title: The role of nuclear factor of activated T cells during phorbol myristate acetate-induced cardiac differentiation of mesenchymal stem cells.
abstract: BACKGROUND
  We previously reported that phorbol 12-myristate 13-acetate (PMA) treatment can induce the cardiac differentiation of mesenchymal stem cells (MSCs). In the present study, we investigated how PMA induces cardiac differentiation of MSCs, focusing on its effect on the transcription factors responsible for increased cardiac marker gene expression.
  
  
  METHODS
  Human MSCs (hMSCs) were treated with 1 μM PMA for 9 days. The expression of MSC markers and cardiac markers in the PMA-treated hMSC, as well as the nuclear translocation of transcription factors, nuclear factor of activated T cells (NFAT), and myogenic differentiation 1 (MyoD), was examined. Transcriptional activity of NFAT was examined by utilizing a green fluorescent protein (GFP) vector containing NFAT motif of human interleukin-2 promoter. The effect of PMA on the expression of key cell cycle regulators was examined.
  
  
  RESULTS
  PMA induces the transcriptional activity of NFAT and MyoD, which have been associated with increased expression of cardiac troponin T (cTnT) and myosin heavy chain (MHC), respectively. Our data suggested that protein kinase C (PKC) mediates the effect of PMA on NFAT activation. Furthermore, PMA treatment increased cell-cycle regulator p27(kip1) expression, suggesting that PMA triggers the cardiac differentiation program in MSCs by regulating key transcription factors and cell cycle regulators.
  
  
  CONCLUSIONS
  The results of this study demonstrate the importance of NFAT activation during PMA-induced MSC differentiation and help us to better understand the underlying mechanisms of small molecule-mediated MSC differentiation so that we can develop a strategy for synthesizing novel and improved differentiation-inducing small molecules.
authors.affiliation: Brain Korea 21 PLUS Project for Medical Science, Yonsei University, Seoul, South Korea.
authors.forename: Hyang-Hee
authors.initials: HH
authors.lastname: Seo
authors.affiliation: Department of Integrated Omics for Biomedical Sciences, Yonsei University, Seoul, South Korea.
authors.forename: Chang Youn
authors.initials: CY
authors.lastname: Lee
authors.affiliation: Brain Korea 21 PLUS Project for Medical Science, Yonsei University, Seoul, South Korea.
authors.forename: Jiyun
authors.initials: J
authors.lastname: Lee
authors.affiliation: Institute for Bio-medical Convergence, Catholic Kwandong University, Incheon, South Korea.
authors.forename: Soyeon
authors.initials: S
authors.lastname: Lim
authors.affiliation: Institute for Bio-medical Convergence, Catholic Kwandong University, Incheon, South Korea.
authors.forename: Eunhyun
authors.initials: E
authors.lastname: Choi
authors.affiliation: Cellbiocontrol Laboratory, Department of Medical Engineering, Yonsei University College of Medicine, Seoul, South Korea.
authors.forename: Jong-Chul
authors.initials: JC
authors.lastname: Park
authors.affiliation: Institute for Bio-medical Convergence, Catholic Kwandong University, Incheon, South Korea. sam1017@ish.ac.kr.|Department of Biomedical Sciences, College of Medicine, Catholic Kwandong University, Gangneung, Gangwon-do, South Korea. sam1017@ish.ac.kr.
authors.forename: Seahyoung
authors.initials: S
authors.lastname: Lee
authors.affiliation: Institute for Bio-medical Convergence, Catholic Kwandong University, Incheon, South Korea. kchwang@cku.ac.kr.|Department of Biomedical Sciences, College of Medicine, Catholic Kwandong University, Gangneung, Gangwon-do, South Korea. kchwang@cku.ac.kr.
authors.forename: Ki-Chul
authors.initials: KC
authors.lastname: Hwang
mesh_terms: D002453:Cell Cycle / Q000187:drug effects; D002454:Cell Differentiation / Q000187:drug effects; D050760:Cyclin-Dependent Kinase Inhibitor p27 / Q000235:genetics / Q000378:metabolism; D005786:Gene Expression Regulation; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008213:Lymphocyte Activation / Q000187:drug effects; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D017570:MyoD Protein / Q000235:genetics / Q000378:metabolism; D032383:Myocytes, Cardiac / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D018995:Myosin Heavy Chains / Q000235:genetics / Q000378:metabolism; D050778:NFATC Transcription Factors / Q000235:genetics* / Q000378:metabolism; D061251:Primary Cell Culture; D011493:Protein Kinase C / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D013601:T-Lymphocytes / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology*; D014158:Transcription, Genetic; D020107:Troponin T / Q000235:genetics / Q000378:metabolism
chemical_list: C436291:CDKN1B protein, human; D017570:MyoD Protein; C057363:MyoD1 myogenic differentiation protein; D050778:NFATC Transcription Factors; D020107:Troponin T; D049452:Green Fluorescent Proteins; D050760:Cyclin-Dependent Kinase Inhibitor p27; D011493:Protein Kinase C; D018995:Myosin Heavy Chains; D013755:Tetradecanoylphorbol Acetate
keywords: Cardiac differentiation; MSC; NFAT; Phorbol myristate acetate; Small molecule