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Journal Cover   Journal of Cellular Physiology
  [SJR: 1.948]   [H-I: 125]   [3 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  [1610 journals]
  • Pathophysiology of Corneal Dystrophies: From Cellular Genetic Alteration
           to Clinical Findings
    • Authors: Marta Sacchetti; Ilaria Macchi, Alessandro Tiezzi, Maurizio La Cava, Giacomina Massaro‐Giordano, Alessandro Lambiase
      Abstract: Corneal dystrophies are a heterogeneous group of bilateral, inherited, rare diseases characterized by slowly progressive corneal opacities, that lead to visual impairment. Most of them have an autosomal dominant pattern of inheritance with variable expressivity, but new mutations have been described. Many corneal dystrophies have been genetically characterized and the specific gene mutations identified, such as for the epithelial–stromal TGFBI dystrophies. Current classification systems identified four main groups of corneal dystrophies based on clinical, histologic and genetic information. Diagnosis is performed during a routine ophthalmic examination that shows typical cellular abnormalities of the corneal epithelium, stroma or endothelium. Disease progression should be carefully monitored to decide the proper clinical management. The treatment of corneal dystrophies is variable, depending on symptoms, clinical course, severity and type of dystrophy. Management aimed to reduce symptoms and to improve vision, includes different surgical approaches. Novel cellular and genetic therapeutic approaches are under evaluation. This article is protected by copyright. All rights reserved
      PubDate: 2015-06-23T18:46:57.198342-05:
      DOI: 10.1002/jcp.25082
       
  • Highlights: Volume 230, Number 10
    • PubDate: 2015-06-23T17:18:50.110703-05:
      DOI: 10.1002/jcp.24767
       
  • Editor's Choice
    • PubDate: 2015-06-23T17:18:45.143714-05:
      DOI: 10.1002/jcp.25063
       
  • Journal of Cellular Physiology: Volume 230, Number 10, October 2015
    • Abstract: Cover: Modeling of compound BCI‐121 in complex with SMYD3 histone methyltransferase. See article by Peserico et al. on pages 2447–2460.
      PubDate: 2015-06-23T17:18:38.571829-05:
      DOI: 10.1002/jcp.24765
       
  • Table of Contents: Volume 230, Number 10
    • PubDate: 2015-06-23T17:18:35.921979-05:
      DOI: 10.1002/jcp.24766
       
  • Is the Reg3α (HIP/PAP) Protein Really an Obesogenic Factor?
    • Authors: Patrick Gonzalez; Nicolas Moniaux, Christian Bréchot, Jamila Faivre
      Abstract: We would like to point out an internal inconsistency in the data presented by V. Secq et al. in the article entitled “PAP/HIP protein is an obesogenic factor” published in the February 2014 issue of “Journal of Cellular Physiology”, which cast serious doubt on the validity of the statement serving as the title of the article. We, and others, including J. Iovanna and his collaborators, have long shown that the C‐type lectin PAP/HIP (also known as HIP/PAP or Reg3α) has anti‐inflammatory and antioxidant activities in a number of cell types, among which pancreatic cells, hepatocytes and neurons (Gironella et al., 2005; Closa et al., 2007; Moniaux et al., 2011; Kapur et al., 2012; Haldipur et al., 2014) and that, therefore, it might help cure, or prevent, inflammatory and metabolic diseases. Given its envisaged clinical uses, the question of whether PAP/HIP significantly modifies metabolism toward weight increase is essential and must be subjected to rigorous scrutiny. This article is protected by copyright. All rights reserved
      PubDate: 2015-06-19T17:57:14.333248-05:
      DOI: 10.1002/jcp.25046
       
  • Critical Role of AMPK/FoxO3A Axis in Globular Adiponectin‐Induced
           Cell Cycle Arrest and Apoptosis in Cancer Cells
    • Authors: Anup Shrestha; Saroj Nepal, Mi Jin Kim, Jae Hoon Chang, Sang‐Hyun Kim, Gil‐Saeng Jeong, Chul‐Ho Jeong, Gyu Hwan Park, Sunghee Jung, Jaecheong Lim, Eunha Cho, Soyoung Lee, Pil‐Hoon Park
      Abstract: Adiponectin predominantly secreted from adipose tissue has exhibited potent anti‐proliferative properties in cancer cells via modulating cell cycle and apoptosis. FoxO3A, a Forkhead box O member of the transcription factor, plays a critical role in modulating expression of genes involved in cell death and/or survival. In this study, we investigated the role of FoxO3A signaling in anti‐cancer activities of adiponectin. Herein, we have shown that treatment with globular adiponectin (gAcrp) increases p27 but decreases cyclinD1 expression in human hepatoma (HepG2) and breast (MCF‐7) cancer cells. Gene ablation of FoxO3A prevented gAcrp‐induced increase in p27 and decreased in cyclin D1 expression, and further ameliorated cell cycle arrest by gAcrp, indicating a critical role of FoxO3A in gAcrp‐induced cell cycle arrest of cancer cells. Moreover, treatment with gAcrp also induced caspase‐3/7 activation and increased Fas ligand (FasL) expression in both HepG2 and MCF‐7 cells. Transfection with FoxO3A siRNA inhibited gAcrp‐induced caspase‐3/7 activation and FasL expression, suggesting that FoxO3A signaling also plays an important role in gAcrp‐induced apoptosis of cancer cells. We also found that gene silencing of AMPK prevented gAcrp‐induced nuclear translocation of FoxO3A in HepG2 and MCF‐7 cells. In addition, suppression of AMPK also blocked gAcrp‐induced cell cycle arrest and further attenuated gAcrp‐induced caspase‐3/7 activation, indicating that AMPK signaling plays a pivotal role in both gAcrp‐induced cell cycle arrest and apoptosis via acting as an upstream signaling of FoxO3A. Taken together, our findings demonstrated that AMPK/FoxO3A axis plays a cardinal role in anti‐proliferative effect of adiponectin in cancer cells. This article is protected by copyright. All rights reserved
      PubDate: 2015-06-18T18:54:36.875656-05:
      DOI: 10.1002/jcp.25080
       
  • Platelet‐Derived CCL5 Regulates CXC Chemokine Formation and
           Neutrophil Recruitment in Acute Experimental Colitis
    • Authors: Changhui Yu; Songen Zhang, Yongzhi Wang, Su Zhang, Lingtao Luo, Henrik Thorlacius
      Abstract: Accumulating data suggest that platelets not only regulate thrombosis and haemostasis but also inflammatory processes. Platelets contain numerous potent pro‐inflammatory compounds, including the chemokines CCL5 and CXCL4 although their role in acute colitis remains elusive. The aim of this study was to examine the role of platelets and platelet‐derived chemokines in acute colitis. Acute colitis was induced in female Balb/c mice by administration of 5% dextran sodium sulphate (DSS) for five days. Animals received a platelet‐depleting, anti‐CCL5, anti‐CXCL4 or a control antibody prior to DSS challenge. Colonic tissue was collected for quantification of myeloperoxidase (MPO) activity, CXCL5, CXCL2, interleukin‐6 (IL‐6) and CCL5 levels as well as morphological analyses. Platelet depletion reduced tissue damage and clinical disease activity index in DSS‐exposed animals. Platelet depletion not only reduced levels of CXCL2 and CXCL5 but also levels of CCL5 in the inflamed colon. Immunoneutralization of CCL5 but not CXCL4 reduced tissue damage, CXC chemokine expression and neutrophil recruitment in DSS‐treated animals. These findings show that platelets play a key role in acute colitis by regulating CXC chemokine generation, neutrophil infiltration and tissue damage in the colon. Moreover, our results suggest that platelet‐derived CCL5 is an important link between platelet activation and neutrophil recruitment in acute colitis. This article is protected by copyright. All rights reserved
      PubDate: 2015-06-18T18:54:10.466808-05:
      DOI: 10.1002/jcp.25081
       
  • Genetic and Functional Analysis of Polymorphisms in the Human Dopamine
           Receptor and Transporter Genes in Small Cell Lung Cancer
    • Authors: Emanuela Cherubini; Arianna Di Napoli, Alessia Noto, Giorgia Amira Osman, Maria Cristina Esposito, Salvatore Mariotta, Rossella Sellitri, Luigi Ruco, Giuseppe Cardillo, Gennaro Ciliberto, Rita Mancini, Alberto Ricci
      Abstract: The regulatory role of dopamine (DA) in endocrine, cardiovascular and renal functions has been extensively studied and used for clinical purposes. More recently DA has been indicated as a regulatory molecule for immune cells and malignant cell proliferation. We assessed the expression and the functional role DA, DA receptors and transporters in primary small cell lung cancer (SCLC). By HPLC DA plasma levels were more elevated in SCLC patients in comparison with NSCLC patients and healthy controls. SCLC cell expressed DA D1‐ and D2‐like receptors and membrane and vesicular transporters at protein and mRNA levels. We also investigated the effects of independent D1‐ or D2‐like receptor stimulation on SCLC cell cultures. DA D1 receptor agonist SKF38393 induced the increase of cAMP levels and DARPP‐32 protein expression without affecting SCLC growth rate. Cell treatment with the DA D1 receptor antagonist SCH23390 inhibited SKF38393 effects. In contrast, the DA D2 receptor agonist quinpirole (10μM) counteracted, in a dose and time dependent way, SCLC cell proliferation, it did not affect cAMP levels and decreased phosphorylated AKT that was induced by DA D2 receptor antagonist sulpiride. However, in only one SCLC line, stimulation of DA D2 receptor failed to inhibit cell proliferation in vitro. This effect was associated to the existence of rs6275 and rs6277 polymorphisms in the D2 gene. These results gave more insight into DA control of lung cancer cell behavior and suggested the existence of different SCLC phenotypes. This article is protected by copyright. All rights reserved
      PubDate: 2015-06-16T17:46:14.085232-05:
      DOI: 10.1002/jcp.25079
       
  • Aquaporin‐1 Plays Important Role in Proliferation by Affecting Cell
           Cycle Progression
    • Authors: Ana Galán‐Cobo; Reposo Ramírez‐Lorca, Juan José Toledo‐Aral, Miriam Echevarría
      Abstract: Aquaporin‐1 (AQP1) has been associated with tumor development. Here, we investigated how AQP1 may affect cell proliferation. The proliferative rate of adult carotid body (CB) cells, known to proliferate under chronic hypoxia, was analyzed in wild‐type (AQP1 +/ +) and knock out (AQP1 ‐/‐) mice, maintained in normoxia or exposed to hypoxia while BrdU was administered. Fewer numbers of total BrdU+ and TH‐BrdU+ cells were observed in AQP1 ‐/‐ mice, indicating a role for AQP1 in CB proliferation. Then, by flow cytometry, cell cycle state and proliferation of cells overexpressing AQP1 were compared to those of wild‐type cells. In the AQP1‐overexpressing cells, we observed higher cell proliferation and percentages of cells in phases S and G2/M and fewer apoptotic cells after nocodazole treatment were detected by annexin V staining. Also in these cells, proteomic assays showed higher expression of cyclin D1 and E1 and microarray analysis revealed changes in many cell proliferation‐related molecules, including, Zeb 2, Jun, NF‐kβ, Cxcl9, Cxcl10, TNF, and the TNF receptor. Overall, our results indicate that the presence of AQP1 modifies the expression of key cell cycle proteins apparently related to increases in cell proliferation. This contributes to explaining the presence of AQP1 in many different tumors. This article is protected by copyright. All rights reserved
      PubDate: 2015-06-16T17:43:07.293339-05:
      DOI: 10.1002/jcp.25078
       
  • Dexamethasone Downregulates SLC7A5 Expression and Promotes Cell Cycle
           Arrest, Autophagy and Apoptosis in BeWo Cells
    • Authors: Bin He; Nana Zhang, Ruqian Zhao
      Abstract: Synthetic glucocorticoids (GCs) such as dexamethasone (Dex) are widely given to pregnant women to induce maturation and improve viability of preterm infants. Despite the beneficial effects, synthetic GCs have adverse effects on placental growth and nutrient transport system. However, the molecular mechanisms involved in these events remain unknown. Here we use a human placental choriocarcinoma cell line (BeWo) as model to explore the pathway linking amino acids transport with cell viability under Dex challenge. BeWo cells treated with Dex (100 nM) for 24 h demonstrated G1/S cell cycle arrest together with enhanced autophagy and apoptosis. Concurrently, the amino acid carrier SLC7A5 was down‐regulated in association with impaired cellular amino acids uptake and inhibition of mammalian target of rapamycin (mTOR) signaling. Similar cellular responses were observed in BeWo cells treated with BCH, a classical System L inhibitor which inactivates SLC7A5. The glucocorticoid receptor (GR) antagonist RU486 was able to diminish Dex‐induced translocation of GR into nucleus and to abolish these effects. Furthermore, Dex treatment significantly promoted the binding of GR to the proximal promoter sequence of SLC7A5 gene. Taken together, our results show that Dex downregulates SLC7A5 expression via GR‐mediated transrepression. The impaired amino acids uptake leads to inhibition of mTOR signaling which in turn causes inhibited proliferation and enhanced autophagy and apoptosis in BeWo cells. These findings indicate that SLC7A5 mediates the effect of Dex on cell viability, thus providing a novel molecular target for the prevention and treatment of Dex‐induced cell cycle arrest and apoptosis in placental cells. This article is protected by copyright. All rights reserved
      PubDate: 2015-06-10T18:47:13.684676-05:
      DOI: 10.1002/jcp.25076
       
  • G‐CSF Contributes at the Healing of Tunica Media of
           Arteriotomy‐Injured Rat Carotids by Promoting Differentiation of
           Vascular Smooth Muscle Cells
    • Authors: Barbara Rinaldi; Mauro Finicelli, Maria Donniacuo, Giovanni Di Bernardo, Giulia Gritti, Stefania Del Gaudio, Amalia Forte, Gianfranco Peluso, Marilena Cipollaro, Francesco Rossi, Umberto Galderisi
      Abstract: Restenosis is a complex pathophysiological disease whose causative mechanisms are not fully understood. Previous studies allowed us to demonstrate the efficacy of bone marrow mesenchymal stromal cells (MSCs) transplantation in limiting the pathophysiological remodeling in a model of arteriotomy‐induced (re)stenosis. In the current research we studied the effectiveness of G‐CSF treatment on male rate rats that were subjected carotid arteriotomy in order to evaluate a potentially effective non‐invasive strategy that recapitulates the MSC‐mediated recovery of injured vessels. WKY male rats were subjected carotid arteriotomy and given a nine day treatment (3 days pre‐ to 6 days post‐arteriotomy) with G‐CSF or saline. Carotids were harvested 7 and 30 days following arteriotomy (early‐ and late‐phase, respectively). Although morphometrical analysis did not reveal differences in lumen narrowing between G‐CSF‐ and PBS‐carotids 30 days following arteriotomy, we detected a noticeable conservative effect of G‐CSF treatment on vascular wall morphology. Histological and molecular analysis revealed an increase in cellularity within the tunica media with a concomitant increase of the VSMCs differentiation markers both at early‐ and late‐phases of (re)stenotic response in G‐CSF‐treated carotids (Sm22‐alpha, Myocd and Smtn). These findings were accompanied by the downregulation of oxidative stress‐related genes in G‐CSF‐injured rats. The effect exerted by G‐CSF in our model of arteriotomy‐induced (re)stenosis seemed support the recovery of the architecture of the tunica media of injured vessels by: i) inducing VSMCs differentiation; and ii) limiting the oxidative‐stress response induced by arteriotomy. This article is protected by copyright. All rights reserved
      PubDate: 2015-06-10T18:04:03.48673-05:0
      DOI: 10.1002/jcp.25074
       
  • Pathways Implicated in Tadalafil Amelioration of Duchenne Muscular
           Dystrophy
    • Authors: Valeria De Arcangelis; Georgios Strimpakos, Francesca Gabanella, Nicoletta Corbi, Siro Luvisetto, Armando Magrelli, Annalisa Onori, Claudio Passananti, Cinzia Pisani, Sophie Rome, Cinzia Severini, Fabio Naro, Elisabetta Mattei, Maria Grazia Di Certo, Lucia Monaco
      Abstract: Numerous therapeutic approaches for Duchenne and Becker Muscular Dystrophy (DMD and BMD), the most common X‐linked muscle degenerative disease, have been proposed. So far, the only one showing a clear beneficial effect is the use of corticosteroids. Recent evidence indicates an improvement of dystrophic cardiac and skeletal muscles in the presence of sustained cGMP levels secondary to a blocking of their degradation by phosphodiesterase 5 (PDE5). Due to these data, we performed a study to investigate the effect of the specific PDE5 inhibitor, tadalafil, on dystrophic skeletal muscle function. Chronic pharmacological treatment with tadalafil has been carried out in mdx mice. Behavioral and physiological tests, as well as histological and biochemical analyses, confirmed the efficacy of the therapy. We then performed a microarray‐based genomic analysis to assess the pattern of gene expression in muscle samples obtained from the different cohorts of animals treated with tadalafil. This scrutiny allowed us to identify several classes of modulated genes. Our results show that PDE5 inhibition can ameliorate dystrophy by acting at different levels. Tadalafil can lead to 1) increased lipid metabolism; 2) a switch towards slow oxidative fibers driven by the up‐regulation of PGC‐1α; 3) an increased protein synthesis efficiency; 4) a better actin network organization at Z‐disk. This article is protected by copyright. All rights reserved
      PubDate: 2015-06-10T18:03:22.713185-05:
      DOI: 10.1002/jcp.25075
       
  • Tamoxifen Inhibits TGF‐β‐Mediated Activation of
           Myofibroblasts by Blocking Non‐Smad Signaling through ERK1/2
    • Authors: Jon M. Carthy; Anders Sundqvist, Angelos Heldin, Hans van Dam, Dimitris Kletsas, Carl‐Henrik Heldin, Aristidis Moustakas
      Abstract: TGF‐β is a multifunctional cytokine which stimulates the differentiation of fibroblasts into myofibroblasts. Myofibroblasts are critical for normal wound healing, but also accumulate pathologically in a number of chronic inflammatory conditions where they are key contributors to aberrant tissue remodeling and fibrosis, and in cancer stroma. In the current study, we identified a role for tamoxifen as a potent inhibitor of the TGF‐β‐mediated activation of primary human skin and breast fibroblasts. Our data indicate that tamoxifen does not interfere with canonical Smad signaling downstream of TGF‐β but rather blocks non‐Smad signaling through ERK1/2 MAP‐kinase and the AP‐1 transcription factor FRA2. We further demonstrate by siRNA‐mediated knockdown that FRA2 is critical for the induced expression of myogenic proteins in response to TGF‐β. Functionally, TGF‐β‐stimulated fibroblast‐mediated contraction of collagen gels was impaired in the presence of tamoxifen. Altogether, these data demonstrate that tamoxifen prevents myofibroblast differentiation and therefore may provide therapeutic benefits to patients suffering from chronic inflammatory conditions or cancer. This article is protected by copyright. All rights reserved
      PubDate: 2015-06-10T18:02:58.161846-05:
      DOI: 10.1002/jcp.25049
       
