blot (WB), and decidual FAAH nitration by immunoprecipitation followed by WB. We found that co-culture of PBMC obtained from LPS-treated mice increased the level of nitration of decidual FAAH, which resulted in a negative modulation of decidual FAAH activity. Interestingly, co-treatment with progesterone or aminoguanidine prevented this effect. We found that LPS-treated PBMC release high amounts of nitric oxide (NO) which causes tyrosine nitration of decidual FAAH, diminishing its enzymatic activity. Inactivation of FAAH, the main degrading enzyme of anandamide and similar endocannabinoids, could lead to an increased decidual endocannabinoid tone with embryotoxic effects. J. Cell. Physiol. 232: 1441–1447, 2017. © 2016 Wiley Periodicals, Inc.PBMC release a soluble factor which modulates the decidual FAAH activity. Peroxynitrite participates in LPS-treated PBMC decidual FAAH activity modulation. PBMC from LPS-treated mice increased the nitration levels of decidual FAAH which resulted in a decreased enzymatic activity.
      PubDate: 2016-11-10T06:51:10.223851-05:
      DOI: 10.1002/jcp.25640
       
  • Low Shear Stress Attenuates COX-2 Expression Induced by Resistin in Human
           Osteoarthritic Chondrocytes
    • Authors: Yu-Ping Su; Cheng-Nan Chen, Hsin-I Chang, Kuo-Chin Huang, Chin-Chang Cheng, Fang-Yao Chiu, Ko-Chao Lee, Chun-Min Lo, Shun-Fu Chang
      Pages: 1448 - 1457
      Abstract: Low shear stress has been proposed to play a reparative role in modulating cartilage homeostasis. Recently, epidemiological studies have found a positive correlation between the resistin level in serum and synovial fluid and osteoarthritis (OA) severity in patients. However, the effect of moderate shear stress on the catabolic stimulation of resistin in OA chondrocytes remains unclear. Hence, this study was to investigate whether low shear stress could regulate resistin-induced catabolic cyclooxygenase (COX)-2 expression in human OA chondrocytes and the underlying mechanism. Human OA chondrocytes and SW1353 chondrosarcoma cells were used in this study. Two modes of low shear stress (2 dyn/cm2), pre-shear and post-shear, were applied to the chondrocytes. A specific activator and siRNAs were used to investigate the mechanism of low shear stress-regulated COX-2 expression of resistin induction. We found that human OA chondrocytes exposed to different modes of low shear stress elicit an opposite effect on resistin-induced COX-2 expression: pre-shear for a short duration attenuates the resistin effect by inhibiting the transcription factor nuclear factor (NF)-κB-p65 subunit and the cAMP response element binding protein; however, post-shear over a longer duration enhances the resistin effect by activating only the NF-κB-p65 subunit. Moreover, our results demonstrated that the regulation of both shear modes in resistin-stimulated COX-2 expression occurs through increasing AMP-activated protein kinase activation and then sirtuin 1 expression. This study elucidates the detailed mechanism of low shear stress regulating the resistin-induced catabolic COX-2 expression and indicates a possible reparative role of moderate shear force in resistin-stimulated OA development. J. Cell. Physiol. 232: 1448–1457, 2017. © 2016 Wiley Periodicals, Inc.This study elucidates the detailed mechanism of low shear stress regulating the resistin-induced catabolic COX-2 expression and indicates a possible reparative role of moderate shear force in resistin-stimulated OA development.
      PubDate: 2016-11-20T15:45:39.505378-05:
      DOI: 10.1002/jcp.25644
       
