High CpG island methylation of p16 gene and loss of p16 protein expression associate with the development and progression of tetralogy of Fallot

Abstract

We examined CpG island methylation in p16 gene and its effect on p16 protein expression in tetralogy of Fallot (ToF) patients to explore its potential implications in the development and progression of ToF. The study subjects consisted of 75 healthy controls and 63 ToF patients recruited at Linyi People's Hospital between January 2012 and June 2014. The 4 mL of peripheral venous blood of each subject was obtained and saved in ethylene diamine tetraacetic acid (EDTA) tubes. Methylation-specific polymerase chain reaction (MSP) was employed to detect CpG island methylation in p16 promoter region andWestern blotting was used to detect p16 expression of all subjects. Real-time fluorescence quantitative polymerase chain reaction (FQ-PCR) was performed to test p16 mRNA expression. The results showed that p16-methylation rates in ToF group were significantly higher than the control group (ToF group, 58.73%; control group, 13.33%; P < 0.001). Remarkably, Western blotting and FQ-PCR results derived from RVOT revealed that p16 protein expression was significantly lower in ToF group compared tothe control group (0.76 ± 0.21 versus 2.31 ± 0.35; P < 0.001), and p16 gene expression was also markedly decreased in ToF group (1.212 ± 0.152 versus 1.346 ± 0.191, P < 0.001). Additionally, our analysis suggested that CpG island methylation in p16 promoters in ToF patients was negatively correlated with p16 protein and gene expression (both P < 0.05). Our study reports that high CpG island methylation of p16 gene and loss of p16 protein expression associate with the development and progression of ToF, which may have significant therapeutic applications for ToF.

Indole Derivative Interacts with Estrogen Receptor Beta and Inhibits Human Ovarian Cancer Cell Growth

“…CDKN2A is located on chromosome 9p21 and plays an important role in cell cycle regulation by slowing cells progression from G1 to S phase [ 32 , 33 ]. It has become clear that the expression of CDKN2A is reduced by DNA methylation [ 18 , 34 , 35 ]. In addition, CDKN2A inactivation upregulates RB, stimulating the cyclin-dependent kinases (CDKs) and the RB pathway, which leads to malfunction of cell proliferation and apoptosis, thereby further facilitating carcinogenesis [ 36 ].…”

Comparison of DNA methylation patterns across tissue types in infants with tetralogy of Fallot

“…Prior studies have shown DNA methylation abnormalities in medically-terminated fetal heart tissues in both syndromic and nonsyndromic forms of CHD, including TOF, compared to similar age non-CHD heart controls (Serra-Juhe et al, 2015). Previous studies also observed aberrant DNA methylation patterns in cardiac development genes in heart (Gong et al, 2019;Grunert et al, 2016;Sheng et al, 2013Sheng et al, , 2016Xiaodi et al, 2020;Yuan et al, 2014;Zhu et al, 2020) and blood samples (Chang et al, 2021;Gao, Zhang, & Zhang, 2016;Radhakrishna et al, 2018;Serra-Juhe et al, 2015) in infants with TOF, but it is not known if these aberrant DNA methylation patterns are conserved across tissue types. Also, the degree of variation in DNA methylation profiles for myocardial, blood, and buccal cells across different TOF patients is not known.…”

Quantitative assessment of aberrant <em>P16^INK4a</em> methylation in ovarian cancer: a meta-analysis based on literature and TCGA datasets

“…It has become clear that the expression of P16 is reduced by DNA methylation 13–15. Also, P16 INK4a inactivation upregulates retinoblastoma (RB) protein by stimulating the cyclin-dependent kinases (CDKs) and RB pathway, which leads to dysfunction of cell proliferation and apoptosis, thereby further facilitating carcinogenesis 16.…”