  • Translating Regenerative Biomaterials into Clinical Practice
    • Authors: Edward T Stace; Stephanie G Dakin, Pierre‐Alexis Mouthuy, Prof Andrew J Carr
      Abstract: Globally health care spending is increasing unsustainably. This is especially true of the treatment of Musculoskeletal (MSK) disease where in the USA the MSK disease burden has doubled over the last 15 years. With an ageing and increasingly obese population, the surge in MSK related spending is only set to worsen. Despite increased funding, research and attention to this pressing health need, little progress has been made towards novel therapies. Tissue Engineering and Regenerative Medicine (TERM) strategies could provide the solutions required to mitigate this mounting burden. Biomaterial based treatments in particular present a promising field of potentially cost‐effective therapies. However, the translation of a scientific development to a successful treatment is fraught with difficulties. These barriers have so far limited translation of TERM science into clinical treatments. It is crucial for primary researchers to be aware of the barriers currently restricting the progression of science to treatments. Researchers need to act prospectively to ensure the clinical, financial and regulatory hurdles which seem so far removed from laboratory science do not stall or prevent the subsequent translation of their idea into a treatment. The aim of this review is to explore the development and translation of new treatments. Increasing the understanding of these complexities and barriers amongst primary researchers could enhance the efficiency of biomaterial translation. This article is protected by copyright. All rights reserved
      PubDate: 2015-06-09T02:41:27.983892-05:
      DOI: 10.1002/jcp.25071
       
  • Core Binding Factorβ Plays a Criticalrole During Chondrocyte
           Differentiation
    • Authors: Na‐Rae Park; Kyung‐Eun Lim, Min‐Su Han, Xiangguo Che, Clara YongjooPark, Jung‐Eun Kim, Ichiro Taniuchi, Suk‐Chul Bae, Je‐Yong Choi
      Abstract: Core binding factorβ (Cbfβ) is a partner protein of Runx family transcription factors with minimally characterized function in cartilage. Here we address the role of Cbfβ in cartilage by generating chondrocyte‐specific Cbfβ‐deficient mice (CbfbΔch/Δch) from Cbfb‐floxed mice crossed with mice expressing Cre from the Col2a1 promoter. Cbfb▵ch/▵ch mice diedsoon after birth and exhibiteddelayed endochondral bone formation,shorter appendicular skeleton length with increased proliferative chondrocytes,and nearly absent hypertrophic chondrocyte zones. Immunohistochemicaland quantitative real‐time PCR analyses showed that the proliferative chondrocytes increasedand the expression of markers of chondrocyte maturation at the growth plates, including Runx2, osterix, and osteopontin, were significantlydiminishedin Cbfb▵ch/▵ch mice compared to wild type mice. With regard to signaling pathways, both PTHrP‐Ihh and BMP signaling were compromised inCbfb▵ch/▵ch mice. Mechanistically, Cbfβdeficiency in chondrocytes caused a decrease of protein levels of Runxtranscription factors by accelerating polyubiquitination‐mediated proteosomal degradation in vitro. Indeed, Runx2 and Runx3, but notRunx1,decreased in Cbfb▵ch/▵ch mice. Collectively, these findings indicate that Cbfβplays a critical role for chondrocyte differentiationthrough stabilizingRunx2 and Runx3protein in cartilage This article is protected by copyright. All rights reserved
      PubDate: 2015-06-08T09:19:12.920884-05:
      DOI: 10.1002/jcp.25068
       
  • Characterization of Human Dermal Fibroblasts in Fabry Disease
    • Authors: J. Lakomá; V. Donadio, R. Liguori, M. Caprini
      Abstract: Fabry disease (FD) is a hereditary X‐linked metabolic lysosomal storage disorder due to insufficient amounts or a complete lack of the lysosomal enzyme α‐galactosidase A (α − GalA). The loss of α‐GalA activity leads to an abnormal accumulation of globotriaosylceramide (Gb3) in lysosomes and other cellular components of different tissues and cell types, affecting the cell function. However, whether these biochemical alterations also modify functional processes associated to the cell mitotic ability is still unknown. The goal of the present study was to characterize lineages of human dermal fibroblasts (HDFs) of FD patients and healthy controls focusing on Gb3 accumulation, expression of chloride channels that regulate proliferation, and proliferative activity. The biochemical and functional analyses indicate the existence of quantitative differences in some but not all the parameters of cytoskeletal organization, proliferation and differentiation processes. This article is protected by copyright. All rights reserved
      PubDate: 2015-06-08T09:18:30.179422-05:
      DOI: 10.1002/jcp.25072
       
  • Participation of TNF‐α in Inhibitory Effects of Adipocytes on
           Osteoblast Differentiation
    • Authors: Robrigo P. F. Abuna; Fabiola S. de Oliveira, Thiago De S. Santos, Thais R. Guerra, Adalberto L. Rosa, Marcio M. Beloti
      Abstract: Mesenchymal stem cells from bone marrow (BM‐MSCs) and adipose tissue (AT‐MSCs) are attractive tools for cell‐based therapies to repair bone tissue. In this study, we investigated the osteogenic and adipogenic potential of BM‐MSCs and AT‐MSCs as well as the effect of crosstalk between osteoblasts and adipocytes on cell phenotype expression. Rat BM‐MSCs and AT‐MSCs were cultured either in growth, osteogenic or adipogenic medium to evaluate osteoblast and adipocyte differentiation. Additionally, osteoblasts and adipocytes were indirectly co‐cultured to investigate the effect of adipocytes on osteoblast differentiation and vice versa. BM‐MSCs and AT‐MSCs exhibit osteogenic and adipogenic potential under non‐differentiation‐inducing conditions. When exposed to osteogenic medium, BM‐MSCs exhibited higher expression of bone markers compared with AT‐MSCs. Conversely, under adipogenic conditions, AT‐MSCs displayed higher expression of adipose tissue markers compared with BM‐MSCs. The presence of adipocytes as indirect co‐culture repressed the expression of the osteoblast phenotype, whereas osteoblasts did not exert remarkable effect on adipocytes. The inhibitory effect of adipocytes on osteoblasts was due to the release of tumor necrosis factor alpha (TNF‐α) in culture medium by adipocytes. Indeed, the addition of exogenous TNF‐α in culture medium repressed the differentiation of BM‐MSCs into osteoblasts mimicking the indirect co‐culture effect. In conclusion, our study showed that BM‐MSCs are more osteogenic while AT‐MSCs are more adipogenic. Additionally, we demonstrated the key role of TNF‐α secreted by adipocytes on the inhibition of osteoblast differentiation. Thus, we postulate that the higher osteogenic potential of BM‐MSCs makes them the first choice for inducing bone repair in cell‐based therapies. This article is protected by copyright. All rights reserved
      PubDate: 2015-06-08T09:17:38.647095-05:
      DOI: 10.1002/jcp.25073
       
  • ATP release and P2Y Receptor Signalling are Essential for Keratinocyte
           Galvanotaxis
    • Authors: Aimie Riding; Christine E. Pullar
      Abstract: Repair to damaged tissue requires directional cell migration to heal the wound. Immediately upon wounding an electrical guidance cue is created with the cathode of the electric field (EF) located at the center of the wound. Previous research has demonstrated directional migration of keratinocytes towards the cathode when an EF of physiological strength (100–150 mV/mm) is applied in vitro, but the “sensor” by which keratinocytes sense the EF remains elusive. Here we use a customised chamber design to facilitate the application ofa direct current (DC) EF of physiological strength (100 mV/mm) to keratinocytes whilst pharmacologically modulating the activation ofboth connexin hemichannels and purinergic receptorsto determine their role inEF‐mediated directional keratinocyte migration, galvanotaxis. In addition, keratinocytes were exposed to DiSCAC2(3) dye to visualize membrane potential changes within the cell upon exposure to the applied DC EF. Here we unveil ATP‐medicated mechanisms that underpin the initiation of keratinocyte galvanotaxis. The application of a DC EF of 100 mV/mm releases ATP via hemichannels activatinga subset of purinergic P2Y receptors, locally, to initiate the directional migration of keratinocytes towards the cathodein vitro, the center of the woundin vivo. The delineation of the mechanisms underpinning galvanotaxis extends our understanding of this endogenous cue and will facilitate the optimization and wider use of EF devices for chronic wound treatment. This article is protected by copyright. All rights reserved
      PubDate: 2015-06-08T09:16:51.910134-05:
      DOI: 10.1002/jcp.25070
       
  • Different Effects of p52SHC1 and p52SHC3 on the Cell Cycle of Neurons and
           Neural Stem Cells
    • Authors: Ning Tang; Dan Lyu, Tao Liu, Fangjin Chen, Shuqian Jing, Tianyu Hao, Shaojun Liu
      Abstract: SHC3 is exclusively expressed in postmitotic neurons, while SHC1 is found in neural stem cells and neural precursor cells but absent in mature neurons. In this study, we discovered that suppression of p52SHC1 expression by RNA interference resulted in proliferation defects in neural stem cells, along with significantly reduced protein levels of cyclin E and cyclin A. At the same time, p52SHC3 RNAi caused cell cycle re‐entry (9.54% in S phase and 5.70% in G2‐M phase) in primary neurons with significantly up‐regulated expression of cyclin D1, cyclin E, cyclin A, CDK2 and phosphorylated CDK2. When p52SHC3 was overexpressed, the cell cycle of neural stem cells was arrested with reduced protein levels of cyclin D1, cyclin E and cyclin A, while overexpression of p52SHC1 did not result in significant changes in postmitotic neurons. Our results indicate that p52SHC3 plays an important role in maintaining the mitotic quiescence of neurons, while p52SHC1 regulates the proliferation of neural stem cells. This article is protected by copyright. All rights reserved
      PubDate: 2015-06-08T09:09:34.439338-05:
      DOI: 10.1002/jcp.25069
       
  • COMMD7 as a novel NEMO interacting protein involved in the termination of
           NF‐κBsignaling
    • Authors: Elio Esposito; Gennaro Napolitano, Alessandra Pescatore, Giuseppe Calculli, Maria Rosaria Incoronato, Antonio Leonardi, Matilde Valeria Ursini
      Abstract: NEMO/IKKγ is the regulatory subunit of the IκB Kinase (IKK) complex, required for the activation of the NF‐κB pathway, which is involved in a variety of key processes, including immunity, inflammation, differentiation and cell survival. Termination of NF‐κB activity on specific ‐κB responsive genes, which is crucial for the resolution of inflammatory responses, can be achieved by direct degradation of the chromatin‐bound NF‐κB subunit RelA/p65, a process mediated by a protein complex that contains Copper Metabolism Murr1 Domain 1 (COMMD1). In this study, we identify COMMD7, another member of the COMMDs protein family, as a novel NEMO‐interacting protein. We show that COMMD7 exerts an inhibitory effect on NF‐κB activation upon TNFα stimulation. COMMD7 interacts with COMMD1 and together they cooperate to down‐regulate NF‐κB activity. Accordingly, termination of TNFα‐induced NF‐κB activity on the ‐κB responsive gene, Icam1, is defective in cells silenced for COMMD7 expression. Furthermore, this impairment is not greatly increased when we silence the expression of both COMMD7 and COMMD1 indicating that the two proteins participate in the same pathway of termination of TNFα‐induced NF‐κB activity. Importantly, we have demonstrated that COMMD7's binding to NEMO does not interfere with the binding to the IKKs, and that the disruption of the IKK complex through the use of the NBP competitor impairs the termination of NF‐κB activity. We propose that an intact IKK complex is required for the termination of NF‐κB‐dependent transcription and that COMMD7 acts as a scaffold in the IKK‐mediated NF‐κB termination. This article is protected by copyright. All rights reserved
      PubDate: 2015-06-08T09:09:11.40997-05:0
      DOI: 10.1002/jcp.25066
       
  • Potential for Stem Cell‐Based Periodontal Therapy
    • Authors: Seyed Hossein Bassir; Wichaya Wisitrasameewong, Justin Raanan, Sasan Ghaffarigarakani, Jamie Chung, Marcelo Freire, Luciano C. Andrada, Giuseppe Intini
      Abstract: Periodontal diseases are highly prevalent and are linked to several systemic diseases.The goal of periodontal treatment is to halt the progression of the disease and regenerate the damaged tissue. However, achieving complete and functional periodontal regeneration is challenging because the periodontium is a complex apparatus composed of different tissues, including bone, cementum, and periodontal ligament. Stem cell‐based regenerative therapy may represent an effective therapeutic tool for periodontal regeneration due to their plasticity and ability to differentiate into different cell lineages. This review presents and critically analyzesthe available information on stem cell‐based therapyfor the regeneration of periodontal tissues and suggests new avenues for the development of more effective therapeutic protocols. This article is protected by copyright. All rights reserved
      PubDate: 2015-06-08T09:08:37.852821-05:
      DOI: 10.1002/jcp.25067
       
  • The Association between p38 MAPK‐Mediated TNF‐α/TNFR2
           Up‐Regulation and
           2‐(4‐Aminophenyl)‐7‐Methoxybenzothiazole‐Induced
           Apoptosis in Human Leukemia U937 Cells
    • Authors: Chia‐Hui Huang; Ying‐Jung Chen, Tzu‐Yu Chao, Wen‐Hsin Liu, Jung‐Jung Changchien, Wan‐Ping Hu, Long‐Sen Chang
      Abstract: The primary cause of treatment failures in acute myeloid leukemia is usually associated with defects in the apoptotic pathway. Several studies suggest that 2‐(4‐aminophenyl)‐7‐methoxybenzothiazole (7‐OMe‐APBT) may potentially induce apoptosis of cancer cells. Thus, the present study was conducted to explore the cytotoxic effect of 7‐OMe‐APBT on human leukemia U937 cells. The apoptosis of human leukemia U937 cells induced by 7‐OMe‐APBT was characterized by an increase in mitochondrial membrane depolarization, procaspase‐8 degradation, and tBid production. Down‐regulation of FADD blocked 7‐OMe‐APBT‐induced procaspase‐8 degradation and rescued the viability of 7‐OMe‐APBT‐treated cells, suggesting the involvement of a death receptor‐mediated pathway in 7‐OMe‐APBT‐induced cell death. Increased TNF‐α expression, TNFR2 expression, and p38 MAPK phosphorylation were noted in 7‐OMe‐APBT‐treated cells. Pretreatment with a p38 MAPK inhibitor abolished 7‐OMe‐APBT‐induced TNF‐α and TNFR2 up‐regulation. 7‐OMe‐APBT stimulated p38 MAPK/c‐Jun‐mediated transcriptional up‐regulation of TNFR2, while the increased TNF‐α mRNA stability led to TNF‐α up‐regulation in 7‐OMe‐APBT‐treated cells. Treatment with 7‐OMe‐APBT up‐regulated protein phosphatase 2A catalytic subunit α (PP2Acα) expression via the p38 MAPK/c‐Jun/ATF‐2 pathway, which, in turn, promoted tristetraprolin (TTP) degradation. Pretreatment with a protein phosphatase 2A inhibitor or TTP over‐expression abrogated TNF‐α up‐regulation in 7‐OMe‐APBT‐treated cells. Abolishment of TNF‐α up‐regulation or knock‐down of TNFR1/TNFR2 by siRNA restored the viability of 7‐OMe‐APBT‐treated cells. Taken together, our data indicate a connection between p38 MAPK‐mediated TNF‐α and TNFR2 up‐regulation and 7‐OMe‐APBT‐induced TNF‐α‐mediated death pathway activation in U937 cells. The same pathway also elucidates the mechanism underlying 7‐OMe‐APBT‐induced death of human leukemia HL‐60 cells. This article is protected by copyright. All rights reserved
      PubDate: 2015-06-08T09:01:09.859542-05:
      DOI: 10.1002/jcp.25064
       
  • Lanthanum Chloride Attenuates Osteoclast Formation and Function via the
           Downregulation of RANKL‐Induced NF‐κB and NFATc1
           Activities
    • Authors: Chuan Jiang; Jiangyinzi Shang, Zhe Li, An Qin, Zhengxiao Ouyang, Xinhua Qu, Haowei Li, Bo Tian, Wengang Wang, Chuanlong Wu, Jinwu Wang, Min Dai
      Abstract: The biological activities of lanthanum chloride (LaCl3) and the molecular mechanisms of action underlying its anti‐inflammatory, anti‐hyperphosphatemic, and osteoblast‐enhancing effects have been studied previously, but less is known about the effects of LaCl3 on osteoclasts. The present study used in vivo and in vitro approaches to explore the effects of LaCl3 on osteoclasts and osteolysis. The results indicated that LaCl3 concentrations that were non‐cytotoxic to mouse bone marrow‐derived monocytes attenuated receptor activator of nuclear factor‐κB ligand (RANKL)‐stimulated osteoclastogenesis, bone resorption, mRNA expression of osteoclastogenic genes in these cells, including cathepsin K, calcitonin receptor, and tartrate‐resistant acid phosphatase (TRAP). Further, LaCl3inhibited RANKL‐mediated activation of the nuclear factor‐κB (NF‐κB) signalingpathway, and downregulated mRNA and protein levels of nuclear factor of activated T‐cells, cytoplasmic, calcineurin‐dependent 1 (NFATc1) and c‐fos. In vivo, LaCl3attenuated titanium (Ti) particle‐induced bone loss in a murine calvarial osteolysis model. Histological analyses revealed that LaCl3 ameliorated bone destruction and decreased the number of TRAP‐positive osteoclasts in this model. These results demonstrated that LaCl3 inhibitedosteoclast formation, function, and osteoclast‐specific gene expression in vitro, and attenuated Ti particle‐induced mouse calvarial osteolysis in vivo, where the inhibition of NF‐κB signaling and downregulation of NFATc1 and c‐fos played an important role. This article is protected by copyright. All rights reserved
      PubDate: 2015-06-08T09:00:26.107893-05:
      DOI: 10.1002/jcp.25065
       
  • C‐ing the Genome: A Compendium of Chromosome Conformation Capture
           Methods to Study Higher‐Order Chromatin Organization
    • Authors: A. Rasim Barutcu; Andrew J. Fritz, Sayyed K. Zaidi, André J. vanWijnen, Jane B. Lian, Janet L. Stein, Jeffrey A. Nickerson, Anthony N. Imbalzano, Gary S. Stein
      Abstract: Three‐dimensional organization of the chromatin has important roles in transcription, replication, DNA repair, and pathologic events such as translocations. There are two fundamental ways to study higher‐order chromatin organization: microscopic and molecular approaches. In this review, we briefly introduce the molecular approaches, focusing on chromosome conformation capture or“3C” technology and its derivatives, which can be used to probe chromatin folding at resolutions beyond that provided by microscopy techniques. We further discuss the different types of data generated by the 3C‐based methods and how they can be used to answer distinct biological questions. This article is protected by copyright. All rights reserved
      PubDate: 2015-06-08T09:00:16.939204-05:
      DOI: 10.1002/jcp.25062
       
  • Mechanisms and Consequences of Double‐strand DNA Break Formation in
           Chromatin
    • Authors: Wendy J. Cannan ; David S. Pederson
      Abstract: All organisms suffer double‐strand breaks (DSBs) in their DNA as a result of exposure to ionizing radiation. DSBs can also form when replication forks encounter certain DNA lesions or repair intermediates. The processing and repair of DSBs can lead to mutations, loss of heterozygosity, and chromosome rearrangements that can lead to cell death or cancer. The most common pathway used to repair DSBs in metazoans (non‐homologous DNA end joining) is more commonly mutagenic than the alternative pathway (homologous recombination mediated repair). Thus, factors that influence the choice of pathways used DSB repair can affect an individual's mutation burden and risk of cancer. This review describes radiological, chemical and biological mechanisms that generate DSBs, and discusses the impact of such variables as DSB etiology, cell type, cell cycle, and chromatin structure on the yield, distribution, and processing of DSBs. The final section will focus on nucleosome‐specific mechanisms that influence DSB production, and the possible relationship between higher order chromosome coiling and chromosome shattering (chromothripsis).This article is protected by copyright. All rights reserved
      PubDate: 2015-06-03T06:06:33.191832-05:
      DOI: 10.1002/jcp.25048
       