  • PPARα Antagonist AA452 Triggers Metabolic Reprogramming and Increases
           Sensitivity to Radiation Therapy in Human Glioblastoma Primary Cells
    • Authors: Elisabetta Benedetti; Michele d'Angelo, Alessandra Ammazzalorso, Giovanni Luca Gravina, Chiara Laezza, Andrea Antonosante, Gloria Panella, Benedetta Cinque, Loredana Cristiano, Anne Chloè Dhez, Carlo Astarita, Renato Galzio, Maria Grazia Cifone, Rodolfo Ippoliti, Rosa Amoroso, Ernesto Di Cesare, Antonio Giordano, Annamaria Cimini
      Pages: 1458 - 1466
      Abstract: Glioblastoma (GB) is the most common cancer in the brain and with an increasing incidence. Despite major advances in the field, there is no curative therapy for GB to date. Many solid tumors, including GB, experienced metabolic reprogramming in order to sustain uncontrolled proliferation, hypoxic conditions, and angiogenesis. PPARs, member of the steroid hormone receptor superfamily, are particularly involved in the control of energetic metabolism, particularly lipid metabolism, which has been reported deregulated in gliomas. PPARα was previously indicated by us as a potential therapeutic target for this neoplasm, due to the malignancy grade dependency of its expression, being particularly abundant in GB. In this work, we used a new PPARα antagonist on patient-derived GB primary cells, with particular focus on the effects on lipid metabolism and response to radiotherapy. The results obtained demonstrated that blocking PPARα results in cell death induction, increase of radiosensitivity, and decrease of migration. Therefore, AA452 is proposed as a new adjuvant for the gold standard therapies for GB, opening the possibility for preclinical and clinical trials for this class of compounds. J. Cell. Physiol. 232: 1458–1466, 2017. © 2016 Wiley Periodicals, Inc.A new compound, an inhibitor of PPARalpha trancriptional activity, is reported, able to increase radiation sensitivity of glioblastoma cells.
      PubDate: 2016-11-30T10:16:56.191194-05:
      DOI: 10.1002/jcp.25648
       
  • Primary Cilium-Regulated EG-VEGF Signaling Facilitates Trophoblast
           Invasion
    • Authors: Chia-Yih Wang; Hui-Ling Tsai, Jhih-Siang Syu, Ting-Yu Chen, Mei-Tsz Su
      Pages: 1467 - 1477
      Abstract: Trophoblast invasion is an important event in embryo implantation and placental development. During these processes, endocrine gland-derived vascular endothelial growth factor (EG-VEGF) is the key regulator mediating the crosstalk at the feto-maternal interface. The primary cilium is a cellular antenna receiving environmental signals and is crucial for proper development. However, little is known regarding the role of the primary cilium in early human pregnancy. Here, we demonstrate that EG-VEGF regulates trophoblast cell invasion via primary cilia. We found that EG-VEGF activated ERK1/2 signaling and subsequent upregulation of MMP2 and MMP9, thereby facilitating cell invasion in human trophoblast HTR-8/SVneo cells. Inhibition of ERK1/2 alleviated the expression of MMPs and trophoblast cell invasion after EG-VEGF treatment. In addition, primary cilia were observed in all the trophoblast cell lines tested and, more importantly, in human first-trimester placental tissue. The receptor of EG-VEGF, PROKR1, was detected in primary cilia. Depletion of IFT88, the intraflagellar transporter required for ciliogenesis, inhibited primary cilium growth, thereby ameliorating ERK1/2 activation, MMP upregulation, and trophoblast cell invasion promoted by EG-VEGF. These findings demonstrate a novel function of primary cilia in controlling EG-VEGF-regulated trophoblast invasion and reveal the underlying molecular mechanism. J. Cell. Physiol. 232: 1467–1477, 2017. © 2016 Wiley Periodicals, Inc.Primary cilium is a cellular antenna that conveys extracellular signals into a cellular response. EG-VEGF binds to PROKR1 locating on the cilium, and activates ERK1/2 signaling followed by upregulating MMP2 and MMP9 for facilitating cell invasion. We demonstrated that EG-VEGF regulated trophoblast cell invasion via primary cilium in the present study.
      PubDate: 2016-12-29T05:40:59.355062-05:
      DOI: 10.1002/jcp.25649
       
