Open Access Research article

DNA methylation status of NKX2-5, GATA4 and HAND1 in patients with tetralogy of fallot

Wei Sheng13, Yanyan Qian2, Huijun Wang13, Xiaojing Ma13, Ping Zhang3, Lianwei Diao5, Quan An5, Long Chen4, Duan Ma23* and Guoying Huang13*

Author Affiliations

1 Children Hospital of Fudan University, Shanghai 201102, China

2 Key Laboratory of Molecular Medicine, Ministry of Education, Department of Biochemistry and Molecular Biology, Institutes of Biomedical Sciences, Shanghai Medical College, Fudan University, Shanghai 20003, China

3 Shanghai Key Laboratory of Prevention and Intervention of Birth Defects, Shanghai 201102, China

4 Department of Forensic Medicine, Fudan University, Shanghai 20003, China

5 Wuhe Chinese Medicine Hospital, Wuhe 233300, China

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BMC Medical Genomics 2013, 6:46  doi:10.1186/1755-8794-6-46

Published: 1 November 2013



NKX2-5, GATA4 and HAND1 are essential for heart development, however, little is known regarding their epigenetic regulation in the pathogenesis of tetralogy of fallot (TOF).


Methylation levels were measured in three regions of NKX2-5 (M1: -1596 bp ~ -1374 bp, M2: -159 bp ~ 217 bp and M3: 1058 bp ~ 1524 bp), one region of GATA4 (M: -392 bp ~ 107 bp) and three regions of HAND1 (M1: -887 bp ~ -414 bp, M2: -436 bp ~ 2 bp and M3: 37 bp ~ 398 bp) using the Sequenom MassARRAY platform. QRT-PCR was used to analyze NKX2-5 and HAND1 mRNA levels in the right ventricular myocardium of TOF patients.


TOF patients had a significantly higher NKX2-5_M3 median methylation level than controls (41.65% vs. 22.18%; p = 0.0074; interquartile range [IQR]: 30.46%–53.35%, N = 30 and 20.07%–24.31%, N = 5; respectively). The HAND1_M1 median methylation level was also significantly higher in TOF patients than controls (30.05% vs. 17.54%; p = 0.0054; IQR: 20.77%–40.89%, N = 30 and IQR: 14.69%–20.64%; N = 6; respectively). The methylation statuses of NKX2-5_M1, NKX2-5_M2, GATA4_M, HAND1_M2 or HAND1_M3 were not significantly different in TOF patients compared to controls. The methylation values for NKX2-5_M3 were negatively correlated with mRNA levels (r = - 0.463, p = 0.010, N = 30) and there was a significant association between HAND1_M1 methylation status and mRNA levels (r = - 0.524, p = 0.003, N = 30) in TOF patients.


Aberrant methylation statuses of the NKX2-5 gene body and HAND1 promoter regions are associated with the regulation of gene transcription in TOF patients and may play an important role in the pathogenesis of TOF.

DNA methylation; NKX2-5; GATA4 and HAND1 genes; Tetralogy of fallot