  • Animal Models for Progressive Multifocal Leukoencephalopathy
    • Authors: Martyn K. White ; Jennifer Gordon, Joseph R. Berger, Kamel Khalili
      Abstract: Progressive multifocal leukoencephalopathy (PML) is a severe demyelinating disease of the CNS caused by the human polyomavirus JC (JCV). JCV replication occurs only in human cells and investigation of PML has been severely hampered by the lack of an animal model. The common feature of PML is impairment of the immune system. The key to understanding PML is working out the complex mechanisms that underlie viral entry and replication within the CNS and the immunosurveillance that suppresses the virus or allows it to reactivate. Early models involved the simple inoculation of JCV into animals such as monkeys, hamsters, and mice. More recently, mouse models transgenic for the gene encoding the JCV early protein, T‐antigen, a protein thought to be involved in the disruption of myelin seen in PML, have been employed. These animal models resulted in tumorigenesis rather than demyelination. Another approach is to use animal polyomaviruses that are closely related to JCV but able to replicate in the animal such as mouse polyomavirus and SV40. More recently, novel models have been developed that involve the engraftment of human cells into the animal. Here, we review progress that has been made to establish an animal model for PML, the advances and limitations of different models and weigh future prospects.This article is protected by copyright. All rights reserved
      PubDate: 2015-06-03T06:06:06.456436-05:
      DOI: 10.1002/jcp.25047
       
  • Brg1 Controls the Expression of Pax7 to Promote Viability and
           Proliferation of Mouse Primary Myoblasts
    • Authors: Teresita Padilla‐Benavides ; Brian T. Nasipak, Anthony N. Imbalzano
      Abstract: Brg1 (Brahma‐related gene 1) is a catalytic component of the evolutionarily conserved mammalian SWI/SNF ATP‐dependent chromatin remodeling enzymes that disrupt histone‐DNA contacts on the nucleosome. While the requirement for the SWI/SNF enzymes in cell differentiation has been extensively studied, its role in precursor cell proliferation and survival is not as well defined. Muscle satellite cells constitute the stem cell pool that sustains and regenerates myofibers in adult skeletal muscle. Here, we show that deletion of Brg1 in primary mouse myoblasts derived from muscle satellite cells cultured ex vivo leads to a cell proliferation defect and apoptosis. We determined that Brg1 regulates cell proliferation and survival by controlling chromatin remodeling and activating transcription at the Pax7 promoter, which is expressed during somite development and is required for controlling viability of the satellite cell population. Reintroduction of catalytically active Brg1 or of Pax7 into Brg1‐deficient satellite cells rescued the apoptotic phenotype and restored proliferation. These data demonstrate that Brg1 functions as a positive regulator for cellular proliferation and survival of primary myoblasts. Therefore, the regulation of gene expression through Brg1‐mediated chromatin remodeling is critical not just for skeletal muscle differentiation but for maintaining the myoblast population as well. This article is protected by copyright. All rights reserved
      PubDate: 2015-06-02T08:35:40.093852-05:
      DOI: 10.1002/jcp.25031
       
  • Establishment of Immortalized Mouse Bmp2 Knock‐out Dental Papilla
           Mesenchymal Cells Necessary for Study of Odontoblastic Differentiation and
           Odontogenesis
    • Authors: Lian Wu; Feng Wang, Kevin J. Donly, Chunyan Wan, Daoshu Luo, Stephen E. Harris, Mary MacDougall, Shuo Chen
      Abstract: Bmp2 is essential for dentin formation. Bmp2 cKO mice exhibited similar phenotype to dentinogenesis imperfecta, showing dental pulp exposure, hypomineralized dentin and delayed odontoblast differentiation. As it is relatively difficult to obtain lot of primary Bmp2 cKO dental papilla mesenchymal cells and to maintain a long term culture of these primary cells, Availability of immortalized deleted Bmp2 dental papilla mesenchymal cells is critical for studying the underlying mechanism of Bmp2 signal in odontogenesis. In this study, our goal was to generate an immortalized deleted Bmp2 dental papilla mesenchymal (iBmp2ko/ko dp) cell line by introducing Cre recombinase and green fluorescent protein (FGP) into the immortalized mouse floxed Bmp2 dental papilla mesenchymal (iBmp2fx/fx dp) cells. iBmp2ko/ko dp cells were confirmed by GFP and PCR. The deleted Bmp2 cells exhibited slow cell proliferation rate and cell growth was arrested in G2 phase. Expression of tooth‐related marker genes and cell differentiation were decreased in the deleted cells. Importantly, extracellular matrix remodeling was impaired in the iBmp2ko/ko dp cells as reflected by the decreased Mmp‐9 expression. In addition, with exogenous Bmp2 induction, these cell differentiation and mineralization were rescued as well as extracellular matrix remodeling was enhanced. Therefore, we for the first time described establishment of iBmpko/ko cells that are useful for study of mechanisms in regulating dental papilla mesenchymal cell lineages. This article is protected by copyright. All rights reserved
      PubDate: 2015-06-02T07:45:59.458148-05:
      DOI: 10.1002/jcp.25061
       
  • EGF‐Induced Connexin43 Negatively Regulates Cell Proliferation in
           Human Ovarian Cancer
    • Authors: Xin Qiu; Jung‐Chien Cheng, Christian Klausen, Hsun‐Ming Chang, Qianlan Fan, Peter C.K. Leung
      Abstract: Connexin43 (Cx43) has been shown to regulate cell proliferation and its down‐regulation is correlated with poor prognosis and survival in several types of human cancer. Cx43 expression levels are frequently down‐regulated in human ovarian cancer, suggesting a potential role for Cx43 in regulating the progression of this disease. Epidermal growth factor (EGF) is a well‐characterized hormone that stimulates ovarian cancer cell proliferation. Although EGF is able to regulate Cx43 expression in other cell types, it is unclear whether EGF can regulate Cx43 expression in ovarian cancer cells. Additionally, it remains unknown whether Cx43 is involved in EGF‐stimulated ovarian cancer cell proliferation. In the present study, we demonstrate that treatment with EGF up‐regulates Cx43 expression in two ovarian cancer cell lines, SKOV3 and OVCAR4. Although treatment with EGF activates both ERK1/2 and Akt signaling pathways, pharmacological inhibition and siRNA‐mediated knockdown suggest that only the activation of Akt1 is required for EGF‐induced Cx43 up‐regulation. Functionally, Cx43 knockdown enhanced basal and EGF‐induced cell proliferation, whereas the proliferative effects of EGF were reduced by Cx43 overexpression. Co‐treatment with the gap junction inhibitor carbenoxolone did not alter the suppressive effects of Cx43 overexpression on EGF‐induced cell proliferation, suggesting a gap junction‐independent mechanism. This study reveals an important role for Cx43 as a negative regulator of EGF‐induced human ovarian cancer cell proliferation. This article is protected by copyright. All rights reserved
      PubDate: 2015-05-29T18:44:55.234757-05:
      DOI: 10.1002/jcp.25058
       
  • Cell‐Penetrating and Endoplasmic Reticulum‐Locating
           TAT‐IL‐24‐KDEL Fusion Protein Induces Tumor Apoptosis
    • Authors: Jian Zhang; Aiyou Sun, Rui Xu, Xinyi Tao, Yuguo Dong, Xinxin Lv, Dongzhi Wei
      Abstract: Interleukin‐24 (IL‐24) is a unique IL‐10 family cytokine that could selectively induce apoptosis in cancer cells without harming normal cells. Previous research demonstrated that intracellular IL‐24 protein induces an endoplasmic reticulum (ER) stress response only in cancer cells, culminating in apoptosis. In this study, we developed a novel recombinant fusion protein to penetrate into cancer cells and locate on ER. It is composed of three distinct functional domains, IL‐24, and the targeting domain of transactivator of transcription (TAT) and an ER retention four‐peptide sequence KDEL (Lys‐Asp‐Glu‐Leu) that link at its NH2 and COOH terminal, respectively. The in vitro results indicated that TAT‐IL‐24‐KDEL inhibited growth in bladder cancer cells, as well as in non‐small cell lung cancer cell line and breast cancer cell line, but the normal human lung fibroblast cell line was not affected, indicating the cancer specificity of TAT‐IL‐24‐KDEL. Western blot analysis showed that apoptosis activation was induced by TAT‐IL‐24‐KDEL through the ER stress‐mediated cell death pathway. Treatment with TAT‐IL‐24‐KDEL significantly inhibited the growth of human H460 xenografts in nude mice, and the tumor growth inhibition was correlated with increased hematoxylin and eosin (H&E) staining and terminal deoxynucleotidyl transferase‐mediated dUTP nick end labeling (TUNEL) staining. These findings suggest that the artificially designed recombinant fusion protein TAT‐IL‐24‐KDEL may be highly effective in cancer therapy and worthy of further evaluation and development. This article is protected by copyright. All rights reserved
      PubDate: 2015-05-29T18:44:35.492967-05:
      DOI: 10.1002/jcp.25054
       
  • Error‐Prone Repair of DNA Double‐Strand Breaks
    • Authors: Kasey Rodgers; Mitch McVey
      Abstract: Preserving the integrity of the DNA double helix is crucial for the maintenance of genomic stability. Therefore, DNA double‐strand breaks represent a serious threat to cells. In this review, we describe the two major strategies used to repair double strand breaks: non‐homologous end joining and homologous recombination, emphasizing the mutagenic aspects of each. We focus on emerging evidence that homologous recombination, long thought to be an error‐free repair process, can in fact be highly mutagenic, particularly in contexts requiring large amounts of DNA synthesis. Recent investigations have begun to illuminate the molecular mechanisms by which error‐prone double‐strand break repair can create major genomic changes, such as translocations and complex chromosome rearrangements. We highlight these studies and discuss proposed models that may explain some of the more extreme genetic changes observed in human cancers and congenital disorders. This article is protected by copyright. All rights reserved
      PubDate: 2015-05-29T18:44:16.245325-05:
      DOI: 10.1002/jcp.25053
       
  • p53 Modulates Notch Signaling in MCF‐7 Breast Cancer Cells by
           Associating with the Notch Transcriptional Complex via MAML1
    • Authors: Jieun Yun; Ingrid Espinoza, Antonio Pannuti, Damian Romero, Luis Martinez, Mary Caskey, Adina Stanculescu, Maurizio Bocchetta, Paola Rizzo, Vimla Band, Hamid Band, Hwan Mook Kim, Song‐Kyu Park, Keon Wook Kang, Maria Laura Avantaggiati, Christian R. Gomez, Todd Golde, Barbara Osborne, Lucio Miele
      Abstract: p53 and Notch‐1 play important roles in breast cancer biology. Notch‐1 inhibits p53 activity in cervical and breast cancer cells. Conversely, p53 inhibits Notch activity in T‐cells but stimulates it in human keratinocytes. Notch co‐activator MAML1 binds p53 and functions as a p53 co‐activator. We studied the regulation of Notch signaling by p53 in MCF‐7 cells and normal human mammary epithelial cells (HMEC). Results show that overexpression of p53 or activation of endogenous p53 with Nutlin‐3 inhibits Notch‐dependent transcriptional activity and Notch target expression in a dose‐dependent manner. This effect could be partially rescued by transfection of MAML1 but not p300. Standard and quantitative co‐immunoprecipitation experiments readily detected a complex containing p53 and Notch‐1 in MCF‐7 cells. Formation of this complex was inhibited by dominant negative MAML1 (DN‐MAML1) and stimulated by wild‐type MAML1. Standard and quantitative far‐Western experiments showed a complex including p53, Notch‐1 and MAML1. Chromatin immunoprecipitation (ChIP) experiments showed that p53 can associate with Notch‐dependent HEY1 promoter and this association is inhibited by DN‐MAML1 and stimulated by wild‐type MAML1. Our data support a model in which p53 associates with the Notch transcriptional complex (NTC) in a MAML1‐dependent fashion, most likely through a p53‐MAML1 interaction. In our cellular models, the effect of this association is to inhibit Notch‐dependent transcription. Our data suggest that p53‐null breast cancers may lack this Notch‐modulatory mechanism, and that therapeutic strategies that activate wild‐type p53 can indirectly cause inhibition of Notch transcriptional activity. This article is protected by copyright. All rights reserved
      PubDate: 2015-05-29T18:43:59.77372-05:0
      DOI: 10.1002/jcp.25052
       
  • microRNA as Biomarkers and Diagnostics
    • Authors: Jin Wang; Jinyun Chen, Subrata Sen
      Abstract: MicroRNAs (miRNAs) are a group of small non‐coding RNAs that are involved in regulating a range of developmental and physiological processes; their dysregulation has been associated with development of diseases including cancer. Circulating miRNAs and exosomal miRNAs have also been proposed as being useful in diagnostics as biomarkers for diseases and different types of cancer. In this review, miRNAs are discussed as biomarkers for cancer and other diseases, including viral infections, nervous system disorders, cardiovascular disorders, and diabetes. We summarize some of the clinical evidence for the use of miRNAs as biomarkers in diagnostics and provide some general perspectives on their use in clinical situations. The analytical challenges in using miRNAs in cancer and disease diagnostics are evaluated and discussed. Validation of specific miRNA signatures as biomarkers is a critical milestone in diagnostics. This article is protected by copyright. All rights reserved
      PubDate: 2015-05-29T18:43:44.603437-05:
      DOI: 10.1002/jcp.25056
       
  • Endotoxin‐Stimulated Rat Hepatic Stellate Cells Induce Autophagy in
           Hepatocytes as a Survival Mechanism
    • Authors: Anil Dangi; Chao Huang, Ashish Tandon, Donna Stolz, Tong Wu, Chandrashekhar R. Gandhi
      Abstract: Bacterial lipopolysaccharide (LPS)‐stimulated hepatic stellate cells (HSCs) produce many cytokines including IFNβ, TNFα and IL6, strongly inhibit DNA synthesis, but induce apoptosis of a small number of hepatocytes. In vivo administration of LPS (up to 10 mg/ml) causes modest inflammation and weight loss in rats but not mortality. We determined whether LPS‐stimulated HSCs instigate mechanisms of hepatocyte survival. Rats received 10 mg/kg LPS (i.p.) and determinations were made at 6h. In vitro, HSCs were treated with 100 ng/ml LPS till 24h. The medium was transferred to hepatocytes, and determinations were made at 0‐12h. Controls were HSC‐conditioned medium or medium‐containing LPS. LPS treatment of rats caused autophagy in hepatocytes, a physiological process for clearance of undesirable material including injured or damaged organelles. This was accompanied by activation of c‐Jun NH2 terminal kinase (JNK) and apoptosis of ∼4‐5% of hepatocytes. In vitro, LPS‐conditioned HSC medium (LPS/HSC) induced autophagy in hepatocytes but apoptosis of only ∼10% of hepatocytes. While LPS/HSC stimulated activation of JNK (associated with cell death), it also activated NFkB and ERK1/2 (associated with cell survival). LPS‐stimulated HSCs produced IFNβ, and LPS/HSC‐induced autophagy in hepatocytes and their apoptosis were significantly inhibited by anti‐IFNβ antibody. Blockade of autophagy, on the other hand, strongly augmented hepatocyte apoptosis. While LPS‐stimulated HSCs cause apoptosis of a subpopulation of hepatocytes by producing IFNβ, they also induce cell survival mechanisms, which may be of critical importance in resistance to liver injury during endotoxemia. This article is protected by copyright. All rights reserved
      PubDate: 2015-05-29T18:43:27.71238-05:0
      DOI: 10.1002/jcp.25055
       
  • Enhanced Chemotherapeutic Behavior of Open‐Caged DNA@Doxorubicin
           Nanostructures for Cancer Cells
    • Authors: Vinit Kumar; Samer Bayda, Mohamad Hadla, Isabella Caligiuri, Concetta Russo Spena, Stefano Palazzolo, Susanne Kempter, Giuseppe Corona, Giuseppe Toffoli, Flavio Rizzolio
      Abstract: In cancer therapy, it is imperative to increase the efficacy and reduce side effects of chemotherapeutic drugs. Nanotechnology offers the unique opportunity to overcome these barriers. In particular, in the last few years DNA nanostructures have gained attention for their biocompatibility, easy customized synthesis and ability to deliver drugs to cancer cells. Here, an open‐caged pyramidal DNA@Doxorubicin (Py‐Doxo) nanostructure was constructed with 10 DNA sequences of 26–28 nucleotides for drug delivery to cancer cells. The synthesized DNA nanostructures are sufficiently stable in biological medium. Py‐Doxo exhibited significantly enhanced cytotoxicity of the delivered doxorubicin to breast and liver cancer cells up to two fold compared to free doxorubicin. This study demonstrates the importance of the shape and structure of the designed transporter DNA nanostructures for biomedical applications. This article is protected by copyright. All rights reserved
      PubDate: 2015-05-29T18:43:10.761567-05:
      DOI: 10.1002/jcp.25057
       
  • MicroRNAs Provide Feedback Regulation of Epithelial‐Mesenchymal
           Transition Induced by Growth Factors
    • Authors: Zailong Qin; Wei He, Jingqun Tang, Qiurong Ye, Wei Dang, Yuanjun Lu, Jia Wang, Guiyuan Li, Qun Yan, Jian Ma
      Abstract: As regulators in gene expression, microRNAs take part in most biological processes including cell differentiation, apoptosis, cell cycle and epithelial‐to‐mesenchymal transition (EMT). In order to evaluate their roles in EMT process, microRNA expression profile changes induced by EGF or TGF‐β treatment on nasopharyngeal carcinoma cell HK‐1 were analyzed, and miR‐21, miR‐148a, miR‐505 and miR‐1207‐5p were found to be upregulated in growth factors‐induced EMT process. miR‐21 is already known as an oncogenic miRNA to promote metastasis, however, the exact functions of other three miRNAs in EMT are unclear. To our surprise, we found that miR‐148a, miR‐505 and miR‐1207‐5p can suppress EMT and metastasis phenotypes in HK‐1 cells both in vitro and in vivo, which may relate to their inhibition on EMT and Wnt signaling molecules. MiRNAs confer robustness to biological processes by posttranscriptional repression of key transcriptional programs that are related to previous developmental stages or to alternative cell fates. Our findings indicate that miRNA feedback circuit is tuned to respond to growth factors‐induced EMT, and we suggested a new negative feedback loop which may be an important element of the EMT process and confer biological robustness. This article is protected by copyright. All rights reserved
      PubDate: 2015-05-29T18:42:52.178496-05:
      DOI: 10.1002/jcp.25060
       
  • Bioregulation
    • Authors: Arthur B. Pardee
      Abstract: Regulation is at the heart of biology. What bioregulatory mechanisms finely tune and coordinate our cell's numerous complicated biochemical processes, maintaining constancy of the cell internal environment when external conditions change and it differentiates during development? Knowledge of the biochemistry of small molecules in normal cells progressed greatly in the early 1900s. This article is protected by copyright. All rights reserved
      PubDate: 2015-05-29T18:42:35.629388-05:
      DOI: 10.1002/jcp.25059
       