  • Increased Chondrogenic Potential of Mesenchymal Cells From Adipose Tissue
           Versus Bone Marrow-Derived Cells in Osteoarthritic In Vitro Models
    • Authors: Stefania Pagani; Veronica Borsari, Francesca Veronesi, Andrea Ferrari, Simona Cepollaro, Paola Torricelli, Giuseppe Filardo, Milena Fini
      Pages: 1478 - 1488
      Abstract: Primarily, to compare the behavior of human mesenchymal stem cells (MSCs) derived from bone marrow (hBMSCs) and adipose tissue (hADSCs) in an osteoarthritic (OA) microenvironment; secondly, to investigate the reaction of these cell types in two alternative in vitro culture systems, obtained by using TNFα and/or IL1β as inflammation mediators, or by using synovial fluid harvested by OA patients (OSF) to simulate the complex inflamed knee microenvironment. 3D micromass cultures of hBMSCs or hADSCs were grown in chondrogenic medium (CTR), in the presence of TNFα and/or IL1β, or synovial fluid from OA patients. After 1 month of culture, the chondrogenic differentiation of micromasses was evaluated by gene expression, matrix composition, and organization. Both hMSCs types formed mature micromasses in CTR, but a better response of hADSCs to the inflammatory environment was documented by micromass area and Bern score evaluations. The addition of OSF elicited a milder reaction than with TNFα and/or IL1β by both cell types, probably due to the presence of both catabolic and protective factors. In particular, SOX9 and ACAN gene expression and GAG synthesis were more abundant in hADSCs than hBMSCs when cultured in OSF. The expression of MMP1 was increased for both hMSCs in inflammatory conditions, but in particular by hBMSCs. hADSCs showed an increased chondrogenic potential in inflammatory culture systems, suggesting a better response of hADSCs in the OA environment, thus underlining the importance of appropriate in vitro models to study MSCs and potential advantages of using these cells for future clinical applications. J. Cell. Physiol. 232: 1478–1488, 2017. © 2016 Wiley Periodicals, Inc.Primary objective of this paper was to compare the behavior of human mesenchymal stem cells (MSCs) derived from bone marrow (hBMSCs) and adipose tissue (hADSCs) in an osteoarthritic (OA) microenvironment; secondly, to investigate the reaction of these cell types in two alternative in vitro culture systems, obtained by using TNFα and/or IL1β as inflammation mediators, or by using synovial fluid harvested by OA patients (OSF) to simulate the complex inflamed knee microenvironment.
      PubDate: 2016-10-26T09:35:34.193948-05:
      DOI: 10.1002/jcp.25651
       
  • Src Family Kinase Links Insulin Signaling to Short Term Regulation of
           Na,K-ATPase in Nonpigmented Ciliary Epithelium
    • Authors: Mohammad Shahidullah; Amritlal Mandal, Nicholas A. Delamere
      Pages: 1489 - 1500
      Abstract: Insulin has been shown to elicit changes of Na,K-ATPase activity in various tissues. Na,K-ATPase in the nonpigmented ciliary epithelium (NPE) plays a role in aqueous humor secretion and changes of Na,K-ATPase activity impact the driving force. Because we detect a change of NPE Na,K-ATPase activity in response to insulin, studies were carried out to examine the response mechanism. Ouabain-sensitive rubidium (Rb) uptake by cultured NPE cells, measured as a functional index of Na,K-ATPase-mediated inward potassium transport, was found to increase in cells exposed for 5 min to insulin. The maximally effective concentration was 100 nM. An intrinsic increase of Na,K-ATPase activity evident as a>2-fold increase in the rate of ouabain-sensitive ATP hydrolysis in homogenates obtained from cells exposed to 100 nM insulin for 5 min was also observed. Insulin-treated cells exhibited Akt, Src family kinase (SFK), ERK1/2, and p38 activation, all of which were prevented by a pI3 kinase inhibitor LY294002. The Rb uptake and Na,K-ATPase activity response to insulin both were abolished by PP2, an SFK inhibitor which also prevented p38 and ERK1/2 but not Akt activation. The Akt inhibitor MK-2206 did not change the Na,K-ATPase response to insulin. The findings suggest insulin activates pI3K-dependent Akt and SFK signaling pathways that are separate. ERK1/2 and p38 activation is secondary to and dependent on SFK activation. The increase of Na,K-ATPase activity is dependent on activation of the SFK pathway. The findings are consistent with previous studies that indicate a link between Na,K-ATPase activity and SFK signaling. J. Cell. Physiol. 232: 1489–1500, 2017. © 2016 Wiley Periodicals, Inc.Studies were conducted to examine the insulin response in ocular nonpigmented ciliary epithelium. Insulin elicited a pronounced short term increase in Na,K-ATPase activity and a complex signaling response that involved Akt (protein kinase B), Src family kinase (SFK), ERK1/2, and p38 MAPK all of which were dependent on PI3K activation. ERK1/2 and p38 MAPK activation were secondary to and dependent on SFK activation. The insulin-dependent increase in Na,K-ATPase activity was linked to SFK activation. In contrast, Akt activation has no discernable effect on Na,K-ATPase activity.
      PubDate: 2016-11-10T06:51:00.404527-05:
      DOI: 10.1002/jcp.25654
       
