Detection of cardiac development gene after RNA interference for histone acetylation to MSCs induced by 5-azacytidine
- VernacularTitle:5-aza诱导干细胞经乙酰化特异RNAi后心肌细胞发育相关基因检测
- Author:
Jing ZHU
- Publication Type:Journal Article
- Keywords:
mesenchymal stem cell;
myocardial cell;
acetylation;
RNA interference
- From:Journal of Third Military Medical University
1988;0(06):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct 7 shRNA plasmids targeted to Gcn5 and to explore their role on histone acetylation in regulating MSCs differentiation into cardiomyocytes.Methods MSCs were isolated and passaged.Seven shRNA plasmids were constructed by pGenesil-1(4.9 kb) vector with GFP gene.MSCs were treated with 10 ?mol/L 5-azacytidine respectively for 24 h,2,4 and 7 d and transfected with the plasmids via lipofectamine~(TM)2000.Gcn5,as the marker of the acetylation,and GATA4,as the marker of cardiomyocytes,were detected in MSCs during differentiation with or without shRNA plasmids transfection by RT-PCR.Results MSCs were fibroblast-like but changed morphologically after induction.Seven Gcn5 shRNA plasmids were constructed and identified by restriction endonucleases PstⅠ/SalⅠand DNA sequencing.The expression of Gcn5 and GATA4 could be detected in MSCs without shRNA plasmids tranfection,but no expression after shRNA plasmids transfection.Conclusion Gcn5 plays a very important role in MSCs differentiation into cardiomyocytes,which lays the basis for further research of the importance of acetylation in the regulation of cell differentiation.