  • Acidosis Is a Key Regulatior of Osteoblast Ecto‐Nucleotidase
           Pyrophosphatase/Phosphodiesterase 1 (NPP1) Expression and Activity
    • Authors: Isabel R Orriss; Michelle L Key, Mark OR Hajjawi, José Luis Millán, Timothy R Arnett
      Abstract: Previous work has shown that acidosis prevents bone nodule formation by osteoblasts in vitro by inhibiting mineralisation of the collagenous matrix. The ratio of phosphate (Pi) to pyrophosphate (PPi) in the bone microenvironment is a fundamental regulator of bone mineralisation. Both Pi and PPi, a potent inhibitor of mineralisation, are generated from extracellular nucleotides by the actions of ecto‐nucleotidases. This study investigated the expression and activity of ecto‐nucleotidases by osteoblasts under normal and acid conditions. We found that osteoblasts express mRNA for a number of ecto‐nucleotidases including NTPdase 1‐6 (ecto‐nucleoside triphosphate diphosphohydrolase) and NPP1‐3 (ecto‐nucleotide pyrophosphatase/phosphodiesterase). The rank order of mRNA expression in differentiating rat osteoblasts (day 7) was Enpp1 > NTPdase 4 > NTPdase 6 > NTPdase 5 > alkaline phosphatase > ecto‐5‐nucleotidase > Enpp3 > NTPdase 1 > NTPdase 3 > Enpp2 > NTPdase 2. Acidosis (pH 6.9) upregulated NPP1 mRNA (2.8‐fold) and protein expression at all stages of osteoblast differentiation compared to physiological pH (pH 7.4); expression of other ecto‐nucleotidases was unaffected. Furthermore, total NPP activity was increased up to 53% in osteoblasts cultured in acid conditions (p 
      PubDate: 2015-05-29T18:31:24.934646-05:
      DOI: 10.1002/jcp.25041
       
  • Stable Incretin Mimetics Counter Rapid Deterioration of Bone Quality in
           Type 1 Diabetes Mellitus
    • Authors: Sity Aishah Mansur; Aleksandra Mieczkowska, Béatrice Bouvard, Peter R Flatt, Daniel Chappard, Nigel Irwin, Guillaume Mabilleau
      Abstract: Aims Type 1 diabetes mellitus is associated with a high risk for bone fractures. Although bone mass is reduced, bone quality is also dramatically altered in this disorder. However, recent evidences suggest a beneficial effect of the glucose‐dependent insulinotropic polypeptide (GIP) and glucagon‐like peptide 1 (GLP‐1) pathways on bone quality. The aims of the present study were to conduct a comprehensive investigation of bone strength at the organ and tissue level; and to ascertain whether enzyme resistant GIP or GLP‐1 mimetic could be beneficial in preventing bone fragility in type 1 diabetes mellitus. Materials and methods Streptozotocin‐treated mice were used as a model of type 1 diabetes mellitus. Control and streptozotocin‐diabetic animals were treated for 21 days with an enzymatic‐resistant GIP peptide ([D‐Ala2]GIP) or with liraglutide (each at 25 nmol/kg bw, ip). Bone quality was assessed at the organ and tissue level by microCT, qXRI, 3‐point bending, qBEI, nanoindentation and Fourier‐transform infrared microspectroscopy. Results [D‐Ala2]GIP and liraglutide treatment did prevent loss of whole bone strength and cortical microstructure in the STZ‐injected mice. However, tissue material properties were significantly improved in STZ‐injected animals following treatment with [D‐Ala2]GIP or liraglutide. Conclusions Treatment of STZ‐diabetic mice with [D‐Ala2]GIP or liraglutide was capable of significantly preventing deterioration of the quality of the bone matrix. Further studies are required to further elucidate the molecular mechanisms involved and to validate whether these findings can be translated to human patients. This article is protected by copyright. All rights reserved
      PubDate: 2015-05-27T12:56:27.572693-05:
      DOI: 10.1002/jcp.25033
       
  • The Effects of miR‐20a on p21: Two Mechanisms Blocking Growth Arrest
           in TGF‐β Responsive Colon Carcinoma
    • Authors: Viktorija Sokolova; Antonio Fiorino, Eugenio Zoni, Elisabetta Crippa, James F Reid, Manuela Gariboldi, Marco A Pierotti
      Abstract: Loss of response to TGF‐β is a central event in the genesis of colorectal cancer (CRC), a disease that, in the majority cases, is refractory to growth inhibition induced by this cytokine. However, inactivating mutations at receptors and transducers from the TGF‐β cascade occur only in approximately half of CRCs, suggesting the involvement of additional mechanisms altering the response to the cytokine. We have recently described the amplification of the 13q31 locus, where the miR‐17‐92 cluster maps, associated with overexpression of members of the cluster. In this study, we address the potential role of miR‐20a, from the miR‐17‐92 cluster, in the suppression of TGF‐β cytostatic response in CRC. Using the poorly tumorigenic and TGF‐β sensitive FET cell line that expresses low miR‐20a levels, we first confirmed that miR‐20a down‐modulated CDKN1A expression, both at mRNA and protein level, through direct binding to its 3'‐UTR. We demonstrated that miR‐20a significantly diminished cell response to TGF‐β by preventing its delay of G1/S transition and promoting progression into cell cycle. Moreover, besides modulating CDKN1A, miR‐20a blocked TGF‐β‐induced transactivation of its promoter without affecting the post‐receptor activation of Smad3/4 effectors. Finally, miR‐20a abrogated the TGF‐β mediated c‐Myc repression, a direct inhibitor of the CDKN1A promoter activation, most likely by reducing the expression of specific MYC‐regulating genes from the Smad/E2F‐based core repressor complex. Collectively, our experiments indicate that miR‐20a interferes with the colonic epithelium homeostasis by disrupting the regulation of Myc/p21 by TGF‐β, which is essential for its malignant transformation. This article is protected by copyright. All rights reserved
      PubDate: 2015-05-26T17:52:50.46159-05:0
      DOI: 10.1002/jcp.25051
       
  • Dual Effect of Adenosine A1 Receptor Activation on Renal O2 Consumption
    • Authors: Victor Babich; Komal Vadnagara, Francesca Di Sole
      Abstract: The high requirement of O2 in the renal proximal tubule stems from a high rate of Na+ transport. Adenosine A1 receptor (A1R) activation regulates Na+ transport in this nephron segment. Thus, the effect of the acute activation and the mechanisms of A1R on the rate of O2 consumption were evaluated. The A1R‐antagonist, 8‐cyclopentyl‐1,3‐dipropylxanthine (CPX) and adenosine deaminase (ADA), which metabolize endogenous adenosine, reduced O2 consumption (40‐50%). Replacing Na+ in the buffer reversed the ADA‐ or CPX‐mediated reduction of O2 consumption. Blocking the Na/H‐exchanger activity, which decreases O2 usage per se, did not enhance the ADA‐ or CPX‐induced inhibition of O2 consumption. These data indicate that endogenous adenosine increases O2 usage via the activation of Na+ transport. In the presence of endogenous adenosine, A1R was further activated by the A1R‐agonist N6‐cyclopentyladenosine (CPA); CPA inhibited O2 usage (30%) and this effect also depended on Na+ transport. Moreover, a low concentration of CPA activated O2 usage in tissue pretreated with ADA, whereas a high concentration of CPA inhibited O2 usage; both effects depended on Na+. Protein kinase C signaling mediated the inhibitory effect of A1R, while adenylyl cyclase mediated its stimulatory effect on O2 consumption. In summary, increasing the local concentrations of adenosine can either activate or inhibit O2 consumption via A1R, and this mechanism depends on Na+ transport. The inhibition of O2 usage by A1R activation might restore the compromised balance between energy supply and demand under pathophysiological conditions, such as renal ischemia, which results in high adenosine production. This article is protected by copyright. All rights reserved
      PubDate: 2015-05-23T17:47:44.209202-05:
      DOI: 10.1002/jcp.25050
       
  • Differential Hematopoietic Activity in White Adipose Tissue Depending on
           its Localization
    • Authors: Elodie Luche; Coralie Sengenès, Emmanuelle Arnaud, Patrick Laharrague, Louis Casteilla, Beatrice Cousin
      Abstract: White adipose tissue (WAT) can be found in different locations in the body, and these different adipose deposits exhibit specific physiopathological importance according to the subcutaneous or abdominal locations. We have shown previously the presence of functional hematopoietic stem/progenitor cells (HSPC) in subcutaneous adipose tissue (SCAT). These cells exhibit a specific hematopoietic activity that contributes to the renewal of the immune cell compartment within this adipose deposit. In this study we investigated whether HSPC can be found in visceral adipose tissue (VAT) and whether a putative difference in in situ hematopoiesis may be related to anatomical location and to site‐specific immune cell content in VAT compared to SCAT. Therein, we identified for the first time the presence of HSPC in VAT. Using both in vitro assays and in vivo competitive repopulation experiments with sorted HSPC from VAT or SCAT, we showed that the hematopoietic activity of HSPC was lower in VAT, compared to SCAT. In addition, this altered hematopoietic activity of HSPC in VAT was due to their microenvironment, and may be related to a specific combination of secreted factors and extracellular matrix molecules expressed by adipose derived stromal cells. Our results indicate that WAT specific hematopoietic activity may be generalized to all adipose deposits, although with specificity according to the fat pad location. Considering the abundance of WAT in the body, this emphasizes the potential importance of this hematopoietic activity in physiopathological situations. This article is protected by copyright. All rights reserved
      PubDate: 2015-05-18T17:51:56.231608-05:
      DOI: 10.1002/jcp.25045
       
  • Involvement of a P2 × 7 Receptor in the Acrosome Reaction
           Induced by ATP in Rat Spermatozoa
    • Authors: Jorge L. Torres‐Fuentes; Mariana Rios, Ricardo D. Moreno
      Abstract: The acrosome reaction (AR) is the exocytosis of the acrosomal vesicle in response to different physiological and non‐physiological stimuli. Particularly in mammals, the AR is needed for sperm to fuse with the oocyte plasma membrane, and it occurs only in capacitated sperm. Previous evidence in the literature indicates that extracellular ATP induces the AR in capacitated human and bovine spermatozoa, but its receptor has not yet been identified. The aim of this work was to define a putative ATP receptor in rat spermatozoa using pharmacological and biochemical approaches. We found that ATP induced the AR only in capacitated rat spermatozoa, which was inhibited in the presence of two general inhibitors of ATP receptors (P2 receptors), Suramin, and oxidized ATP (oATP), and one inhibitor of P2X receptor (pyridoxalphosphate‐6‐azophenyl‐2',4'‐disulfonic acid (PPADS)). In addition, the AR induced by ATP in capacitated rat spermatozoa was inhibited by brilliant blue‐G (BB‐G) and 17‐β‐oestradiol, two blockers of P2 × 7 receptors. Moreover, the ATP analog 2'(3')‐O‐(4‐Benzoylbenzoyl) ATP (BzATP) was almost 500 times more potent than ATP to induce the AR, which agrees with the pharmacology of a P2 × 7 receptor. Here we show the presence of P2 × 7 receptor by Western blot and its localization in the tail and acrosome by indirect immunofluorescence. Finally, we quantify the presence of ATP in the rat oviduct during the estrous cycle. We found that the ATP concentration within the lumen of the oviduct is similar to those required to induce acrosome reaction, which agree with its role during in vivo fertilization. Therefore, our results strongly suggest that ATP induces the AR in capacitated rat spermatozoa through a P2 × 7 receptor, which may be functional during in vivo fertilization. This article is protected by copyright. All rights reserved
      PubDate: 2015-05-18T17:50:53.03108-05:0
      DOI: 10.1002/jcp.25044
       
  • The Many Faces of the A2b Adenosine Receptor in Cardiovascular and
           Metabolic Diseases
    • Authors: Anna Eisenstein; Shenia Patterson, Katya Ravid
      Abstract: Modulation of the low affinity adenosine receptor subtype, the A2b adenosine receptor (A2bAR), has gained interest as a therapeutic target in various pathologic areas associated with cardiovascular disease. The actions of the A2bAR are diverse and at times conflicting depending on cell and tissue type and the timing of activation or inhibition of the receptor. The A2bAR is a promising and exciting pharmacologic target, however, a thorough understanding of A2bAR action is necessary to reach the therapeutic potential of this receptor. This review will focus on the role of the A2bAR in various cardiovascular and metabolic pathologies in which the receptor is currently being studied. We will illustrate the complexities of A2bAR signaling and highlight areas of research with potential for therapeutic development. This article is protected by copyright. All rights reserved
      PubDate: 2015-05-14T18:05:22.807075-05:
      DOI: 10.1002/jcp.25043
       
  • Differential Cellular Effects of Plk1 Inhibitors Targeting the
           ATP‐Binding Domain or Polo‐Box Domain
    • Authors: Sol‐Bi Shin; Sang‐Uk Woo, Hyungshin Yim
      Abstract: The expression of polo‐like kinase 1 (Plk1) correlates with malignancy and is thus recognized as a target for cancer therapy. In addition to the development of ATP‐competitive Plk1 inhibitors, the polo‐box domain (PBD), a unique functional domain of PLKs, is being targeted to develop Plk1‐specific inhibitors. However, the action mechanisms of these two classes of Plk1 inhibitors have not been thoroughly evaluated. Here, we evaluate the differences in cellular effects of ATP‐binding domain inhibitors (BI2536, GSK461364) and PBD inhibitors (poloxin, thymoquinone) to determine their mechanisms of Plk1 inhibition. Our data show that BI2536 and GSK461364 increased the population of cells in the G2/M phase compared with controls, while treatment with poloxin and thymoquinone increased cell population in the S phase as well as in G2/M, in a p53‐independent manner. The population of cells staining positively for p‐Histone H3 and MPM2, mitotic index, was increased by treatment with BI2536 or GSK461364, but not by treatment with poloxin or thymoquinone. Furthermore, treatment with BI2536 or GSK461364 resulted in activation of the BubR1 spindle checkpoint kinase, suggesting that treatment with ATP‐binding domain inhibitors induces metaphase arrest. However, the administration of poloxin and thymoquinone resulted in an increase in p21WAF1 and S arrest, indicating that PBD inhibitors also affected interphase before mitotic entry. Taken together, these data suggest that the PDB of Plk1 plays a role in S phase progression through interaction with other proteins, while its ATP‐binding domain is important for regulating mitotic progression mediated by its catalytic activity involving consumption of ATP. This article is protected by copyright. All rights reserved
      PubDate: 2015-05-14T18:04:58.96938-05:0
      DOI: 10.1002/jcp.25042
       
  • Electrospun Type 1 Collagen Matrices Using a Novel Benign Solvent for
           Cardiac Tissue Engineering
    • Authors: Alan M. Punnoose; Anuradha Elamparithi, Sarah Kuruvilla
      Abstract: Electrospinning is a well‐established technique that uses a high electric field to fabricate ultra fine fibrous scaffolds from both natural and synthetic polymers to mimic the cellular microenvironment. Collagen is one of the most preferred biopolymers due to its biocompatibility and widespread occurrence in nature. Electrospinning of Collagen alone has been reported with fluoroalcohols such as Hexafluoroisopropanol (HFIP) and Trifluoroethanol (TFE), which are toxic to the environment. In this study we describe the use of a novel benign binary solvent to generate nanofibers of Collagen type 1, which is non‐toxic and economical. Transmission electron microscopy (TEM) analysis revealed the characteristic feature of native collagen namely the 67 nm banding pattern, confirming that the triple helical structure was maintained. Scanning Electron Microscopy (SEM) analysis showed the fiber diameters to be in the 200‐800 nm range. Biocompatibility of the three dimensional (3D) scaffolds was established by MTT assays using skeletal myoblasts and Confocal Microscopic analysis of immunofluorescent stained sections for muscle specific markers such as Desmin and Actin. Primary neonatal rat ventricular cardiomyocytes seeded onto the scaffolds were able to maintain their contractile function for a period of 17 days. Our work provides evidence that Collagen 1 can be electrospun without combining with other polymers using a novel benign solvent and we are currently exploring the potential of this approach for cardiac and skeletal muscle tissue engineering. This article is protected by copyright. All rights reserved
      PubDate: 2015-05-12T13:48:51.922438-05:
      DOI: 10.1002/jcp.25035
       
  • Deficiency in the α1 Subunit of Na+/K+‐ATPase Enhances the
           Anti‐Proliferative Effect of High Osmolality in Nucleus Pulposus
           Intervertebral Disc Cells
    • Authors: Eleni Mavrogonatou; Konstantinos Papadimitriou, Jill P. Urban, Vassilios Papadopoulos, Dimitris Kletsas
      Abstract: Intervertebral disc cells are constantly exposed to a hyperosmotic environment. Among cellular responses towards this stress is the inhibition of proliferation through the activation of p38 MAPK and p53. In an effort to further elucidate the biochemical pathways triggered by hyperosmotic stress, we assessed the high osmolality‐induced transcriptional changes of bovine nucleus pulposus cells using whole‐genome arrays. A 5‐ and a 24‐h hyperosmotic treatment led to the differential expression of >100 and >200 genes, respectively, including nine genes encoding transporters (SLC4A11, SLC5A3, ATP1A1, SLC38A2, KCNK17, KCTD20, KCTD11, SLC7A5 and CLCA2). Differences in the transcriptional profile of these selected genes, as indicated by the microarrays experiments, were validated by qRT‐PCR in 2D and 3D cell cultures, under hyperosmolar salt and sorbitol conditions, revealing the presence of a common triggering signal for osmotic adaptation. The key signaling molecules p38 MAPK and p53 were demonstrated to differently participate in the regulation of the aforementioned transporters. Finally, siRNA‐mediated knocking‐down of each one of the three transporters with the highest and sustained over‐expression (i.e. SLC4A11, SLC5A3 and ATP1A1) had a distinct outcome on the transcriptional profile of the other transporters, on p38 MAPK and p53 phosphorylation and consequently on cell cycle progression. The inhibition of ATP1A1 had the most prominent effect on the transcription of the rest of the transporters and was found to enhance the anti‐proliferative effect of hyperosmotic conditions through an increased G2/M cell cycle block, ascribing to this pump a central role in the osmoregulatory response of nucleus pulposus cells. This article is protected by copyright. All rights reserved
      PubDate: 2015-05-12T13:48:07.380527-05:
      DOI: 10.1002/jcp.25040
       
  • Coalition of Nuclear Receptors in the Nervous System
    • Authors: Benjamin Förthmann; John M. Aletta, Yu‐Wei Lee, Chris Terranova, Barbara Birkaya, Ewa K. Stachowiak, Michal K. Stachowiak, Peter Claus
      Abstract: A universal signaling module has been described which utilizes the nuclear form of Fibroblast growth Factor Receptor (FGFR1) in a central role directing the post‐mitotic development of neural cells through coordinated gene expression. In this review, we discuss in detail the current knowledge of FGFR1 nuclear interaction partners in three scenarios: (i) Engagement of FGFR1 in neuronal stem cells and regulation of neuronal differentiation; (ii) interaction with the orphan receptor Nurr1 in development of mesencephalic dopaminergic neurons; (iii) modulation of nuclear FGFR1 interactions downstream of nerve growth factor (NGF) signaling. These coalitions demonstrate the versatility of non‐canonical, nuclear tyrosine kinase signaling in diverse cellular differentiation programs of neurons. This article is protected by copyright. All rights reserved
      PubDate: 2015-05-12T13:43:09.403151-05:
      DOI: 10.1002/jcp.25036
       