  • Neuroprotection and Blood–Brain Barrier Restoration by Salubrinal
           After a Cortical Stab Injury
    • Authors: M. Asunción Barreda-Manso; Natalia Yanguas-Casás, Manuel Nieto-Sampedro, Lorenzo Romero-Ramírez
      Pages: 1501 - 1510
      Abstract: Following a central nervous system (CNS) injury, restoration of the blood-brain barrier (BBB) integrity is essential for recovering homeostasis. When this process is delayed or impeded, blood substances and cells enter the CNS parenchyma, initiating an additional inflammatory process that extends the initial injury and causes so-called secondary neuronal loss. Astrocytes and profibrotic mesenchymal cells react to the injury and migrate to the lesion site, creating a new glia limitans that restores the BBB. This process is beneficial for the resolution of the inflammation, neuronal survival, and the initiation of the healing process. Salubrinal is a small molecule with neuroprotective properties in different animal models of stroke and trauma to the CNS. Here, we show that salubrinal increased neuronal survival in the neighbourhood of a cerebral cortex stab injury. Moreover, salubrinal reduced cortical blood leakage into the parenchyma of injured animals compared with injured controls. Adjacent to the site of injury, salubrinal induced immunoreactivity for platelet-derived growth factor subunit B (PDGF-B), a specific mitogenic factor for mesenchymal cells. This effect might be responsible for the increased immunoreactivity for fibronectin and the decreased activation of microglia and macrophages in injured mice treated with salubrinal, compared with injured controls. The immunoreactivity for PDGF-B colocalized with neuronal nuclei (NeuN), suggesting that cortical neurons in the proximity of the injury were the main source of PDGF-B. Our results suggest that after an injury, neurons play an important role in both, the healing process and the restoration of the BBB integrity. J. Cell. Physiol. 232: 1501–1510, 2017. © 2016 Wiley Periodicals, Inc.Salubrinal is neuroprotective in an animal model of cortical stab injury and facilitated the restoration of the BBB integrity, by inducing the neuronal production of PDGF-B. Our results suggest that neurons in the proximity of the lesion have a previously underrated, major role in the restoration of the BBB.
      PubDate: 2016-11-10T06:50:54.521177-05:
      DOI: 10.1002/jcp.25655
       
  • ADP-Induced Ca2+ Signaling and Proliferation of Rat Ventricular
           Myofibroblasts Depend on Phospholipase C-Linked TRP Channels Activation
           Within Lipid Rafts
    • Authors: Mariana Certal; Adriana Vinhas, Aurora Barros-Barbosa, Fátima Ferreirinha, Maria Adelina Costa, Paulo Correia-de-Sá
      Pages: 1511 - 1526
      Abstract: Nucleotides released during heart injury affect myocardium electrophysiology and remodeling through P2 purinoceptors activation in cardiac myofibroblasts. ATP and UTP endorse [Ca2+]i accumulation and growth of DDR-2/α-SMA-expressing myofibroblasts from adult rat ventricles via P2Y4 and P2Y2 receptors activation, respectively. Ventricular myofibroblasts also express ADP-sensitive P2Y1, P2Y12, and P2Y13 receptors as demonstrated by immunofluorescence confocal microscopy and western blot analysis, but little information exists on ADP effects in these cells. ADP (0.003–3 mM) and its stable analogue, ADPßS (100 μM), caused fast [Ca2+]i transients originated from thapsigargin-sensitive internal stores, which partially declined to a plateau sustained by capacitative Ca2+ entry through transient receptor potential (TRP) channels inhibited by 2-APB (50 μM) and flufenamic acid (100 μM). Hydrophobic interactions between Gq/11-coupled P2Y purinoceptors and TRP channels were suggested by prevention of the ADP-induced [Ca2+]i plateau following PIP2 depletion with LiCl (10 mM) and cholesterol removal from lipid rafts with methyl-ß-cyclodextrin (2 mM). ADP [Ca2+]i transients were insensitive to P2Y1, P2Y12, and P2Y13 receptor antagonists, MRS2179 (10μM), AR-C66096 (0.1 μM), and MRS2211 (10μM), respectively, but were attenuated by suramin and reactive blue-2 (100 μM) which also blocked P2Y4 receptors activation by UTP. Cardiac myofibroblasts growth and type I collagen production were favored upon activation of MRS2179-sensitive P2Y1 receptors with ADP or ADPßS (30 μM). In conclusion, ADP exerts a dual role on ventricular myofibroblasts: [Ca2+]i transients are mediated by fast-desensitizing P2Y4 receptors, whereas the pro-fibrotic effect of ADP involves the P2Y1 receptor activation. Data also show that ADP-induced capacitative Ca2+ influx depends on phospholipase C-linked TRP channels opening in lipid raft microdomains. J. Cell. Physiol. 232: 1511–1526, 2017. © 2016 Wiley Periodicals, Inc.The P2Y4R mainly mediates ADP-induced [Ca2+]i rise in rat ventricular myofibroblasts; ADP-induced fast [Ca2+]i rises depend on Ca2+ recruitment from internal stores; the late [Ca2+]i plateau depends on capacitative Ca2+ entry via TRP channels; PLC-activating P2Y4 receptors and TRP channels are assembled in lipid rafts; ADP promotes cell growth and collagen production predominantly via P2Y1 activation.
      PubDate: 2016-11-10T06:51:12.972294-05:
      DOI: 10.1002/jcp.25656
       
