Abstract
Hepatitis B virus (HBV) is a hepatotropic virus causing hepatitis, cirrhosis and hepatocellular carcinoma (HCC). The methylation status of the HBV DNA in its different forms can potentially provide insight into the pathogenesis of HBV-related liver diseases, including HCC, however this is unclear. The goal of this study is to obtain comprehensive DNA methylation profiles of the three putative CpG islands in the HBV DNA in infected livers, with respect to liver disease progression. The extent of methylation in these CpG islands was first assessed using bisulfite PCR sequencing with a small set of tissue samples, followed by analysis using both quantitative bisulfite-specific PCR and quantitative methylation-specific PCR assays in a larger sample size (n = 116). The level of HBV CpG island 3 methylation significantly correlated with hepatocarcinogenesis. We also obtained, for the first time, evidence of rare, non-CpG methylation in CpG island 2 of the HBV genome in infected liver. Comparing methylation of the HBV genome to three known HCC-associated host genes, APC, GSTP1, and RASSF1A, we did not identify a significant correlation between these two groups.
Highlights
Despite the availability of a preventive vaccine, chronic Hepatitis B virus (HBV) infection remains a significant global health issue, affecting more than 350 million people worldwide
Only 2 studies have studied the methylation of CG3 in HBV-related HCC (HBV-HCC) tissue, but neither of them have reported an association between CG3 methylation and HCC15,16
We used genotype C for the HBV DNA sequences obtained from patient samples since this was the most prevalent strain identified in the HCC tissues examined
Summary
Despite the availability of a preventive vaccine, chronic Hepatitis B virus (HBV) infection remains a significant global health issue, affecting more than 350 million people worldwide. Previous studies have identified epigenetic modifications of HBV DNA, including methylation, in both cell cultures and HCC tissue, the association between the methylation of HBV DNA and the progression of liver disease to HCC has not yet been fully explored[15,16,17]. This could be partially due to the complexity of HBV DNA in diseased tissue[18,19,20]. Only 2 studies have studied the methylation of CG3 in HBV-HCC tissue, but neither of them have reported an association between CG3 methylation and HCC15,16
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