  • Control of the Normal and Pathological Development of Neural Stem and
           Progenitor Cells by the PC3/Tis21/Btg2 and Btg1 Genes
    • Authors: Laura Micheli; Manuela Ceccarelli, Stefano Farioli‐Vecchioli, Felice Tirone
      Abstract: The PC3/Tis21/Btg2 and Btg1 genes are transcriptional cofactors belonging to the Btg/Tob family, which regulate the development of several cell types, including neural precursors. We summarize here the actions of these genes on neural precursors in the adult neurogenic niches and the cognitive defects associated when their expression is altered. We consider also recent findings implicating them in neural and non‐neural tumors, since common developmental mechanisms are involved. PC3/Tis21 is required for the regulation of the maturation of stem and progenitor cells in the adult dentate gyrus and subventricular zone (SVZ), by controlling both their exit from the cell cycle and the ensuing terminal differentiation. Such actions are effected by regulating the expression of several genes, including cyclin D1, BMP4, Id3. In cerebellar precursors, however, PC3/Tis21 regulates chiefly their migration rather than proliferation or differentiation, with important implications for the onset of medulloblastoma, the cerebellar tumor. In fact PC3/Tis21 is a medulloblastoma‐suppressor, as its overexpression in cerebellar precursors inhibits this tumor; PC3/Tis21 shows anti‐tumor activity also in non‐neural tumors. Btg1 presents a different functional profile, as it controls proliferation in adult stem/progenitor cells of dentate gyrus and SVZ, where is required to maintain their self‐renewal and quiescence, but is apparently devoid of a direct control of their terminal differentiation or migration. Notably, physical exercise in Btg1‐null mice rescues the loss of proliferative capability occurring in older stem cells. Both genes could be further investigated as therapeutical targets, namely, Btg1 in the process of aging and PC3/Tis21 as a tumor‐suppressor. This article is protected by copyright. All rights reserved
      PubDate: 2015-05-12T13:42:51.81523-05:0
      DOI: 10.1002/jcp.25038
       
  • Revisiting Chaos Theorem to Understand the Nature of Mirnas in Response to
           Drugs of Abuse
    • Authors: Faten A. Taki; Xiaoping Pan, Baohong Zhang
      Abstract: Just like Matryoshka dolls, biological systems follow a hierarchical order that is based on dynamic bidirectional communication among its components. In addition to the convoluted inter‐relationships, the complexity of each component spans several folds. Therefore, it becomes rather challenging to investigate phenotypes resulting from these networks as it requires the integration of reductionistic and holistic approaches. One dynamic system is the transcriptome which comprises a variety of RNA species. Some, like microRNAs, have recently received a lot of attention. miRNAs are very pleiotropic and have been considered as therapeutic and diagnostic candidates in the biomedical fields. In this review, we survey miRNA profiles in response to drugs of abuse (DA) using 118 studies. After providing a summary of substance use disorders (Pandurangaiah et al.)‐related miRNAs, general patterns of miRNA signatures are compared among studies for each drug or for all drugs of abuse. Then, current challenges and drawbacks in the field are discussed. Finally, we provide support for considering miRNAs as a chaotic system in normal versus disrupted states particularly in SUD and propose an integrative approach for studying and analyzing miRNA data. This article is protected by copyright. All rights reserved
      PubDate: 2015-05-12T13:42:33.44171-05:0
      DOI: 10.1002/jcp.25037
       
  • GLP‐2 as Beneficial Factor in the Glucose Homeostasis in Mice Fed a
           High Fat Diet
    • Authors: Sara Baldassano; Francesca Rappa, Antonella Amato, Francesco Cappello, Flavia Mulè
      Abstract: Glucagon like peptide‐2 (GLP‐2) is a gastrointestinal hormone released in response to dietary nutrients, which acts through a specific receptor, the GLP‐2 receptor (GLP‐2R). The physiological effects of GLP‐2 are multiple, involving also the intestinal adaptation to high fat diet (HFD). In consideration of the well‐known relationship between chronic HFD and impaired glucose metabolism, in the present study we examined if the blocking of the GLP‐2 signaling by chronic treatment with the GLP‐2R antagonist, GLP‐2 (3–33), leads to functional consequences in the regulation of glucose metabolism in HFD‐fed mice. Compared with animals fed standard diet (STD), mice at the tenth week of HFD showed hyperglycaemia, glucose intolerance, high plasma insulin level after glucose load, increased pancreas weight and β cell expansion, but not insulin resistance. In HFD fed mice, GLP‐2 (3‐33) treatment for four weeks (from the sixth to the tenth week of diet) did not affect fasting glycaemia, but it significantly increased the glucose intolerance, both fasting and glucose‐induced insulin levels, and reduced the sensitivity to insulin leading to insulin‐resistance. In GLP‐2 (3‐33)‐treated HFD mice pancreas was significantly heavier and displayed a significant increase in β‐cell mass in comparison with vehicle‐treated HFD mice. In STD mice, the GLP‐2 (3‐33) treatment did not affect fasted or glucose‐stimulated glycemia, insulin, insulin sensitivity, pancreas weight and beta cell mass. The present study suggests that endogenous GLP‐2 may act as a protective factor against the dysregulation of the glucose metabolism that occurs in HFD mice, because GLP‐2 (3‐33) worsens glucose metabolism disorders. This article is protected by copyright. All rights reserved
      PubDate: 2015-05-12T13:42:23.527174-05:
      DOI: 10.1002/jcp.25039
       
  • Kinin B1 Receptor Deletion Affects Bone Healing in Type 1 Diabetic Mice
    • Authors: Natália P. Cignachi; João B. Pesquero, Rogério B. Oliveira, Adriana Etges, Maria M. Campos
      Abstract: The effects of kinin B1 receptor (B1R) deletion were examined on femur bone regeneration in streptozotocin (STZ)‐type 1 diabetes. Diabetes induction in wild‐type C57/BL6 (WTC57BL6) mice led to decrease in body weight and hyperglycemia, compared to the non‐diabetic group of the same strain. The lack of B1R did not affect STZ‐elicited body weight loss, but partially prevented hyperglycemia. Diabetic mice had a clear delay in bone regeneration, and displayed large areas of loose connective tissue within the defects, with a reduced expression of the mineralization‐related protein osteonectin, when compared to the non‐diabetic WTC57/BL6. The non‐diabetic and diabetic B1R knockout (B1RKO) mice had bone regeneration levels and osteonectin expression comparable to that seen in control WTC57/BL6 mice. WTC57/BL6 STZ‐diabetic mice also showed a marked reduction of collagen contents, with increased immunolabelling for the apoptosis marker caspase‐3, whereas diabetic B1RKO had collagen levels and caspase‐3 activity comparable to those observed in non‐diabetic WTC57/BL6 or B1RKO mice. No significant difference was detected in the number of tartrate‐resistant acid phosphatase (TRAP)‐stained cells, or in RANK/RANKL/OPG system immunolabelling throughout the experimental groups. Data bring novel evidence on the relevance of kinin B1R under type 1 diabetes with regards to its role in bone regeneration. This article is protected by copyright. All rights reserved
      PubDate: 2015-05-12T13:26:09.52783-05:0
      DOI: 10.1002/jcp.25034
       
  • Protein Kinase C Is Involved in the Induction of ATP‐Binding
           Cassette Transporter A1 Expression by Liver X Receptor/Retinoid X Receptor
           Agonist in Human Macrophages
    • Authors: Etimad A. Huwait; Nishi N. Singh, Daryn R. Michael, Thomas S. Davies, Joe W. E. Moss, Dipak P. Ramji
      Abstract: The transcription of the ATP‐binding cassette transporter A1 (ABCA1) gene, which plays a key anti‐atherogenic role, is known to be induced by agonists of liver X receptors (LXRs). LXRs form obligate heterodimers with retinoid X receptors (RXRs) and interact with their recognition sequences in the regulatory regions of key genes implicated in the control of cholesterol, fatty acid and glucose homeostasis. We have previously shown a novel role for c‐Jun N‐terminal kinase (JNK) and phosphoinositide 3‐kinase (PI3K) in the LXRs‐mediated induction of macrophage gene expression. Protein kinase C (PKC) is often found to regulate the action of nuclear receptors and cross talk between this kinase family and JNK and/or PI3K has been shown in several settings. We have therefore investigated a potential role for PKC in the action of LXR/RXR agonist 22‐(R)‐hydroxycholesterol (22‐(R)‐HC)/9‐cis‐retinoic acid (9cRA) in THP‐1 macrophages, including the induction of ABCA1 expression. The pan PKC inhibitor bisindoylmaleimide was found to attenuate the induction of ABCA1 protein expression, the activation of the JNK signaling pathway and the stimulation of activator protein‐1 (AP‐1) DNA binding activity in macrophages treated with 22‐(R)‐HC and 9cRA. The role of PKC in the action of these ligands was confirmed further by the use of more isotype‐specific inhibitors. These studies therefore reveal a potentially important role for PKC in the action of 22‐(R)‐HC and 9cRA in human macrophages. This article is protected by copyright. All rights reserved
      PubDate: 2015-05-07T18:01:38.140641-05:
      DOI: 10.1002/jcp.25157
       
  • S100A8, an Oocyte‐Specific Chemokine, Directs the Migration of
           Ovarian Somatic Cells During Mice Primordial Follicle Assembly
    • Authors: Zhen Teng; Chao Wang, Yijing Wang, Hua Zhang, Kun Huang, Xi Xiang, Wanbao Niu, Lizhao Feng, Lihua Zhao, Hao Yan, Guoliang Xia
      Abstract: In the mammalian ovaries, the primordial follicle pool determines the reproductive capability over the lifetime of a female. The primordial follicle is composed of two cell members, namely the oocyte and the pre‐granulosa cells that encircle the oocyte. However, it is unclear what factors are involved in the reorganization of the two distinct cells into one functional unit. This study was performed to address this issue. Firstly, in an in vitro reconstruction system, dispersed ovarian cells from murine fetal ovaries at 19.0 days post coitum (dpc) reassembled into follicle‐like structures, independent of the physical distance between the cells, implying that either oocytes or ovarian somatic cells (OSCs) were motile. We then carried out a series of transwell assay experiments, and determined that it was in fact 19.0 dpc OSCs (as opposed to oocytes), which exhibited a significant chemotactic response to both fetal bovine serum and oocytes themselves. We observed that S100A8, a multi‐functional chemokine, may participate in the process as it is mainly expressed in oocytes within the cysts/plasmodia. S100A8 significantly promoted the number of migrating OSCs by 2.5 times in vitro, of which 66.9% were FOXL2 protein‐positive cells, implying that the majority of motile OSCs were pre‐granulosa cells. In addition, an S100A8‐specific antibody inhibited the formation of follicle‐like reconstruction cell mass in vitro. And, the primordial follicle formation was reduced when S100a8‐specific siRNA was applied onto in vitro cultured 17.5 dpc ovary. Therefore, S100A8 could be a chemokine of oocyte origin, which attracts OSCs to form the primordial follicles. This article is protected by copyright. All rights reserved
      PubDate: 2015-05-06T17:18:24.367503-05:
      DOI: 10.1002/jcp.25032
       
  • Possible Involvement of Palmitate in Pathogenesis of Periodontitis
    • Authors: Yosuke Shikama; Yasusei Kudo, Naozumi Ishimaru, Makoto Funaki
      Abstract: Type 2 diabetes (T2D) is characterized by decreased insulin sensitivity and higher concentrations of free fatty acids (FFAs) in plasma. Among FFAs, saturated fatty acids (SFAs), such as palmitate, have been suggested to promote inflammatory responses. Although many epidemiological studies have shown a link between periodontitis and T2D, little is known about the clinical significance of SFAs in periodontitis. In this study, we showed that gingival fibroblasts have cell‐surface expression of CD36, which is also known as FAT/fatty acid translocase. Moreover, CD36 expression was increased in gingival fibroblasts of high‐fat diet‐induced T2D model mice, compared with gingival fibroblasts of mice fed a normal diet. DNA microarray analysis revealed that palmitate increased mRNA expression of pro‐inflammatory cytokines and chemokines in human gingival fibroblasts (HGF). Consistent with these results, we confirmed that palmitate induced interleukin (IL)‐6, IL‐8, and CXCL1 secretion in HGF, using a cytokine array and ELISA. SFAs, but not an unsaturated fatty acid, oleate, induced IL‐8 production. Docosahexaenoic acid (DHA), which is one of the omega‐3 polyunsaturated fatty acids, significantly suppressed palmitate‐induced IL‐6 and IL‐8 production. Treatment of HGF with a CD36 inhibitor also inhibited palmitate‐induced pro‐inflammatory responses. Finally, we demonstrated that Porphyromonas gingivalis (P.g.) lipopolysaccharide and heat‐killed P.g. augmented palmitate‐induced chemokine secretion in HGF. These results suggest a potential link between SFAs in plasma and the pathogenesis of periodontitis. This article is protected by copyright. All rights reserved
      PubDate: 2015-04-29T06:27:24.646609-05:
      DOI: 10.1002/jcp.25029
       
  • c‐Src‐dependent transactivation of EGFR mediates
           CORM‐2‐induced HO‐1 expression in human tracheal smooth
           muscle cells
    • Authors: Chuen‐Mao Yang; Chih‐Chung Lin, I‐Ta Lee, Chih‐Kai Hsu, Yu‐Chen Tai, Hsi‐Lung Hsieh, Pei‐Ling Chi, Andli‐Der Hsiao
      Abstract: Carbon monoxide (CO), a reaction product of the cytoprotective heme oxygenase (HO)‐1, displays an anti‐inflammatory effect in various cellular injuries, but the precise mechanisms of HO‐1 expression remain unknown. We used the transition metal carbonyl compound carbon monoxide‐releasing molecule‐2 (CORM‐2) that acts as carbon monoxide donor. The effects of CORM‐2 on expression of HO‐1 in human tracheal smooth muscle cells (HTSMCs) were determined by Western blot, real‐time PCR, and promoter activity assay. In HTSMCs, CORM‐2 activated Nrf2 through the activation of a c‐Src/EGFR/PI3K/Akt‐dependent pathway, resulting in HO‐1 expression. We showed that CORM‐2‐induced HO‐1 protein and mRNA levels were inhibited by the inhibitor of c‐Src (PP1 or SU6656), EGFR (AG1478), PI3K (LY294002), Akt (SH‐5), JNK1/2 (SP600125), or p38 MAPK (SB202190) and transfection with siRNA of c‐Src, EGFR, Akt, p38, JNK2, or Nrf2 in HTSMCs. We also showed that CORM‐2 stimulated c‐Src, EGFR, Akt, p38 MAPK, and JNK1/2 phosphorylation. CORM‐2 also enhanced Nrf2 translocation from the cytosol to the nucleus and antioxidant response element (ARE) promoter activity. Moreover, CORM‐2 mediated p38 MAPK and JNK1/2 activation via a c‐Src/EGFR/PI3K/Akt pathway, which further enhanced Nrf2 activation and translocation. Finally, we observed that CORM‐2 induced in vivo binding of Nrf2 to the HO‐1 promoter. CORM‐2 activates the c‐Src/EGFR/PI3K/Akt/JNK1/2 and p38 MAPK pathways, which in turn trigger Nrf2 activation and ultimately induces HO‐1 expression in HTSMCs. Thus, the HO‐1/CO system might be potential therapeutics in airway diseases. This article is protected by copyright. All rights reserved
      PubDate: 2015-04-29T06:27:07.927948-05:
      DOI: 10.1002/jcp.24912
       
  • The Immunomodulatory and Therapeutic Effects of Mesenchymal Stromal Cells
           for Acute Lung Injury and Sepsis
    • Authors: Mirabelle S.H. Ho; Shirley H.J. Mei, Duncan J. Stewart
      Abstract: It is increasingly recognized that immunomodulation represents an important mechanism underlying the benefits of many stem cell therapies, rather than the classical paradigm of transdifferentiation and cell replacement. In the former paradigm, the beneficial effects of cell therapy result from paracrine mechanism(s) and/or cell‐cell interaction as opposed to direct engraftment and repair of diseased tissue and/or dysfunctional organs. Depending on the cell type used, components of the secretome, including microRNA (miRNA) and extracellular vesicles, may be able to either activate or suppress the immune system even without direct immune cell contact. Mesenchymal stromal cells (MSCs), also referred to as mesenchymal stem cells, are found not only in the bone marrow, but in a wide variety of organs and tissues. In addition to any direct stem cell activities, MSCs were the first stem cells recognized to modulate immune response, and therefore they will be the focus of this review. Specifically, MSCs appear to be able to effectively attenuate acute and protracted inflammation via interactions with components of both innate and adaptive immune systems. To date, this capacity has been exploited in a large number of pre‐clinical studies and MSC immunomodulatory therapy has been attempted with various degrees of success in a relatively large number of clinical trials. Here we will explore the various mechanism employed by MSCs to effect immunosuppression as well as review the current status of its use to treat excessive inflammation in the context of acute lung injury and sepsis in both pre‐clinical and clinical settings. This article is protected by copyright. All rights reserved
      PubDate: 2015-04-25T06:46:25.357218-05:
      DOI: 10.1002/jcp.25028
       
  • Treg(s) in Cancer: Friends or Foe'
    • Authors: Dominik Wolf; Sieghart Sopper, Andreas Pircher, Guenther Gastl, Anna Maria Wolf
      Abstract: Immune escape is a hallmark of cancer. Regulatory T cells (Treg) have been described to maintain peripheral tolerance. The role of Treg in cancer is ambiguous, as they are central inhibitory regulators in solid tumors, whereas during inflammation‐driven tumorigenesis they prevent cancer initiation by restraining inflammation. As a consequence, under conditions with chronic inflammation that may initiate malignant transformation, application rather than depletion of Treg may be helpful. In solid tumors, however, the success story of immune‐activating antibodies targeting checkpoint molecules of T cell activation fuels the hope that Treg inactivation or depletion may additionally boost anti‐tumor immune response. In this review we summarize important aspects on the dual role of Treg in cancer to provide a rationale for future Treg targeting attempts. This article is protected by copyright. All rights reserved
      PubDate: 2015-04-25T06:46:07.296271-05:
      DOI: 10.1002/jcp.25016
       
  • Para‐Cresyl Sulfate Acutely Impairs Vascular Reactivity and Induces
           Vascular Remodeling
    • Authors: Priscilla Gross; Ziad A. Massy, Lucie Henaut, Cédric Boudot, Joanna Cagnard, Cécilia March, Saïd Kamel, Tilman B. Drueke, Isabelle Six
      Abstract: Chronic kidney disease (CKD) is characterized by vascular remodeling and the retention of uremic toxins, several of which are independently associated with the high cardiovascular mortality rate in CKD patients. Whether the association between these uremic toxins and cardiovascular mortality is due to induction of vascular dysfunction and resulting vascular remodeling remains to be determined. This study evaluates the effects of para‐cresyl sulfate (PCS), a newly identified uremic toxin, on vascular function and remodeling. PCS acutely induced oxidative stress in both endothelial and vascular smooth muscle cells, with a maximal effect at 0.15 mM, corresponding to the mean ‘uremic’ concentration found in dialysis patients. PCS significantly increased within 30 min phenylephrine‐induced contraction of mouse thoracic aorta, through direct activation of rho‐kinase, independently of oxidative stress induction, as demonstrated by the capacity of rho‐kinase inhibitor Y‐27632 to abolish this effect. After exposure of the aorta to PCS for 48 h, we observed inward eutrophic remodeling, a hallmark of uremic vasculopathy characterized by a reduction of the area of both lumen and media, with unchanged media/lumen ratio. In conclusion, elevated PCS concentrations such as those observed in CKD patients, by promoting both vascular dysfunction and vascular remodeling, may contribute to the development of hypertension and to cardiovascular mortality in CKD. This article is protected by copyright. All rights reserved
      PubDate: 2015-04-22T00:19:35.852625-05:
      DOI: 10.1002/jcp.25018
       
