Experimental studies on the role of GATA4 in the endocardial cushions development
10.3760/cma.j.issn.0253-3758.2015.05.015
- VernacularTitle:GATA4基因对心内膜垫发育中相关蛋白表达作用的实验研究
- Author:
Yanli ZHANG
1
;
Mingwu CHEN
;
Ye CHEN
;
Shujun HONG
Author Information
1. 安徽医科大学附属安徽省立医院儿科
- Keywords:
Endocardial cushion defects;
GATA4 transcription factor;
Mutation
- From:
Chinese Journal of Cardiology
2015;43(5):437-441
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the role of GATA4 gene in the endocardial cushions development.Methods Target gene eukaryote expression vectors were constructed by pcDNA3.1 (-) vector plasmid,and were identified by DNA sequence analysis.Recombinant plasmids were transfected into Hela cells with lipofectamine 2000,meanwhile Hela cells transfected with empty vector or those without transfection served as transfection control group and blank control group,respectively.Real-time PCR and Western blot were performed to detect the relative expression of mRNA and protein of transcription factors GATA4,Sox9,Scleraxis and ECM proteins Aggrecan,Tenascin in each group.Results The relative mRNA expression of GATA4 in experimental group was significantly higher than in transfection control group and blank control group.GATA4 mRNA expression in HelaGATA4、Hela436Y 、HelaNull and Hela group was 310.83 ± 2.39,146.35 ± 1.74,0.94 ± 0.32,1.00 ± 0.28,respectively (F =72.508,P ＜ 0.05).Western blot results were consistent with the results obtained by qRT-PCR.The relative mRNA and protein expressions of Sox9,Scleraxis,Aggrecan and Tenascin in both experimental groups were significantly higher than that in transfection control group and blank control group (P ＜ 0.05),and above gene expressions were significantly downregulated in GATA4H436Y group,while they were similar between transfection control group and blank control group (all P ＞ 0.05).Conclusions GATA4 H436Y mutation reduces it's transcriptional activation,which might serve as a theoretical framework to demonstrate the roles of GATA4 gene in endocardial cushion development.