  • Functional Roles of Eph A-Ephrin A1 System in Endometrial Luminal
           Epithelial Cells During Early Pregnancy
    • Authors: Whasun Lim; Hyocheol Bae, Fuller W. Bazer, Gwonhwa Song
      Pages: 1527 - 1538
      Abstract: Eph and ephrin regulate diverse biological events such as proliferation, adhesion, migration, and angiogenesis through cell-to-cell interactions. However, little is known of their functional role and mechanisms of action in uterine endometrial cells. In the present study, we demonstrated the effects of the Eph and ephrin on interactions between blastocysts and endometrial luminal epithelial (pLE) cells in the pig that is regarded as an excellent biomedical animal model for research on the peri-implantation period of pregnancy. Results of this study indicated that among eight members of the Eph A family, expression of Eph A1, A2, A4, and A7 was strongly detected in endometrial epithelial cells during early pregnancy. Of these, for identification of signal transduction pathways induced by ephrin A1, a major ligand for Eph A, cell proliferation assays, and immunofluorescence and cell cycle regulation were analyzed following treatment of pLE cells with ephrin A1. Ephrin A1 stimulated proliferation of pLE cells as evidenced by abundant PCNA expression and an increase in the G2/M phase. Western blot analysis showed that ephrin A1 activated PI3K and MAPK signaling proteins in a time-dependent manner. Moreover, phosphorylation of AKT, ERK1/2, P38, and JNK proteins were suppressed by their inhibitors wortmannin, U0126, SB203580, and SP600125, respectively. Also, phosphor-AKT was reduced by ERK1/2 and P38 inhibitors. Ephrin A1-induced proliferation and migration of pLE cells was also blocked by those inhibitors. Collectively, these results suggest that ephrin A1 enhances interactions between porcine blastocysts and endometrial luminal epithelial cells by activating PI3K and MAPK signal transduction pathways. J. Cell. Physiol. 232: 1527–1538, 2017. © 2016 Wiley Periodicals, Inc.Eph A-ephrin A1 system plays an important role in uterine receptivity and improvement of litter size through PI3K/AKT and MAPK signal transduction pathways during the peri-implantation period of pregnancy.
      PubDate: 2016-11-10T06:45:30.404277-05:
      DOI: 10.1002/jcp.25659
       