  • Transgenic Expression of Osteoactivin/gpnmb Enhances Bone Formation in
           Vivo and Osteoprogenitor Differentiation ex Vivo
    • Authors: Nagat Frara; Samir M. Abdelmagid, Gregory R. Sondag, Fouad M. Moussa, Vanessa R. Yingling, Thomas A. Owen, Steven N. Popoff, Mary F. Barbe, Fayez F. Safadi
      Abstract: Initial identification of osteoactivin (OA)/glycoprotein non‐melanoma clone B (gpnmb) was demonstrated in an osteopetrotic rat model, where OA expression was increased 3‐fold in mutant bones, compared to normal. OA mRNA and protein expression increase during active bone regeneration post‐fracture, and primary rat osteoblasts show increased OA expression during differentiation in vitro. To further examine OA/gpnmb as an osteoinductive agent, we characterized the skeletal phenotype of transgenic mouse overexpressing OA/gpnmb under the CMV‐promoter (OA‐Tg). Western blot analysis showed increased OA/gpnmb in OA‐Tg osteoblasts, compared to wild‐type (WT). In OA‐Tg mouse femurs versus WT littermates, micro‐CT analysis showed increased trabecular bone volume and thickness, and cortical bone thickness; histomorphometry showed increased osteoblast numbers, bone formation and mineral apposition rates in OA‐Tg mice; and biomechanical testing showed higher peak moment and stiffness. Given that OA/gpnmb is also over‐expressed in osteoclasts in OA‐Tg mice, we evaluated bone resorption by ELISA and histomorphometry, and observed decreased serum CTX‐1 and RANK‐L, and decreased osteoclast numbers in OA‐Tg, compared to WT mice, indicating decreased bone remodeling in OA‐Tg mice. The proliferation rate of OA‐Tg osteoblasts in vitro was higher, compared to WT, as was alkaline phosphatase staining and activity, the latter indicating enhanced differentiation of OA‐Tg osteoprogenitors. Quantitative RT‐PCR analysis showed increased TGF‐β1 and TGF‐β receptors I and II expression in OA‐Tg osteoblasts, compared to WT. Together, these data suggest that OA overexpression has an osteoinductive effect on bone mass in vivo and stimulates osteoprogenitor differentiation ex vivo. This article is protected by copyright. All rights reserved
      PubDate: 2015-04-22T00:19:18.454348-05:
      DOI: 10.1002/jcp.25020
       
  • Myeloid Zinc Finger 1 (MZF‐1) Regulates Expression of the CCN2/CTGF
           and CCN3/NOV Genes in the Hematopoietic Compartment
    • Authors: Richard T. Piszczatowski; Brian J. Rafferty, Andre Rozado, James V. Parziale, Nathan H. Lents
      Abstract: Connective Tissue Growth Factor (CCN2/CTGF) and Nephroblastoma Overexpressed (CCN3/NOV) execute key functions within the hematopoietic compartment. Both are abundant in the bone marrow stroma, which is a niche for hematopoiesis and supports marrow function. Roles for 1,25‐dihydroxyvitamin D3 (calcitriol) and all‐trans retinoic acid in the bone marrow have also been elucidated. Interestingly, some of the annotated roles of these vitamins overlap with established functions of CCN2 and CCN3. Yet, no factor has been identified that unifies these observations. In this study, we report the regulation of the CTGF and NOV genes by Myeloid Zinc Finger‐1 (MZF‐1), a hematopoietic transcription factor. We show the interaction of MZF‐1 with the CTGF and NOV promoters in several cell types. Up‐regulation of MZF‐1 via calcitriol and vitamin A induces expression of CTGF and NOV, implicating a role for these vitamins in the functions of these two genes. Lastly, knockdown of MZF1 reduces levels of CTGF and NOV. Collectively, our results argue that MZF‐1 regulates the CTGF and NOV genes in the hematopoietic compartment, and may be involved in their respective functions in the stroma. This article is protected by copyright. All rights reserved
      PubDate: 2015-04-22T00:18:59.642223-05:
      DOI: 10.1002/jcp.25021
       
  • Functional Characterization of Nupr1L, a novelp53‐Regulated Isoform
           of the High Mobility Group (HMG)‐Related Protumoral Protein Nupr1
    • Authors: Maria Belen Lopez; Maria Noé Garcia, Daniel Grasso, Jennifer Bintz, Maria Inés Molejon, Gabriel Velez, Gwen Lomberk, Jose Luis Neira, Raul Urrutia, Juan Iovanna
      Abstract: We have previously demonstrated a crucial role of NUPR1 in tumor development and progression. In this work we report the functional characterization of a novel Nupr1‐like isoform (NUPR1L) and its functional interaction with the protumoral factor NUPR1. Through the use of primary sequence analysis, threading, and homology‐based molecular modeling as well as expression and immunolocalization, studies reveal that NUPR1L displays properties which are similar to member of the HMG‐like family of chromatin regulators including its ability to translocate to the cell nucleus and bind to DNA. Analysis of the NUPR1L promoter showed the presence of two p53‐response elements at positions ‐37 and ‐7 respectively. Experiments using reporter assays combined with site‐directed mutagenesis and using cells with controllable p53 expression demonstrate that both of these sequences are responsible for the regulation of NUPR1L expression by p53. Congruently, NUPR1L gene expression is activated in response to DNA damage induced by Oxaliplatin treatment or cell cycle arrest induced by serum starvation, two well‐validated methods to achieve p53 activation. Interestingly, expression of NUPR1L down‐regulates the expression of NUPR1, its closely related protumoral isoform, by a mechanism that involves the inhibition of its promoter activity. At the cellular level, overexpression of NUPR1L induces G1 cell cycle arrest and a decrease in their cell viability, an effect that is mediated, at least in part, by down‐regulating NUPR1 expression. Combined these experiments constitute the first functional characterization of NUPR1L as a new p53‐induced gene which negatively regulates the protumoral factor NUPR1. This article is protected by copyright. All rights reserved
      PubDate: 2015-04-22T00:18:41.462726-05:
      DOI: 10.1002/jcp.25022
       
  • Melanoma Treatments: Advances and Mechanisms
    • Authors: Alexander Marzuka; Laura Huang, Nicholas Theodosakis, Marcus Bosenberg
      Abstract: Advances in the understanding of the molecular pathogenesis of melanoma and in cancer immunology have led to the rational design and recent clinical implementation of a variety of novel therapies for metastatic melanoma. BRAF and MEK inhibitors that target the MAPK pathway have high rates of clinical response in BRAF‐mutant melanoma. Therapies that modulate the immune response to melanoma, including monoclonal antibodies that interfere with pathways that inhibit T‐cell function, have been shown to be effective in melanoma. Lessons learned from these encouraging results are driving the development of new treatments for melanoma and other cancers. This review will focus on the science and clinical findings related to targeted therapies that inhibit BRAF or MEK as well as the immunotherapies that block the CTLA‐4 or PD‐1 pathways. Other experimental and combinatorial therapeutic approaches will also be discussed. This article is protected by copyright. All rights reserved
      PubDate: 2015-04-22T00:18:24.092711-05:
      DOI: 10.1002/jcp.25019
       
  • TRPC1 Regulates Calcium‐Activated Chloride Channels in Salivary
           Gland Cells
    • Authors: Yuyang Sun; Lutz Birnbaumer, Brij B Singh
      Abstract: Calcium‐activated chloride channel (CaCC) plays an important role in modulating epithelial secretion. It has been suggested that in salivary tissues, sustained fluid secretion is dependent on Ca2+ influx that activates ion channels such as CaCC to initiate Cl‐ efflux. However direct evidence as well as the molecular identity of the Ca2+ channel responsible for activating CaCC in salivary tissues is not yet identified. Here we provide evidence that in human salivary cells, an outward rectifying Cl‐ current was activated by increasing [Ca2+]i, which was inhibited by the addition of pharmacological agents niflumic acid (NFA), an antagonist of CaCC, or T16Ainh‐A01, a specific TMEM16a inhibitor. Addition of thapsigargin (Tg), that induces store‐depletion and activates TRPC1‐mediated Ca2+ entry, potentiated the Cl‐ current, which was inhibited by the addition of a non‐specific TRPC channel blocker SKF96365 or removal of external Ca2+. Stimulation with Tg also increased plasma membrane expression of TMEM16a protein, which was also dependent on Ca2+ entry. Importantly, in salivary cells, TRPC1 silencing, but not that of TRPC3, inhibited CaCC especially upon store depletion. Moreover, primary acinar cells isolated from submandibular gland also showed outward rectifying Cl‐ currents upon increasing [Ca2+]i. These Cl‐ currents were again potentiated with the addition of Tg, but inhibited in the presence of T16Ainh‐A01. Finally, acinar cells isolated from the submandibular glands of TRPC1 knockout mice showed significant inhibition of the outward Cl‐ currents without decreasing TMEM16a expression. Together the data suggests that Ca2+ entry via the TRPC1 channels is essential for the activation of CaCC. This article is protected by copyright. All rights reserved
      PubDate: 2015-04-21T06:44:39.122861-05:
      DOI: 10.1002/jcp.25017
       
  • A New Medical Device Rigeneracons Allows to Obtain Viable
           Micro‐Grafts From Mechanical Disaggregation of Human Tissues
    • Authors: Letizia Trovato; Manuela Monti, Claudia del Fante, Marila Cervio, Milla Lampinen, Lucia Ambrosio, Carlo Alberto Redi, Cesare Perotti, Esko Kankuri, Gennaro Ambrosio, Ruggero Rodriguez Y. Baena, Giuseppe Pirozzi, Antonio Graziano
      Pages: 2299 - 2303
      Abstract: Autologous graft is considered the gold standard of graft materials; however, this approach is still limited due to both small amount of tissue that can be collected and to reduced cell viability of cells that can be obtained. The aim of this preliminary study was to demonstrate the efficacy of an innovative medical device called Rigeneracons® (CE certified Class I) to provide autologous micro‐grafts immediately available to be used in the clinical practice. Moreover, Rigeneracons® is an instrument able to create micro‐grafts enriched of progenitors cells which maintain their regenerative and differentiation potential. We reported preliminary data about viability cell of samples derived from different kind of human tissues, such as periosteum, cardiac atrial appendage biopsy, and lateral rectus muscle of eyeball and disaggregated by Rigeneracons®. In all cases we observed that micro‐grafts obtained by Rigeneracons® displayed high cell viability. Furthermore, by cell characterization of periosteum samples, we also evidenced an high positivity to mesenchymal cell markers, suggesting an optimal regenerative potential. J. Cell. Physiol. 230: 2299–2303, 2015. © 2015 Wiley Periodicals, Inc.
      PubDate: 2015-06-23T17:18:35.422372-05:
      DOI: 10.1002/jcp.24973
       
  • Genetic Architecture of Sexual Dimorphism in Humans
    • Authors: Nichole Rigby; Rob J. Kulathinal
      Pages: 2304 - 2310
      Abstract: Males and females differ across a broad spectrum of morphological, physiological, and behavioral characters. In fact, sexually dimorphic traits typically contribute the largest component of phenotypic variance in most taxa that use sex to reproduce. However, we know very little about the mechanisms that maintain these dimorphic states and how these sexually dimorphic traits evolve. Here, we review our current knowledge of the underlying genetic basis of sexual dimorphism in humans. First, we briefly review the etiology of sex differences starting from sex determination's initial switch early in embryogenesis. We then survey recent sex‐biased transcriptomic expression literature in order to provide additional insight into the landscape of sex‐biased gene expression in both gonadal and non‐gonadal tissues: from overall prevalence to tissue specificity to conservation across species. Finally, we discuss implications of sex‐biased genetic architecture to human health and disease in light of the National Institute of Health's recently proposed initiative to promote study samples from both sexes. J. Cell. Physiol. 230: 2304–2310, 2015. © 2015 Wiley Periodicals, Inc.
      PubDate: 2015-06-23T17:18:50.187281-05:
      DOI: 10.1002/jcp.24979
       
  • Can Brown Fat Win the Battle Against White Fat?
    • Authors: Sawsan Elattar; Ande Satyanarayana
      Pages: 2311 - 2317
      Abstract: A rapid growth in the overweight and obese population in the last few decades suggest that the current diet, exercise, awareness or drug strategies are still not effectively restraining the obesity epidemic. Obesity results from increased energy intake, and the body's energy balance shifts towards energy abundance. Therefore, current research is focused on developing new strategies aimed at increasing energy expenditure. As a result, brown adipose tissue (BAT) is receiving tremendous attention since the major function of BAT is to dissipate energy as heat. For example, mouse models that have increased BAT activity or increased numbers of brown‐like adipocytes within the white adipose tissue (WAT) are lean and protected from obesity. Alternatively, mouse models that lack BAT activity are more susceptible to age and diet‐induced obesity. However, a significant loss of BAT mass during the natural growth process in humans has created enormous challenges in effectively utilizing this tissue to increase energy expenditure. New strategies are primarily focused on expanding the BAT mass and/or activating the existing BAT. In this regard, recent finding that expression of early B cell factor‐2 (Ebf2) reprograms the white pre‐adipocytes into brown adipocytes is a significant break‐through in developing BAT‐mediated strategies to treat obesity. Here we review the major biological functions of WAT and BAT, which play critical but opposing roles in the energy spectrum, energy storage versus energy expenditure, and we evaluate whether activation and/or expansion of BAT is practically achievable to treat obesity in humans. J. Cell. Physiol. 230: 2311–2317, 2015. © 2015 Wiley Periodicals, Inc.
      PubDate: 2015-06-23T17:18:41.768469-05:
      DOI: 10.1002/jcp.24986
       
  • Regulation of MDM2 Stability After DNA Damage
    • Authors: Jiaqi Li; Manabu Kurokawa
      Pages: 2318 - 2327
      Abstract: Cells in our body are constantly exposed to various stresses and threats to their genomic integrity. The tumor suppressor protein p53 plays a critical role in successful defense against these threats by inducing apoptotic cell death or cell cycle arrest. In unstressed conditions, p53 levels and activity must be kept low to prevent lethal activation of apoptotic and senescence pathways. However, upon DNA damage or other stressors, p53 is released from its inhibitory state to induce an array of apoptosis and cell cycle genes. Conversely, inactivation of p53 could promote unrestrained tumor proliferation and failure to appropriately undergo apoptotic cell death, which could, in turn, lead to carcinogenesis. The ubiquitin E3 ligase MDM2 is the most critical inhibitor of p53 that determines the cellular response to various p53‐activating agents, including DNA damage. MDM2 activity is controlled by post‐translational modifications, especially phosphorylation. However, accumulating evidence suggests that MDM2 is also regulated at the level of protein stability, which is controlled by the ubiquitin‐proteasome pathway. Here, we discuss how MDM2 can be regulated in response to DNA damage with particular focus on the regulation of MDM2 protein stability. J. Cell. Physiol. 230: 2318–2327, 2015. © 2015 Wiley Periodicals, Inc.
      PubDate: 2015-06-23T17:18:54.556412-05:
      DOI: 10.1002/jcp.24994
       
  • Antidepressant Effects of Exercise: A Role for the
           Adiponectin‐PGC‐1α‐kynurenine Triad?
    • Authors: Fabian Sanchis‐Gomar; Carme Pérez Quilis, Alejandro Lucia
      Pages: 2328 - 2329
      PubDate: 2015-06-23T17:18:53.975483-05:
      DOI: 10.1002/jcp.24964
       
  • Transcription Factor Tfe3 Directly Regulates Pgc‐1alpha in Muscle
    • Authors: Nunciada Salma; Jun S. Song, Zoltan Arany, David E. Fisher
      Pages: 2330 - 2336
      Abstract: The microphthalmia (MiT) family of transcription factors is an important mediator of metabolism. Family members Mitf and Tfeb directly regulate the expression of the master regulator of metabolism, peroxisome‐proliferator activated receptor gamma coactivator‐1 alpha (Pgc‐1alpha), in melanomas and in the liver, respectively. Pgc‐1alpha is enriched in tissues with high oxidative capacity and plays an important role in the regulation of mitochondrial biogenesis and cellular metabolism. In skeletal muscle, Pgc‐1alpha affects many aspects of muscle functionally such as endurance, fiber‐type switching, and insulin sensitivity. Tfe3 also regulates muscle metabolic genes that enhance insulin sensitivity in skeletal muscle. Tfe3 has not yet been shown to regulate Pgc‐1alpha expression. Our results reported here show that Tfe3 directly regulates Pgc‐1alpha expression in myotubes. Tfe3 ectopic expression induces Pgc‐1alpha, and Tfe3 silencing suppresses Pgc‐1alpha expression. This regulation is direct, as shown by Tfe3's binding to E‐boxes on the Pgc‐1alpha proximal promoter. We conclude that Tfe3 is a critical transcription factor that regulates Pgc‐1alpha gene expression in myotubes. Since Pgc‐1alpha coactivates numerous biological programs in diverse tissues, the regulation of its expression by upstream transcription factors such Tfe3 implies potential opportunities for the treatment of diseases where modulation of Pgc‐1alpha expression may have important clinical outcomes. J. Cell. Physiol. 230: 2330–2336, 2015. © 2015 Wiley Periodicals, Inc.
      PubDate: 2015-06-23T17:18:49.416616-05:
      DOI: 10.1002/jcp.24978
       
  • Roles for Histone Acetylation in Regulation of Telomere Elongation and
           Two‐cell State in Mouse ES Cells
    • Authors: Jiameng Dan; Jiao Yang, Yifei Liu, Andrew Xiao, Lin Liu
      Pages: 2337 - 2344
      Abstract: Mammalian telomeres and subtelomeres are marked by heterochromatic epigenetic modifications, including repressive DNA methylation and histone methylation (e.g., H3K9me3 and H4K20me3). Loss of these epigenetic marks results in increased rates of telomere recombination and elongation. Other than these repressive epigenetic marks, telomeric and subtelomeric H3 and H4 are underacetylated. Yet, whether histone acetylation also regulates telomere length has not been directly addressed. We thought to test the effects of histone acetylation levels on telomere length using histone deacetylase (HDAC) inhibitor (sodium butyrate, NaB) that mediates histone hyperacetylation and histone acetyltransferase (HAT) inhibitor (C646) that mediates histone hypoacetylation. We show that histone hyperacetylation dramatically elongates telomeres in wild‐type ES cells, and only slightly elongates telomeres in Terc−/− ES cells, suggesting that Terc is involved in histone acetylation‐induced telomere elongation. In contrast, histone hypoacetylation shortens telomeres in both wild‐type and Terc−/− ES cells. Additionally, histone hyperacetylation activates 2‐cell (2C) specific genes including Zscan4, which is involved in telomere recombination and elongation, whereas histone hypoacetylation represses Zscan4 and 2C genes. These data suggest that histone acetylation levels affect the heterochromatic state at telomeres and subtelomeres, and regulate gene expression at subtelomeres, linking histone acetylation to telomere length maintenance. J. Cell. Physiol. 230: 2337–2344, 2015. © 2015 Wiley Periodicals, Inc.
      PubDate: 2015-06-23T17:18:42.444873-05:
      DOI: 10.1002/jcp.24980
       
