title: [Experimental studies on the role of GATA4 in the endocardial cushions development].
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 Hela(GATA4), Hela(H436Y), Hela(Null) 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 GATA4(H436Y) 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.
authors.affiliation: Department of pediatric, Anhui Medical University Affiliated to Anhui Provincial Hospital, Hefei 230001, China.
authors.forename: Yanli
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Department of pediatric, Anhui Medical University Affiliated to Anhui Provincial Hospital, Hefei 230001, China; Email: chenmingwu01@163.com.
authors.forename: Mingwu
authors.initials: M
authors.lastname: Chen
authors.affiliation: Department of pediatric, Anhui Medical University Affiliated to Anhui Provincial Hospital, Hefei 230001, China.
authors.forename: Ye
authors.initials: Y
authors.lastname: Chen
authors.affiliation: Department of pediatric, Anhui Medical University Affiliated to Anhui Provincial Hospital, Hefei 230001, China.
authors.forename: Shujun
authors.initials: S
authors.lastname: Hong
mesh_terms: D053674:Aggrecans / Q000378:metabolism; D051792:Basic Helix-Loop-Helix Proteins / Q000378:metabolism; D015536:Down-Regulation; D054089:Endocardial Cushions / Q000196:embryology*; D050991:GATA4 Transcription Factor / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D005822:Genetic Vectors; D006367:HeLa Cells; D006801:Humans; D012333:RNA, Messenger; D055758:SOX9 Transcription Factor / Q000378:metabolism; D019063:Tenascin / Q000378:metabolism; D014162:Transfection
chemical_list: D053674:Aggrecans; D051792:Basic Helix-Loop-Helix Proteins; D050991:GATA4 Transcription Factor; D012333:RNA, Messenger; D055758:SOX9 Transcription Factor; D019063:Tenascin; C508319:ACAN protein, human; C484170:GATA4 protein, human; C093504:SCX protein, human; C527932:SOX9 protein, human