title: [β-catenin is involved in BMP9-induced differentiation of C3H10T1/2 cells into cardiomyocyte-like cells].
abstract: Objective To investigate the role of β-catenin in the differentiation of C3H10T1/2 cells into cardiomyocyte-like cells induced by bone morphogenetic protein 9 (BMP9). Methods C3H10T1/2 cells were transfected with the recombinant adenovirus expressing BMP9 (Ad-BMP9) and differentiated into cardiomyocytes in vitro for up to 21 days. The activated level of β-catenin in the cells after cultivated with Ad-BMP9 and different concentrations of β-catenin specific inhibitor XAV-939 was detected by Western blotting. Real-time quantitative PCR (qRT-PCR) was performed to evaluate the expressions of cardiac specific gene myocyte enhancer factor 2C (MEF2C) and GATA binding protein 4 (GATA4) in the cells one week after induced by Ad-BMP9 and different concentrations of XAV-939. Three weeks after the transfection, the expressions of connexin 43 (Cx43) and cardiac troponin T (cTnT) were analyzed by Western blotting, and the location of Cx43 in the cells was observed by immunofluorescence technique. Results While the transfection was successful in 50% of the cells positive for BMP9, β-catenin was excessively activated and their phosphorylation level remarkably increased. After XAV-939 inhibited the activity of β-catenin, the expressions of MEF2C, GATA4, Cx43, cTnT in C3H10T1/2 cells induced by BMP9 were significantly suppressed. Conclusion The β-catenin can be activated by BMP9 and the activation of β-catenin 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, Chongqing Key Laboratory of Pediatrics, Chongqing International Science and Technology Cooperation Center for Child Development and Disorders, Chongqing 400014, China.
authors.forename: Songhua
authors.initials: S
authors.lastname: Fang
authors.affiliation: Ministry-of-Education Key Laboratory of Child Development and Disorders, Chongqing Key Laboratory of Pediatrics, Chongqing International Science and Technology Cooperation Center for Child Development and Disorders, Chongqing 400014, China.
authors.forename: Shuxia
authors.initials: S
authors.lastname: Liu
authors.affiliation: Department of Cardiology, Children's Hospital, Chongqing Medical University, Chongqing 400014, China. *Corresponding author, E-mail: cy111938@126.com.
authors.forename: Yuan
authors.initials: Y
authors.lastname: Chen
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002454:Cell Differentiation / Q000235:genetics*; D002460:Cell Line; D018031:Connexin 43 / Q000235:genetics / Q000378:metabolism; D050991:GATA4 Transcription Factor / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression / Q000187:drug effects; D055427:Growth Differentiation Factor 2 / Q000235:genetics* / Q000378:metabolism; D006575:Heterocyclic Compounds, 3-Ring / Q000494:pharmacology; D064326:MEF2 Transcription Factors / Q000235:genetics / Q000378:metabolism; D051379:Mice; D032383:Myocytes, Cardiac / Q000166:cytology / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013997:Time Factors; D014162:Transfection; D020107:Troponin T / Q000235:genetics / Q000378:metabolism; D051176:beta Catenin / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism
chemical_list: D018031:Connexin 43; D050991:GATA4 Transcription Factor; C484171:Gata4 protein, mouse; C526108:Gdf2 protein, mouse; D055427:Growth Differentiation Factor 2; D006575:Heterocyclic Compounds, 3-Ring; D064326:MEF2 Transcription Factors; D020107:Troponin T; C544261:XAV939; D051176:beta Catenin