  • TNFα Post‐Translationally Targets ZnT2 to Accumulate Zinc in
           Lysosomes
    • Authors: Stephen R. Hennigar; Shannon L. Kelleher
      Pages: 2345 - 2350
      Abstract: Mammary epithelial cells undergo widespread lysosomal‐mediated cell death (LCD) during early mammary gland involution. Recently, we demonstrated that tumor necrosis factor‐α (TNFα), a cytokine released during early involution, redistributes the zinc (Zn) transporter ZnT2 to accumulate Zn in lysosomes and activate LCD and involution. The objective of this study is to determine how TNFα retargets ZnT2 to lysosomes. We tested the hypothesis that TNFα signaling dephosphorylates ZnT2 to uncover a highly conserved dileucine motif (L294L) in the C‐terminus of ZnT2, allowing adaptor protein complex‐3 (AP‐3) to bind and traffic ZnT2 to lysosomes. Confocal micrographs showed that TNFα redistributed wild‐type (WT) ZnT2 from late endosomes (Pearson's coefficient = 0.202 ± 0.05 and 0.097 ± 0.03; P 
      PubDate: 2015-06-23T17:18:33.245511-05:
      DOI: 10.1002/jcp.24992
       
  • EGF Suppresses the Initiation and Drives the Reversion of
           TGF‐β1‐induced Transition in Hepatic Oval Cells Showing
           the Plasticity of Progenitor Cells
    • Authors: Ping Wang; AI‐Ting Yang, Min Cong, Tian‐Hui Liu, Dong Zhang, Jian Huang, Xiao‐Fei Tong, Sheng‐Tao Zhu, Yong Xu, Shu‐Zhen Tang, Bao‐En Wang, Hong Ma, Ji‐Dong Jia, Hong You
      Pages: 2362 - 2370
      Abstract: Transforming growth factor‐β1 (TGF‐β1) induces hepatic progenitors to tumor initiating cells through epithelial‐mesenchymal transition (EMT), thus raising an important drawback for stem cell–based therapy. How to block and reverse TGF‐β1‐induced transition is crucial for progenitors’ clinical application and carcinogenic prevention. Rat adult hepatic progenitors, hepatic oval cells, experienced E‐cadherin to N‐cadherin switch and changed to α‐smooth muscle actin (α‐SMA) positive cells after TGF‐β1 incubation, indicating EMT. When TGF‐β1 plus EGF were co‐administrated to these cells, EGF dose‐dependently suppressed the cadherin switch and α‐SMA expression. Interestingly, if EGF was applied to TGF‐β1‐pretreated cells, the cells that have experienced EMT could return to their epithelial phenotype. Abruption of EGF receptor revealed that EGF exerted its blockage and reversal effects through phosphorylation of ERK1/2 and Akt. These findings suggest an important attribute of EGF on opposing and reversing TGF‐β1 effects, indicating the plasticity of hepatic progenitors. J. Cell. Physiol. 230: 2362–2370, 2015. © 2015 Wiley Periodicals, Inc.
      PubDate: 2015-06-23T17:18:41.250239-05:
      DOI: 10.1002/jcp.24962
       
  • β‐Arrestin2 Contributes to Cell Viability and Proliferation via
           the Down‐Regulation of FOXO1 in Castration‐Resistant Prostate
           Cancer
    • Authors: Xiaolu Duan; Zhenzhen Kong, Yang Liu, Zhiwen Zeng, Shujue Li, Wenqi Wu, Weidong Ji, Bicheng Yang, Zhijian Zhao, Guohua Zeng
      Pages: 2371 - 2381
      Abstract: β‐Arrestin2 has been identified to act as a corepressor of androgen receptor (AR) signaling by binding to AR and serving as a scaffold to affect the activity and expression of AR in androgen‐dependent prostate cancer cells; however, little is known regarding its role in castration‐resistant prostate cancer (CRPC) progression. Here, our data demonstrated that β‐arrestin2 contributes to the cell viability and proliferation of CRPC via the downregulation of FOXO1 activity and expression. Mechanistically, in addition to its requirement for FOXO1 phosphorylation induced by IGF‐1, β‐arrestin2 could inhibit FOXO1 activity in an Akt‐independent manner and delay FOXO1 dephosphorylation through the inhibition of PP2A phosphatase activity and the attenuation of the interaction between FOXO1 and PP2A. Furthermore, β‐arrestin2 could downregulate FOXO1 expression via ubiquitylation and proteasomal degradation. Together, our results identified a novel role for β‐arrestin2 in the modulation of the CRPC progress through FOXO1. Thus, the characterization of β‐arrestin2 may represent an alternative therapeutic target for CRPC treatment. J. Cell. Physiol. 230: 2371–2381, 2015. © 2015 Wiley Periodicals, Inc.
      PubDate: 2015-06-23T17:18:52.984607-05:
      DOI: 10.1002/jcp.24963
       
  • Autophagy Mediates HBx‐Induced Nuclear Factor‐κB
           Activation and Release of IL‐6, IL‐8, and CXCL2 in Hepatocytes
           
    • Authors: Millore X.M. Luo; Sunny H. Wong, Matthew T.V. Chan, Le Yu, Sidney S.B. Yu, Feng Wu, Zhangang Xiao, Xiaojuan Wang, Lin Zhang, Alfred S.L. Cheng, Simon S.M. Ng, Francis K.L. Chan, Chi H. Cho, Jun Yu, Joseph J.Y. Sung, William K.K. Wu
      Pages: 2382 - 2389
      Abstract: Hepatitis B virus (HBV) and one of its encoded proteins, HBV X protein (HBx), have been shown to induce autophagy in hepatoma cells. Substantial evidence indicates that autophagy is a potent suppressor of inflammation. However, sporadic reports suggest that autophagy could promote pro‐inflammatory cytokine expression and inflammation in some biological contexts. Here, we show that overexpression of HBx induces LC3B‐positive autophagosome formation, increases autophagic flux and enhances the expression of ATG5, ATG7, and LC3B‐II in normal hepatocytes. Abrogation of autophagy by small interfering RNA against ATG5 and ATG7 prevents HBx‐induced formation of autophagosomes. Autophagy inhibition also abrogates HBx‐induced activation of nuclear factor‐κB (NF‐κB) and production of interleukin‐6 (IL‐6), IL‐8, and CXCL2. These findings suggest that autophagy is required for HBx‐induced NF‐κB activation and pro‐inflammatory cytokine production and could shed new light on the complex role of autophagy in the modulation of inflammation. J. Cell. Physiol. 230: 2382–2389, 2015. © 2015 Wiley Periodicals, Inc.
      PubDate: 2015-06-23T17:18:55.210053-05:
      DOI: 10.1002/jcp.24967
       
  • Silencing Angiopoietin‐Like Protein 4 (ANGPTL4) Protects Against
           Lipopolysaccharide‐Induced Acute Lung Injury Via Regulating SIRT1
           /NF‐kB Pathway
    • Authors: Liang Guo; Shaoying Li, Yunfeng Zhao, Pin Qian, Fuyun Ji, Lanlan Qian, Xueling Wu, Guisheng Qian
      Pages: 2390 - 2402
      Abstract: Lung inflammation and alveolar epithelial cell death are critical events in the development and progression of acute lung injury (ALI). Although angiopoietin‐like protein 4 (ANGPTL4) participates in inflammation, whether it plays important roles in ALI and alveolar epithelial cell inflammatory injury remains unclear. We therefore investigated the role of angptl4 in lipopolysaccharide (LPS)‐induced ALI and the associated mechanisms. Lentivirus‐mediated short interfering RNA targeted to the mouse angptl4 gene (AngsiRNA) and a negative control lentivirus (NCsiRNA) were intranasally administered to mice. Lung inflammatory injury and the underlying mechanisms for regulation of angptl4 on the LPS‐induced ALI were subsequently determined. We reported that angptl4 levels were increased both in human alveolar epithelial A549 cells and lung tissues obtained from a mouse model of LPS‐induced ALI. Angptl4 expression was induced by LPS in alveolar epithelial cells, whereas LPS‐induced lung inflammation (neutrophils infiltration in the lung tissues, tumor necrosis factor α, interleukin 6), lung permeability (lung wet/dry weight ratio and bronchoalveolar lavage fluid (BALF) protein concentration), tissue damage (caspase3 activation), and mortality rates were attenuated in AngsiRNA‐treated mice. The inflammatory reaction (tumor necrosis factor α, interleukin 6) and apoptosis rates were reduced in AngsiRNA(h)‐treated A549 cells. Moreover, angptl4 promoted NF‐kBp65 expression and suppressed SIRT1 expression both in mouse lungs and A549 cells. Additionally, SIRT1 antagonist nicotinamide (NAM) attenuated the inhibitory effects of AngsiRNA both on LPS‐induced NF‐kBp65 expression and IL6 expression. These findings suggest that silencing angptl4 protects against LPS‐induced ALI via regulating SIRT1/NF‐kB signaling pathway. J. Cell. Physiol. 230: 2390–2402, 2015. © 2015 Wiley Periodicals, Inc.
      PubDate: 2015-06-23T17:18:47.582575-05:
      DOI: 10.1002/jcp.24969
       
  • Transplanted Adipose‐Derived Stem Cells Ameliorate Testicular
           Dysfunction In A D‐Galactose–Induced Aging Rat Model
    • Authors: Chun Yang; Yi‐kuan Du, Jun Wang, Ping Luan, Qin‐lao Yang, Wen‐hua Huang, Lin Yuan
      Pages: 2403 - 2414
      Abstract: Glycation product accumulation during aging of slowly renewing tissues may be an important mechanism underlying aging of the testis. Adipose‐derived stem cells (ADSCs) have shown promise in a novel tissue regenerative technique and may have utility in treating sexual dysfunction. ADSCs have also been found to be effective in antiaging therapy, although the mechanism underlying their effects remains unknown. This study was designed to investigate the anti‐aging effect of ADSCs in a D‐galactose (D‐gal)–induced aging animal model and to clarify the underlying mechanism. Randomly selected 6‐week‐old male Sprague–Dawley rats were subcutaneously injected with D‐gal daily for 8 weeks. Two weeks after completion of treatment, D‐gal–induced aging rats were randomized to receive caudal vein injections of 3 × 106 5‐bromo 2′deoxy­uridine–labeled ADSCs or an equal volume of phosphate‐buffered saline. Serum testosterone level, steroidogenic enzymes (3‐β‐hydroxysteroid dehydrogenase), and superoxide dismutase (SOD) activity decreased significantly in aging rats compared with the control group; serum lipid peroxidation, spermatogenic cell apoptosis, and methane dicarboxylic aldehyde (MDA) expression increased significantly. ADSCs increased the SOD level and reduced the MDA level in the aging animal model and restored levels of serum testosterone, steroidogenic enzymes, and spermatogenic cell apoptosis. These results demonstrate that ADSCs can contribute to testicular regeneration during aging. ADSCs also provide functional benefits through glycation suppression and antioxidant effects in a rat model of aging. Although some ADSCs differentiated into Leydig cells, the paracrine pathway seems to play a main role in this process, resulting in the reduction of apoptosis. J. Cell. Physiol. 230: 2403–2414, 2015. © 2015 Wiley Periodicals, Inc.
      PubDate: 2015-06-23T17:18:36.481445-05:
      DOI: 10.1002/jcp.24970
       
  • Pantethine Alters Lipid Composition and Cholesterol Content of Membrane
           Rafts, With Down‐Regulation of CXCL12‐Induced T Cell Migration
           
    • Authors: Manuel van Gijsel‐Bonnello; Niyazi Acar, Yves Molino, Lionel Bretillon, Michel Khrestchatisky, Max de Reggi, Bouchra Gharib
      Pages: 2415 - 2425
      Abstract: Pantethine, a natural low‐molecular‐weight thiol, shows a broad activity in a large range of essential cellular pathways. It has been long known as a hypolipidemic and hypocholesterolemic agent. We have recently shown that it exerts a neuroprotective action in mouse models of cerebral malaria and Parkinson's disease through multiple mechanisms. In the present study, we looked at its effects on membrane lipid rafts that serve as platforms for molecules engaged in cell activity, therefore providing a target against inappropriate cell response leading to a chronic inflammation. We found that pantethine‐treated cells showed a significant change in raft fatty acid composition and cholesterol content, with ultimate downregulation of cell adhesion, CXCL12‐driven chemotaxis, and transendothelial migration of various T cell types, including human Jurkat cell line and circulating effector T cells. The mechanisms involved include the alteration of the following: (i) CXCL12 binding to its target cells; (ii) membrane dynamics of CXCR4 and CXCR7, the two CXCL12 receptors; and (iii) cell redox status, a crucial determinant in the regulation of the chemokine system. In addition, we considered the linker for activation of T cells molecule to show that pantethine effects were associated with the displacement from the rafts of the acylated signaling molecules which had their palmitoylation level reduced.. In conclusion, the results presented here, together with previously published findings, indicate that due to its pleiotropic action, pantethine can downregulate the multifaceted process leading to pathogenic T cell activation and migration. J. Cell. Physiol. 230: 2415–2425, 2015. © 2015 Wiley Periodicals, Inc.
      PubDate: 2015-06-23T17:18:44.311686-05:
      DOI: 10.1002/jcp.24971
       
  • Phosphorylation of Runx2, induced by cyclic mechanical tension via ERK1/2
           pathway, contributes to osteodifferentiation of human periodontal ligament
           fibroblasts
    • Authors: Dapeng Ren; Fulan Wei, Lihua Hu, Shuangyan Yang, Chunling Wang, Xiao Yuan
      Pages: 2426 - 2436
      Abstract: Occlusal force is an important stimulus for maintaining periodontal homeostasis. This is attributed to the quality of human periodontal ligament fibroblasts (hPDLFs) that could transfer occlusal force into biological signals modulating osteoblst differentiation. However, few studies investigated the mechanism of occlusal force‐induced osteodifferentiation of hPDLFs. In our study, we used the cyclic mechanical tension (CMT) at 10% elongation with 0.5 Hz to mimic occlusal force, and explored its effects on osteogenesis of hPDLFs. Firstly, elevated expressions of several osteoblast marker genes (Runx2, ATF4, SP7, OCN, and BSP), as well as activated ERK1/2 pathway were detected during CMT loading for 1, 3, 6, 12, 18, and 24 h. To gain further insight into how CMT contributed to those effects, we focused on the classic ERK1/2‐Runx2 pathway by inhibiting ERK1/2 and overexpressing Runx2. Our results reflected that Runx2 overexpression alone could induce osteodifferentiation of hPDLFs. Meanwhile, CMT loading could intensify while combined ERK1/2 blockage could weaken this process. Furthermore, we found that CMT promoted Runx2 transcription and phosphorylation via ERK1/2; protein level of phospho‐Runx2 (p‐Runx2), rather than Runx2, was in parallel with mRNA expressions of SP7, OCN, and BSP. Taken together, our study proved that p‐Runx2, elevated by CMT via ERK1/2 pathway, is the predominate factor in promoting osteoblast differentiation of hPDLFs. J. Cell. Physiol. 230: 2426–2436, 2015. © 2015 Wiley Periodicals, Inc.
      PubDate: 2015-06-23T17:18:34.738095-05:
      DOI: 10.1002/jcp.24972
       
  • In vivo sodium tungstate treatment prevents E‐cadherin loss induced
           by diabetic serum in HK‐2 cell line
    • Authors: Romina Bertinat; Pamela Silva, Elizabeth Mann, Xuhang Li, Francisco Nualart, Alejandro J. Yáñez
      Pages: 2437 - 2446
      Abstract: Diabetic nephropathy (DN) is characterized by interstitial inflammation and fibrosis, which is the result of chronic accumulation of extracellular matrix produced by activated fibroblasts in the renal tubulointerstitium. Renal proximal tubular epithelial cells (PTECs), through the process of epithelial‐to‐mesenchymal transition (EMT), are the source of fibroblasts within the interstitial space, and loss of E‐cadherin has shown to be one of the earliest steps in this event. Here, we studied the effect of the anti‐diabetic agent sodium tungstate (NaW) in the loss of E‐cadherin induced by transforming growth factor (TGF) β‐1, the best‐characterized in vitro EMT promoter, and serum from untreated or NaW‐treated diabetic rats in HK‐2 cell line, a model of human kidney PTEC. Our results showed that both TGFβ‐1 and serum from diabetic rat induced a similar reduction in E‐cadherin expression. However, E‐cadherin loss induced by TGFβ‐1 was not reversed by NaW, whereas sera from NaW‐treated rats were able to protect HK‐2 cells. Searching for soluble mediators of NaW effect, we compared secretion of TGFβ isoforms and vascular endothelial growth factor (VEGF)‐A, which have opposite actions on EMT. One millimolar NaW alone reduced secretion of both TGFβ‐1 and ‐2, and stimulated secretion of VEGF‐A after 48 h. However, these patterns of secretion were not observed after diabetic rat serum treatment, suggesting that protection from E‐cadherin loss by serum from NaW‐treated diabetic rats originates from an indirect rather than a direct effect of this salt on HK‐2 cells, via a mechanism independent of TGFβ and VEGF‐A functions. J. Cell. Physiol. 230: 2437–2446, 2015. © 2015 Wiley Periodicals, Inc.
      PubDate: 2015-06-23T17:18:38.606421-05:
      DOI: 10.1002/jcp.24974
       
  • A SMYD3 Small‐Molecule Inhibitor Impairing Cancer Cell Growth
    • Authors: Alessia Peserico; Aldo Germani, Paola Sanese, Armenio Jorge Barbosa, Valeria Di Virgilio, Raffaella Fittipaldi, Edoardo Fabini, Carlo Bertucci, Greta Varchi, Mary Pat Moyer, Giuseppina Caretti, Alberto Del Rio, Cristiano Simone
      Pages: 2447 - 2460
      Abstract: SMYD3 is a histone lysine methyltransferase that plays an important role in transcriptional activation as a member of an RNA polymerase complex, and its oncogenic role has been described in different cancer types. We studied the expression and activity of SMYD3 in a preclinical model of colorectal cancer (CRC) and found that it is strongly upregulated throughout tumorigenesis both at the mRNA and protein level. Our results also showed that RNAi‐mediated SMYD3 ablation impairs CRC cell proliferation indicating that SMYD3 is required for proper cancer cell growth. These data, together with the importance of lysine methyltransferases as a target for drug discovery, prompted us to carry out a virtual screening to identify new SMYD3 inhibitors by testing several candidate small molecules. Here we report that one of these compounds (BCI‐121) induces a significant reduction in SMYD3 activity both in vitro and in CRC cells, as suggested by the analysis of global H3K4me2/3 and H4K5me levels. Of note, the extent of cell growth inhibition by BCI‐121 was similar to that observed upon SMYD3 genetic ablation. Most of the results described above were obtained in CRC; however, when we extended our observations to tumor cell lines of different origin, we found that SMYD3 inhibitors are also effective in other cancer types, such as lung, pancreatic, prostate, and ovarian. These results represent the proof of principle that SMYD3 is a druggable target and suggest that new compounds capable of inhibiting its activity may prove useful as novel therapeutic agents in cancer treatment. J. Cell. Physiol. 230: 2447–2460, 2015. © 2015 Wiley Periodicals, Inc.
      PubDate: 2015-06-23T17:18:40.40041-05:0
      DOI: 10.1002/jcp.24975
       
