title: [Involvement of ERK5 and JNK in the BMP9-induced differentiation of C3H10T1/2 cells into cardiomyocyte-like cells].
abstract: OBJECTIVE
  To investigate the roles of extracellular signal-regulated kinase 5 (ERK5) and c-Jun N-terminal kinase (JNK) in the differentiation of C3H10T1/2 cells into cardiomyocyte-like cells induced by bone morphogenetic protein 9 (BMP9).
  
  
  METHODS
  BMP9 gene was imported into C3H10T1/2 cells by recombinant adenovirus. Western blotting was used to detect the actived levels of ERK5 and JNK after cultivated with BMP9 and different concentrations of ERK5 specific inhibitor BIX02189 or JNK specific inhibitor SP600125; real-time quantitative PCR (qRT-PCR) was performed to analyze the expressions of myocardium specific genes GATA binding protein 4 (GATA4), myocyte enhancer factor 2C (MEF2C) after one week induced by BMP9; Western blotting was conducted to measure the expressions of myocardium specific proteins connexin 43 (CX43), cardiac troponin T (cTnT) after three weeks induced by BMP9, and immunofluorescence to observe the positions of CX43 and cTnT in the cells.
  
  
  RESULTS
  In the case of transfection efficiency up to 50%, BMP9 exceedingly activated ERK5 and JNK, and significantly increased their phosphorylation level (P<0.05). After BIX02189 inhibited the activity of ERK5, the expression levels of myocardial differentiation markers MEF2C, GATA4, CX43, cTnT of C3H10T1/2 cells were significantly suppressed (P<0.05); JNK specific inhibitor SP600125 also inhibited the expression levels of MEF2C, GATA4, CX43, cTnT, but the inhibition on MEF2C and GATA4 was not as notable as that of BIX02189 (P<0.05).
  
  
  CONCLUSIONS
  The excessive activation of ERK5 and JNK plays an important role in the differentiation of C3H10T1/2 cells into cardiomyocyte-like cells induced by BMP9.
authors.affiliation: Ministry of Education Key Laboratory of Child Development and Disorders, Key Laboratory of Pediatrics in Chongqing, Chongqing International Science and Technology Cooperation Center for Child Development with Disorders, Children's Hospital of Chongqing Medical University, Chongqing 400014, China.
authors.forename: Xuejing
authors.initials: X
authors.lastname: Geng
authors.affiliation: Ministry of Education Key Laboratory of Child Development and Disorders, Key Laboratory of Pediatrics in Chongqing, Chongqing International Science and Technology Cooperation Center for Child Development with Disorders, Children's Hospital of Chongqing Medical University, Chongqing 400014, China.
authors.forename: Shuxia
authors.initials: S
authors.lastname: Liu
authors.affiliation: Department of Cardiology, Children's Hospital of Chongqing Medical University, Chongqing 400014, China.
authors.forename: Yuan
authors.initials: Y
authors.lastname: Chen
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D000814:Aniline Compounds / Q000494:pharmacology; D000818:Animals; D000873:Anthracenes / Q000494:pharmacology; D015153:Blotting, Western; D002454:Cell Differentiation; D002460:Cell Line; D018031:Connexin 43 / Q000378:metabolism; D050991:GATA4 Transcription Factor / Q000235:genetics; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D055427:Growth Differentiation Factor 2 / Q000235:genetics / Q000378:metabolism*; D007211:Indoles / Q000494:pharmacology; D048031:JNK Mitogen-Activated Protein Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D064326:MEF2 Transcription Factors / Q000235:genetics; D051379:Mice; D008856:Microscopy, Fluorescence; D048054:Mitogen-Activated Protein Kinase 7 / Q000037:antagonists & inhibitors / Q000378:metabolism*; D032383:Myocytes, Cardiac / Q000166:cytology / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection / Q000379:methods; D020107:Troponin T / Q000378:metabolism
chemical_list: D000814:Aniline Compounds; D000873:Anthracenes; C533207:BIX 02189; D018031:Connexin 43; D050991:GATA4 Transcription Factor; C484171:Gata4 protein, mouse; C526108:Gdf2 protein, mouse; D055427:Growth Differentiation Factor 2; D007211:Indoles; D064326:MEF2 Transcription Factors; C578041:Mef2c protein, mouse; D020107:Troponin T; D049452:Green Fluorescent Proteins; C432165:pyrazolanthrone; D048031:JNK Mitogen-Activated Protein Kinases; D048054:Mitogen-Activated Protein Kinase 7