Associations Of Methylation Status In The GATA6Promoter Region With Congential Heart Diseases

Objective:Explore the etiology and pathogenesis of congenital heart diseases(CHDs), and then take effective prevention and treatment measures, the purpose of thisstudy and the urgent need to solve the puzzles.Currently CHDs in the etiology andpathogenesis mechanisms have not been fully elucidated, some literature suggests thatGATA6gene mutation related with the occurrence of CHDs, studies have shown that thepresence of abnormal methylation of this gene in cancer pathogenesis, but its promoterregion methylation abnormalities associated with CHDs are no reports. This studycomparing CHDs with non CHDs children GATA6mRNA expression level andmethylation status of the promoter region, aimed at understanding the correlation of thepathogenesis of CHDs with GATA6aberrant methylation of the promoter region.Methods：Children with CHDs collected peripheral venous blood samples of90cases,blood samples of50cases of non-CHDs children,Total RNA extracted from peripheralblood,using real-time fluorescent quantitative PCR(qRT-PCR)technology,cycle threshold(Ct) comparison method for quantitative analysis for mRNA expression;DNA extractedfrom peripheral blood cells at the same time, the heavy sodium bisulfite modification,using Methylation specific PCR (MSP) to detect GATA6promoter regions CpG islandmethylation status; after using Bisulfite genomic sequencing PCR (BSP) method, designspecific primers cloning sequencing,GATA6were detected for each gene methylationstatus of CpG sites and the extent to further clarify the relationship between the two.Results：1、GATA6in the control group of non CHDs, the relative expression of CHDs setamount (x±s) were0.10±0.27,0.01±0.02, there are significant differences between thetwo groups (P <0.05).2、 GATA6promoter in the control group of non-CHDs,CHDs groups weremethylated phenomenon occurs, methylation-positive rate was47%(17/36),91%(69/76), the former was significantly lower than the latter, the two groups have statisticallysignificant difference (P <0.05).3、GATA6in the control group of non-CHDs, CHDs group showed some degree ofabnormal methylation,BSP method positive rate was50%(5/10),75%(15/20), nosignificant difference between the two groups (P>0.05).Conclusions：1、GATA6mRNAreduce the expression of CHDs group, suggesting the formation of anew exception may GATA6expression of genes involved in CHDs.2、GATA6promoter methylation status of the region there is a significant differencebetween children with CHDs with normal control group, suggesting that GATA6genepromoter methylation abnormalities associated with CHDs occur.