DNA hypermethylation of the NOX5 gene in fetal ventricular septal defect

Zhu,Chun; Yu,Zhang-Bin; Chen,Xiao-Hui; Ji,Chen-Bo; Qian,Ling-Mei; Han,Shu-Ping

doi:10.3892/etm.2011.294

DNA hypermethylation of the NOX5 gene in fetal ventricular septal defect

Authors:
- Chun Zhu
- Zhang-Bin Yu
- Xiao-Hui Chen
- Chen-Bo Ji
- Ling-Mei Qian
- Shu-Ping Han
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Affiliations: Department of Pediatrics, Nanjing Maternal and Child Health Hospital of Nanjing Medical University, Nanjing 210004, P.R. China, Institute of Pediatrics, Nanjing Medical University, Nanjing 210029, P.R. China, Department of Cardiology, The First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, P.R. China
Published online on: June 24, 2011 https://doi.org/10.3892/etm.2011.294
Pages: 1011-1015

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Abstract

Ventricular septal defect (VSD) is the most comon form of congenital heart disease (CHD). DNA hypermethylation analysis may provide an insight into the molecular features and pathogenesis of this heart disease. Although aberrant DNA hypermethylation is implicated in the pathophysiology of this heart disease, only a limited number of genes are known to be epigenetically altered in VSD. We previously identified regulation of the NOX5 gene by hypermethylation in VSD fetuses by promoter methylation microarrays. This study was designed to detect the expression of NOX5 mRNA in VSD and normal fetuses. We also verified the results of promoter methylation microarrays by methylation-specific PCR. DNA extraction and nested methylation-specific PCR were performed on myocardial tissue samples from 21 VSD and 15 normal fetuses. The primers specific for methylated vs. unmethylated DNA were designed and amplified by nested PCR. The products were visualized on agarose gel. Hypermethylation of the NOX5 promoter was more frequent in VSD fetuses (66.67%) than in normal fetuses (20%). There was a significant concordance between NOX5 methylation and a decrease in its mRNA expression. Taken together, our results demonstrate that hypermethylation of the NOX5 gene may be involved in the pathogenesis of VSD.

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