title: [Molecular mechanism of cardiac differentiation in P19 embryonal carcinoma cells regulated by Foxa2].
abstract: OBJECTIVE
  To investigate the involvement of transcription factor Foxa2 in cardiac differentiation in P19 embryonal carcinoma cells and its molecular mechanism.
  
  
  METHODS
  P19 cells were induced to differentiate into cardiomyocytes by adding dimethyl sulfoxide (DMSO) into the culture medium of their embryoid bodies (EBs). The mRNA levels of pluripotency markers of embryonic pluripotent stem cells, cardiac differentiation related genes, and Foxa2 in the cell samples at different time points of cardiac differentiation were detected by reverse transcription PCR (RT-PCR). Differentiated and mature cardiomyocytes were identified by immunofluorescence. Eukaryotic expression plasmid pCMV-rFoxa2 (rat Foxa2) was transfected into P19 cells, and clonal populations of P19 cells that stably expressed green fluorescence protein (GFP)-rFoxa2 were isolated to enhance the expression levels of Foxa2 in P19 cells. The mRNA and protein levels of pluripotency markers and cardiac differentiation related genes in the above cell samples were detected by RT-PCR and Western blot. The mRNA levels of cardiac differentiation related genes in EBs differentiation system were also examined.
  
  
  RESULTS
  P19 cells differentiated into cardiomyocytes in the presence of DMSO, accompanied by stimulated expression of Foxa2. Transfection of pCMV-rFoxa2 plasmids into P19 cells upregulated rFoxa2 expression transiently and activated the transcription of its downstream cardiac inducer Cerberus1 (Cer1). The expression of pluripotency marker Nanog was suppressed and the expression of cardiac inducer Sonic Hedgehog (Shh) was elevated in GFP-rFoxa2 P19 cells. The expression of Cer1 and cardiac muscle marker actin, alpha cardiac muscle 1 (Actc1) was upregulated in EBs of GFP-rFoxa2 P19 cells.
  
  
  CONCLUSIONS
  Foxa2 participates in cardiac differentiation in P19 embryonal carcinoma cells. Foxa2 may inhibit Nanog expression and stimulate the expression of Cer1 and Shh directly during cardiac differentiation in P19 cells in the presence of DMSO.
authors: Department of Biomedical Engineering, Hunan University, Changsha, China. zhuh6477@126.com
authors: Hong
authors: H
authors: Zhu
authors: Zhen
authors: Z
authors: Zhang
authors: Yi
authors: Y
authors: Liu
authors: Yan
authors: Y
authors: Chen
authors: Yongjun
authors: Y
authors: Tan
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects*; D002460:Cell Line; D016207:Cytokines; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D054278:Embryonal Carcinoma Stem Cells / Q000473:pathology*; D053823:Hedgehog Proteins / Q000378:metabolism; D051542:Hepatocyte Nuclear Factor 3-beta / Q000502:physiology*; D018398:Homeodomain Proteins / Q000378:metabolism; D051379:Mice; D032383:Myocytes, Cardiac / Q000166:cytology*; D000071317:Nanog Homeobox Protein; D011506:Proteins / Q000378:metabolism; D014162:Transfection
chemical_list: C110586:Cer1 protein, mouse; D016207:Cytokines; C496503:Foxa2 protein, mouse; D053823:Hedgehog Proteins; D018398:Homeodomain Proteins; D000071317:Nanog Homeobox Protein; C475540:Nanog protein, mouse; D011506:Proteins; C465570:Shh protein, mouse; D051542:Hepatocyte Nuclear Factor 3-beta; D004121:Dimethyl Sulfoxide