  • E-Cigarette Vapor Induces an Apoptotic Response in Human Gingival
           Epithelial Cells Through the Caspase-3 Pathway
    • Authors: Mahmoud Rouabhia; Hyun Jin Park, Abdelhabib Semlali, Andrew Zakrzewski, Witold Chmielewski, Jamila Chakir
      Pages: 1539 - 1547
      Abstract: Electronic cigarettes represent an increasingly significant proportion of today's consumable tobacco products. E-cigarettes contain several chemicals which may promote oral diseases. The aim of this study was to investigate the effect of e-cigarette vapor on human gingival epithelial cells. Results show that e-cigarette vapor altered the morphology of cells from small cuboidal form to large undefined shapes. Both single and multiple exposures to e-cigarette vapor led to a bulky morphology with large faint nuclei and an enlarged cytoplasm. E-cigarette vapor also increased L-lactate dehydrogenase (LDH) activity in the targeted cells. This activity was greater with repeated exposures. Furthermore, e-cigarette vapor increased apoptotic/necrotic epithelial cell percentages compared to that observed in the control. Epithelial cell apoptosis was confirmed by TUNEL assay showing that exposure to e-cigarette vapor increased apoptotic cell numbers, particularly after two and three exposures. This negative effect involved the caspase-3 pathway, the activity of which was greater with repeated exposure and which decreased following the use of caspase-3 inhibitor. The adverse effects of e-cigarette vapor on gingival epithelial cells may lead to dysregulated gingival cell function and result in oral disease. J. Cell. Physiol. 232: 1539–1547, 2017. © 2016 Wiley Periodicals, Inc.E-cigarette vapor mediated gingival epithelial cell activity by promoting apoptosis which took place through an apoptosis pathway involving caspase-3. The effects of e-cigarette vapor on gingival epithelial cells may compromise epithelial tissue, resulting in periodontal disease development and potentiating inflammation resulting from the tissue damage.
      PubDate: 2016-11-30T10:35:54.111849-05:
      DOI: 10.1002/jcp.25677
       
  • Notch Signaling Pathway Regulates Angiogenesis via Endothelial Cell in 3D
           Co-Culture Model
    • Authors: Dan Zhao; Changyue Xue, Shiyu Lin, Sirong Shi, Qianshun Li, Mengting Liu, Xiaoxiao Cai, Yunfeng Lin
      Pages: 1548 - 1558
      Abstract: This study aimed to investigate the role of Notch signaling pathway for angiogenesis in a three-dimensional (3D) collagen gel model with co-culture of adipose-derived stromal cells (ASCs) and endothelial cells (ECs). A 3D collagen gel model was established in vitro by implanting both ASCs from green fluorescent protein-labeled mouse and ECs from red fluorescent protein-labeled mouse, and the phenomena of angiogenesis with Notch signaling inducer Jagged1, inhibitor DAPT and PBS, respectively were observed by confocal laser scanning microscopy. Semi-quantitative PCR and immunofluorescent staining were conducted to detect expressions of angiogenesis-related genes and proteins. Angiogenesis in the co-culture gels was promoted by Jagged1 treatment while attenuated by DAPT treatment, compared to control group. In co-culture system of ASCs and ECs, the gene expressions of VEGFA, VEGFB, Notch1, Notch2, Hes1, Hey1, VEGFR1,and the protein expression of VEGFA, VEGFB, Notch1, Hes1, Hey1 were increased by Jagged1 treatment and decreased by DAPT treatment in ECs. And the result of VEGFR3 was the opposite. However, the same results did not appear completely in ASCs. These results revealed the VEGFA/B-Notch1/2-Hes1/Hey1- VEGFR1/3 signal axis played an important role in angiogenesis when ASCs and ECs were co-cultured in a 3D collagen gel model. J. Cell. Physiol. 232: 1548–1558, 2017. © 2016 Wiley Periodicals, Inc.VEGFA/B-Notch1/2-Hes1/Hey1- VEGFR1/3 signal axis played an important role in angiogenesis when ASCs and ECs were co-cultured in a 3D collagen gel model.
      PubDate: 2016-11-20T15:45:44.326649-05:
      DOI: 10.1002/jcp.25681
       