  • Paracrine Factors Secreted by MSCs Promote Astrocyte Survival Associated
           With GFAP Downregulation After Ischemic Stroke via p38 MAPK and JNK
    • Authors: Weiyi Huang; Bingke Lv, Huijun Zeng, Dandan Shi, Yi Liu, Fanfan Chen, Feng Li, Xinghui Liu, Rong Zhu, Lei Yu, Xiaodan Jiang
      Pages: 2461 - 2475
      Abstract: Astrocytes are critical for ischemic stroke, and understanding their role in mesenchymal stem cell (MSC)‐mediated protection against ischemic injury is important. The paracrine capacity of MSCs has been proposed as the principal mechanism contributing to the protection and repair of brain tissue. In the present study, an in vitro oxygen–glucose deprivation (OGD) model was used to mimic ischemic injury. OGD‐induced astrocytes were reperfused with MSC‐conditioned medium (MSC‐CM) or co‐cultured with MSCs for 24 h to create an environment abundant in paracrine factors. The results indicated that both situations could protect astrocytes from apoptosis, increase cell metabolic activity, and reduce glial fibrillary acidic protein (GFAP) overexpression; however, the effects of co‐culturing with MSCs were more positive. Paracrine factors suppressed the activation of p38 MAPK, JNK, and their downstream targets p53 and STAT1. Inhibition of p38 MAPK, JNK, p53, and STAT1 attenuated astrocyte injury and/or GFAP upregulation. Activation of p38 MAPK and JNK suppressed the beneficial effects of paracrine factors, resulting in decreased survival and GFAP overexpression. These results suggest that paracrine factors inhibit p38 MAPK and JNK, and most likely by regulating their downstream targets, p53 and STAT1, to promote astrocyte survival associated with GFAP downregulation after ischemic stroke in vitro. J. Cell. Physiol. 230: 2461–2475, 2015. © 2015 Wiley Periodicals, Inc.
      PubDate: 2015-06-23T17:18:37.493492-05:
      DOI: 10.1002/jcp.24981
       
  • Gap Junctions Enhance the Antiproliferative Effect of
           MicroRNA‐124‐3p in Glioblastoma Cells
    • Authors: Suzhi Zhang; Liang Tao, Yuexia Peng, Lucy Liu, Xiaoting Hong, Yuan Zhang, Qin Wang
      Pages: 2476 - 2488
      Abstract: MicroRNA (miRNA) holds promise as a novel therapeutic tool for cancer treatment. However, the transfection efficiency of current delivery systems represents a bottleneck for clinical applications. Here, we demonstrate that gap junctions mediate an augmentative effect on the antiproliferation mediated by miR‐124‐3p in U87 and C6 glioblastoma cells. The functional inhibition of gap junctions using either siRNA or pharmacological inhibition eliminated the miR‐124‐3p‐mediated antiproliferation, whereas the enhancement of gap junctions with retinoic acid treatment augmented this miR‐124‐3p‐mediated antiproliferation. A similar effect was observed in glioblastoma xenograft models. More importantly, patch clamp and co‐culture assays demonstrated the transmission of miR‐124‐3p through gap junction channels into adjacent cells. In further exploring the impact of gap junction‐mediated transport of miR‐124‐3p on miR‐124‐3p target pathways, we found that miR‐124‐3p inhibited glioblastoma cell growth in part by decreasing the protein expression of cyclin‐dependent kinase 6, leading to cell cycle arrest at the G0/G1 phase; moreover, pharmacological regulation of gap junctions affected this cell cycle arrest. In conclusion, our results indicate that the “bystander” effects of functional gap junctions composed of connexin 43 enhance the antitumor effect of miR‐124‐3p in glioblastoma cells by transferring miR‐124‐3p to adjacent cells, thereby enhancing G0/G1 cell cycle arrest. These observations provide a new guiding strategy for the clinical application of miRNA therapy in tumor treatment. J. Cell. Physiol. 230: 2476–2488, 2015. © 2015 Wiley Periodicals, Inc.
      PubDate: 2015-06-23T17:18:45.189516-05:
      DOI: 10.1002/jcp.24982
       
  • Role of contraction duration in inducing fast‐to‐slow
           contractile and metabolic protein and functional changes in engineered
           muscle
    • Authors: Alastair Khodabukus; Leslie M. Baehr, Sue C. Bodine, Keith Baar
      Pages: 2489 - 2497
      Abstract: The role of factors such as frequency, contraction duration and active time in the adaptation to chronic low‐frequency electrical stimulation (CLFS) is widely disputed. In this study we explore the ability of contraction duration (0.6, 6, 60, and 600 sec) to induce a fast‐to‐slow shift in engineered muscle while using a stimulation frequency of 10 Hz and keeping active time constant at 60%. We found that all contraction durations induced similar slowing of time‐to‐peak tension. Despite similar increases in total myosin heavy (MHC) levels with stimulation, increasing contraction duration resulted in progressive decreases in total fast myosin. With contraction durations of 60 and 600 sec, MHC IIx levels decreased and MHC IIa levels increased. All contraction durations resulted in fast‐to‐slow shifts in TnT and TnC but increased both fast and slow TnI levels. Half‐relaxation slowed to a greater extent with contraction durations of 60 and 600 sec despite similar changes in the calcium sequestering proteins calsequestrin and parvalbumin and the calcium uptake protein SERCA. All CLFS groups resulted in greater fatigue resistance than control. Similar increases in GLUT4, mitochondrial enzymes (SDH and ATPsynthase), the fatty acid transporter CPT‐1, and the metabolic regulators PGC‐1α and MEF2 were found with all contraction durations. However, the mitochondrial enzymes cytochrome C and citrate synthase were increased to greater levels with contraction durations of 60 and 600 sec. These results demonstrate that contraction duration plays a pivotal role in dictating the level of CLFS‐induced contractile and metabolic adaptations in tissue‐engineered skeletal muscle. J. Cell. Physiol. 230: 2489–2497, 2015. © 2015 The
      Authors . Journal of Cellular Physiology Published by Wiley Periodicals, Inc.
      PubDate: 2015-06-23T17:18:56.774153-05:
      DOI: 10.1002/jcp.24985
       
  • Upregulation of miR‐155 in CD4+ T Cells Promoted Th1 Bias in
           Patients With Unstable Angina
    • Authors: Yanmei Chen; Lang Li, Yongguang Lu, Qiang Su, Yuhan Sun, Yang Liu, Dong Yang
      Pages: 2498 - 2509
      Abstract: An imbalance between T helper 1 (Th1) and T helper 2 (Th2) cells has been reported to increase plaque instability in patients with unstable angina (UA). MicroRNAs play a vital role in the differentiation of CD4+ T cells. However, the role of microRNAs in regulation of Th1/Th2 balance in UA remains unclear. In this study, we aimed to elucidate microRNA expression profiles of circulating CD4+ T cells in UA and to explore the function of microRNAs in the Th1/Th2 balance. A total of 53 patients with UA and 31 control subjects without coronary artery disease were enrolled. Microarray analysis of the microRNA expression profiles of CD4+ T cells revealed that miR‐155 was the most significantly upregulated microRNA of the 451 differentially expressed microRNAs. The upregulation of miR‐155 expression was positively correlated with the percentage of Th1 cells and interferon‐gamma (IFN‐γ) levels in patients with UA. In addition, overexpression of miR‐155 in human circulating CD4+ T cells promoted Th1 differentiation. Further studies identified IFN‐γ receptor alpha chain (IFN‐γ Rα) mRNA as a direct and functional target of miR‐155. A luciferase reporter assay verified that miR‐155 directly targeted IFN‐γ Rα mRNA. Small‐interfering RNA‐mediated knockdown of IFN‐γ Rα mRNA showed effects similar to those of ectopic miR‐155 expression. Thus, our study indicated that upregulation of miR‐155 in circulating CD4+ T cells in patients with UA promoted a shift in the Th1/Th2 balance toward Th1 dominance by repressing IFN‐γ Rα, which may subsequently enhance plaque instability. J. Cell. Physiol. 230: 2498–2509, 2015. © 2015 Wiley Periodicals, Inc.
      PubDate: 2015-06-23T17:18:43.658573-05:
      DOI: 10.1002/jcp.24987
       
  • Upregulation of Runt‐Related Transcription Factor‐2 Through
           CCAAT Enhancer Binding Protein‐β Signaling Pathway in
           Microglial BV‐2 Cells Exposed to ATP
    • Authors: Ryota Nakazato; Takeshi Takarada, Shinsuke Ikeno, Saki Nakamura, Takaya Kutsukake, Eiichi Hinoi, Yukio Yoneda
      Pages: 2510 - 2521
      Abstract: We have shown constitutive expression of the master regulator of osteoblastogenesis, runt‐related transcription factor‐2 (Runx2), by microglia cells outside bone. Here, we attempted to evaluate the pathological significance of Runx2 in microglial BV‐2 cells exposed to ATP at a high concentration. Marked upregulation of Runx2 transcript and protein expression was seen in cells exposed to 1 mM ATP for a period longer than 30 min without inducing cytotoxicity. The Runx2 upregulation by ATP was prevented by extracellular and intracellular Ca2+ chelators, while thapsigargin upregulated Runx2 expression alone without affecting the upregulation by ATP. A calmodulin antagonist prevented the upregulation by ATP, with calcineurin inhibitors being ineffective. Although ATP markedly increased nuclear levels of nuclear factor of activated T cell‐2 (NFAT2), Runx2 promoter activity was not simulated by the introduction of either NFAT1 or NFAT2, but facilitated by that of CCAAT enhancer binding protein‐α (C/EBPα), C/EBPβ and nuclear factor (erythroid‐derived 2)‐like‐2 (Nrf2). Exposure to ATP up‐regulated C/EBPβ and Nrf2, but not C/EBPα, expression, in addition to increasing nuclear levels of respective corresponding proteins. Runx2 upregulation by ATP was deteriorated by knockdown of C/EBPβ but not by that of Nrf2, however, while exposure to ATP up‐regulated matrix metalloproteinase‐13 (Mmp13) expression in a Runx2‐dependent manner. Overexpression of Runx2 up‐regulated Mmp13 expression with promoted incorporation of fluorescent beads into BV‐2 cells without ATP. These results suggest that extracellular ATP up‐regulates Runx2 expression through activation of the C/EBPβ signaling in a calmodulin‐dependent manner to play a pivotal role in phagocytosis in microglial BV‐2 cells. J. Cell. Physiol. 230: 2510–2521, 2015. © 2015 Wiley Periodicals, Inc.
      PubDate: 2015-06-23T17:18:56.032806-05:
      DOI: 10.1002/jcp.24988
       
  • Runx1 is associated with breast cancer progression in MMTV‐PyMT
           transgenic mice and its depletion in vitro inhibits migration and invasion
           
    • Authors: Gillian Browne; Hanna Taipaleenmäki, Nicole M. Bishop, Sharath C. Madasu, Leslie M. Shaw, Andre J. van Wijnen, Janet L. Stein, Gary S. Stein, Jane B. Lian
      Pages: 2522 - 2532
      Abstract: Runx1 is a transcription factor essential for definitive hematopoiesis, and genetic abnormalities in Runx1 cause leukemia. Runx1 is functionally promiscuous and acts as either an oncogene or tumor suppressor gene in certain epithelial cancers. Recent evidence suggests that Runx1 is an important factor in breast cancer, however, its role remains ambiguous. Here, we addressed whether Runx1 has a specific pathological role during breast cancer progression and show that Runx1 has an oncogenic function. We observed elevated Runx1 expression in a subset of human breast cancers. Furthermore, throughout the course of disease progression in a classical mouse model of breast cancer (i.e., the MMTV‐PyMT transgenic model), Runx1 expression increases in the primary site (mammary gland) and is further upregulated in tumors and distal lung metastatic lesions. Ex vivo studies using tumor epithelial cells derived from these mice express significantly higher levels of Runx1 than normal mammary epithelial cells. The tumor cells exhibit increased rates of migration and invasion, indicative of an aggressive cancer phenotype. Inhibition of Runx1 expression using RNA interference significantly abrogates these cancer‐relevant phenotypic characteristics. Importantly, our data establish that Runx1 contributes to murine mammary tumor development and malignancy and potentially represents a key disease‐promoting and prognostic factor in human breast cancer progression and metastasis. J. Cell. Physiol. 230: 2522–2532, 2015. © 2015 Wiley Periodicals, Inc.
      PubDate: 2015-06-23T17:18:46.265763-05:
      DOI: 10.1002/jcp.24989
       
  • Cell cycle gene expression networks discovered using systems biology:
           Significance in carcinogenesis
    • Authors: Robert E. Scott; Prachi N. Ghule, Janet L. Stein, Gary S. Stein
      Pages: 2533 - 2542
      Abstract: The early stages of carcinogenesis are linked to defects in the cell cycle. A series of cell cycle checkpoints are involved in this process. The G1/S checkpoint that serves to integrate the control of cell proliferation and differentiation is linked to carcinogenesis and the mitotic spindle checkpoint is associated with the development of chromosomal instability. This paper presents the outcome of systems biology studies designed to evaluate if networks of covariate cell cycle gene transcripts exist in proliferative mammalian tissues including mice, rats, and humans. The GeneNetwork website that contains numerous gene expression datasets from different species, sexes, and tissues represents the foundational resource for these studies (www.genenetwork.org). In addition, WebGestalt, a gene ontology tool, facilitated the identification of expression networks of genes that co‐vary with key cell cycle targets, especially Cdc20 and Plk1 (www.bioinfo.vanderbilt.edu/webgestalt). Cell cycle expression networks of such covariate mRNAs exist in multiple proliferative tissues including liver, lung, pituitary, adipose, and lymphoid tissues among others but not in brain or retina that have low proliferative potential. Sixty‐three covariate cell cycle gene transcripts (mRNAs) compose the average cell cycle network with P = e−13 to e−36. Cell cycle expression networks show species, sex and tissue variability, and they are enriched in mRNA transcripts associated with mitosis, many of which are associated with chromosomal instability. J. Cell. Physiol. 230: 2533–2542, 2015. © 2015 Wiley Periodicals, Inc.
      PubDate: 2015-06-23T17:18:48.31449-05:0
      DOI: 10.1002/jcp.24990
       
  • Lovastatin‐Mediated Changes in Human Tendon Cells
    • Authors: Maria Kuzma‐Kuzniarska; Hannah R. Cornell, Michael C. Moneke, Andrew J. Carr, Philippa A. Hulley
      Pages: 2543 - 2551
      Abstract: Statins are among the most widely prescribed drugs worldwide. Numerous studies have shown their beneficial effects in prevention of cardiovascular disease through cholesterol‐lowering and anti‐atherosclerotic properties. Although some statin patients may experience muscle‐related symptoms, severe side effects of statin therapy are rare, primarily due to extensive first‐pass metabolism in the liver. Skeletal muscles appear to be the main site of side effects; however, recently some statin‐related adverse effects have been described in tendon. The mechanism behind these side effects remains unknown. This is the first study that explores tendon‐specific effects of statins in human primary tenocytes. The cells were cultured with different concentrations of lovastatin for up to 1 week. No changes in cell viability or morphology were observed in tenocytes incubated with therapeutic doses. Short‐term exposure to lovastatin concentrations outside the therapeutic range had no effect on tenocyte viability; however, cell migration was reduced. Simvastatin and atorvastatin, two other drug family members, also reduced the migratory properties of the cells. Prolonged exposure to high concentrations of lovastatin induced changes in cytoskeleton leading to cell rounding and decreased levels of mRNA for matrix proteins, but increased BMP‐2 expression. Gap junctional communication was impaired but due to cell shape change and separation rather than direct gap junction inhibition. These effects were accompanied by inhibition of prenylation of Rap1a small GTPase. Collectively, we showed that statins in a dose‐dependent manner decrease migration of human tendon cells, alter their expression profile and impair the functional network, but do not inhibit gap junction function. J. Cell. Physiol. 230: 2543–2551, 2015. © 2015 The
      Authors . Journal of Cellular Physiology Published by Wiley Periodicals, Inc.
      PubDate: 2015-06-23T17:18:39.703176-05:
      DOI: 10.1002/jcp.25010
       
  • GRP78/Dna K Is a Target for Nexavar/Stivarga/Votrient in the Treatment of
           Human Malignancies, Viral Infections and Bacterial Diseases
    • Authors: Jane L. Roberts; Mehrad Tavallai, Aida Nourbakhsh, Abigail Fidanza, Tanya Cruz‐Luna, Elizabeth Smith, Paul Siembida, Pascale Plamondon, Kelly A. Cycon, Christopher D. Doern, Laurence Booth, Paul Dent
      Pages: 2552 - 2578
      Abstract: Prior tumor cell studies have shown that the drugs sorafenib (Nexavar) and regorafenib (Stivarga) reduce expression of the chaperone GRP78. Sorafenib/regorafenib and the multi‐kinase inhibitor pazopanib (Votrient) interacted with sildenafil (Viagra) to further rapidly reduce GRP78 levels in eukaryotes and as single agents to reduce Dna K levels in prokaryotes. Similar data were obtained in tumor cells in vitro and in drug‐treated mice for: HSP70, mitochondrial HSP70, HSP60, HSP56, HSP40, HSP10, and cyclophilin A. Prolonged ‘rafenib/sildenafil treatment killed tumor cells and also rapidly decreased the expression of: the drug efflux pumps ABCB1 and ABCG2; and NPC1 and NTCP, receptors for Ebola/Hepatitis A and B viruses, respectively. Pre‐treatment with the ‘Rafenib/sildenafil combination reduced expression of the Coxsackie and Adenovirus receptor in parallel with it also reducing the ability of a serotype 5 Adenovirus or Coxsackie virus B4 to infect and to reproduce. Sorafenib/pazopanib and sildenafil was much more potent than sorafenib/pazopanib as single agents at preventing Adenovirus, Mumps, Chikungunya, Dengue, Rabies, West Nile, Yellow Fever, and Enterovirus 71 infection and reproduction. ‘Rafenib drugs/pazopanib as single agents killed laboratory generated antibiotic resistant E. coli which was associated with reduced Dna K and Rec A expression. Marginally toxic doses of ‘Rafenib drugs/pazopanib restored antibiotic sensitivity in pan‐antibiotic resistant bacteria including multiple strains of blakpc Klebsiella pneumoniae. Thus, Dna K is an antibiotic target for sorafenib, and inhibition of GRP78/Dna K has therapeutic utility for cancer and for bacterial and viral infections. J. Cell. Physiol. 230: 2552–2578, 2015. © 2015 Wiley Periodicals, Inc.
      PubDate: 2015-06-23T17:18:50.894854-05:
      DOI: 10.1002/jcp.25014
       
  • Targeting HMGB1 Inhibits Ovarian Cancer Growth and Metastasis by
           Lentivirus‐Mediated RNA Interference
    • Authors: Chen J; Liu X, Zhang J, Zhao Y
      Pages: 2579 - 2579
      PubDate: 2015-06-23T17:18:39.435886-05:
      DOI: 10.1002/jcp.25027
       
 
 
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