Abstract.To identify and provide a global assessment of dna methylation in fetal ventricular septal defect (VSd), genomic dna extracted from fetal myocardial tissue samples with VSd (n=21) and from normal fetal myocardial tissue samples (n=15) was analyzed for gene methylation using array-based technology. Furthermore, the Kiaa0310, raB43, SiVa1 and ndrG2 genes were randomly selected for validation analysis using methylation-specific PCR. Our results revealed that 70 and 85 genes were regulated by hypermethylation and hypomethylation, respectively, in VSd. different clusters of genes were associated with functions including embryo development, signal transduction, cell apoptosis and cell proliferation. In conclusion, this study identified a set of candidate genes whose expression is regulated by dna methylation in fetal VSd.
Introductioncongenital heart disease (cHd) is the most common type of developmental defect, occurring in almost 1% of all neonates (1). Ventricular septal defect (VSd) is the most commonly recognized cHd (2). VSd may exist alone or as an integral part of complex cHd (3). VSd is a multifactorial complex disease, in which genetic and environmental factors play important roles. despite the availability of several surgical techniques to treat VSd, the exact molecular mechanism of this type of cHd remains unclear.dna methylation has been widely recognized as a potent mechanism for silencing gene expression and maintaining genome stability (4). dna methylation is the predominant epigenetic alteration occurring in mammalian genomes, and plays a critical functional role in development, differentiation and disease (5). Previous studies have revealed that during embryonic development, the mammalian genome undergoes profound reprogramming of dna methylation patterns in the germ and early pre-implantation embryos (6). Furthermore, the different prototypes of genes in each cell, tissue and organ are thought to be regulated by dna methylation even during early development (7).The recent advent of array-based techniques offers the opportunity for more comprehensive DNA methylation profiling (8). Comparing the DNA methylation profiles of myocardial tissue samples from VSd and normal fetuses, we provide novel information for identifying gene methylation that may be implicated in the pathological consequences of VSd.
Materials and methodsTissue samples. Fetal myocardial tissue samples were obtained from nanjing Maternal and child Health Hospital. Myocardial tissue samples from 21 VSd and 15 normal fetuses at 26 weeks of gestation were obtained during surgery for pregnancy termination owing to trauma of the pregnant women. all samples were collected with the approval of the appropriate institute ethics committee, and written consent was provided by each pregnant woman and her family. The speciments were immediately snap frozen in liquid nitrogen and then stored at -80˚C until analysis.DNA methylation profiling by methylated DNA immunoprecipitation. The methylation profiling by methylated DNA immunoprecipitatio...