  • S100B + A1 CELISA: A Novel Potency Assay and Screening Tool for
           Redifferentiation Stimuli of Human Articular Chondrocytes
    • Authors: Jose Diaz-Romero; Sibylle Kürsener, Sandro Kohl, Dobrila Nesic
      Pages: 1559 - 1570
      Abstract: During monolayer expansion, a necessary step in autologous chondrocyte implantation, human articular chondrocytes (HAC) dedifferentiate and lose their capacity to produce stable hyaline cartilage. Determining HAC potency and learning how to trigger their redifferentiation would improve cell-based cartilage regeneration therapies. We previously identified S100B and S100A1 proteins as markers of HAC redifferentiation potential. Here, we aimed to: (i) demonstrate a correlation between S100B + A1-positive HAC in monolayer culture and their neochondrogenesis capacity in pellet culture; (ii) develop an S100B + A1 cell-based ELISA, and (iii) prove that S100B + A1 induction in HAC increases their chondrogenic capacity. Expression patterns of S100A1 and S100B were investigated in HAC during dedifferentiation (monolayer) or redifferentiation (pellet or high-osmolarity/BMP4 treatment in monolayer) using qRT-PCR, immunocytochemistry, or immunohistochemistry. A cell-based ELISA (CELISA) was developed as a 96-well microplate multiplex assay to measure S100B + A1 (chondrogenesis), alkaline phosphatase (hypertrophy), and DNA amount (normalization), and applied to HAC, bone marrow-derived mesenchymal stem cells and the chondrocytic cell line ATDC5. The direct correlation between the percentage of S100B + A1-positive HAC in monolayer and their neochondrogenesis in pellets validates S100B + A1 as a marker of chondrogenic potency. The S100B + A1-CELISA accurately determines HAC differentiation status, allows identification of chondrogenic stimuli, and permits the simultaneous monitoring of the undesirable hypertrophic phenotype. This novel assay offers a high-throughput, comprehensive and versatile approach for measuring cell chondrogenic potency and for identifying redifferentiation factors/conditions. HAC improved neochondrogenesis in pellets—induced with high-osmolarity and BMP4 treatment in monolayer—suggests that cell instruction prior to implantation may improve cartilage repair. J. Cell. Physiol. 232: 1559–1570, 2017. © 2016 Wiley Periodicals, Inc.The novel S100B + A1-CELISA offers a high-throughput, comprehensive and versatile approach for measuring cell chondrogenic potency and identifying redifferentiation factors/conditions to improve cartilage repair. Using a combination of high-osmolarity medium and BMP4 treatment uncovered with this assay, we demonstrate that the induction of chondrocyte redifferentiation in monolayer improves their chondrogenic potential in pellets.
      PubDate: 2016-11-30T10:35:40.68692-05:0
      DOI: 10.1002/jcp.25682
       
  • A Real-World Multicentre Retrospective Study of Paclitaxel-Bevacizumab and
           Maintenance Therapy as First-Line for HER2-Negative Metastatic Breast
           Cancer
    • Authors: Teresa Gamucci; Lucia Mentuccia, Clara Natoli, Isabella Sperduti, Alessandra Cassano, Andrea Michelotti, Luigi Di Lauro, Domenico Sergi, Alessandra Fabi, Maria G. Sarobba, Paolo Marchetti, Maddalena Barba, Emanuela Magnolfi, Marcello Maugeri-Saccà, Ernesto Rossi, Valentina Sini, Antonino Grassadonia, Domenica Pellegrini, Antonino Astone, Cecilia Nisticò, Franco Angelini, Angela Vaccaro, Arianna Pellegrino, Claudia De Angelis, Michela Palleschi, Luca Moscetti, Ilaria Bertolini, Simonetta Buglioni, Antonio Giordano, Laura Pizzuti, Patrizia Vici
      Pages: 1571 - 1578
      Abstract: Bevacizumab in combination with taxanes in HER2-negative metastatic breast cancer (MBC) patients has shown improved progression-free survival (PFS), despite the lack of clear overall survival (OS) benefit. We performed a retrospective analysis to evaluate the impact of paclitaxel-bevacizumab and of maintenance therapy with bevacizumab (BM) and endocrine therapy (ET) in the real-world practice. We identified 314 HER2-negative MBC patients treated in 12 cancer centers. Overall, the median PFS and OS were 14 and 40 months, respectively. Among the 254 patients potentially eligible for BM, 183 received BM after paclitaxel discontinuation until progression/toxicity. PFS and OS were improved in patients who had received BM in comparison with those potentially eligible but who did not receive BM (P
      PubDate: 2016-11-30T10:40:55.407035-05:
      DOI: 10.1002/jcp.25685
       
 
 
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