Methylation Pattern of Hepatitis B Virus X Gene in Infected Patients

Authors
1 Department of Virology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran
2 Department of Medical Genetic, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran
Abstract
Objective: A recent field of research in epigenetics is DNA methylation which involves the CpG island in the genome that subsequently controls transcription and translation of targeted genes. In the hepatitis B virus (HBV) genome, there are three CpG islands which tend to be methylated. The aim of the current study is to determine the methylation pattern of the HBV X gene in chronically infected HBV patients.
Methods: Study participants comprised 45 chronically infected HBV patients. According to the presence of the HBeAg, patients were divided into two groups, HBeAg positive (n=24) and HBeAg negative (n=21). Initially, viral DNA was treated with natrium bisulfate. Then, analysis was performed with two sets of methylated and non-methylated primers by the MSP method.
Results: The overall methylation rate in serum samples of hepatitis B infected patients was 35.5%; the rate in the HBeAg positive patients was 20.8%, whereas it was 52.3% in HBeAg negative patients. There was a significantly higher rate of methylation in serum samples of HBeAg negative patients compared to HBeAg positive patients (student's t-test; P=0.02).
Conclusion: Methylation of HBV can be used as a new mechanism to control the progression of viral infection. This methodology can be useful for determining the characteristics of clinical stages of this infection.

Keywords


[1]     Pan CQ, Zhang JX. Natural History and Clinical Consequences of Hepatitis B Virus Infection. Int J Med Sci 2005; 2(1): 36-40.
[2]     Kaur P, Paliwal A, Durantel D, Hainaut P, Scoazec JY, Zoulim F, Chemin I, Herceg Z. DNA methylation of hepatitis B virus (HBV) genome associated with the development of hepatocellular carcinoma and occult HBV infection. J Infect Dis 2010; 202(5): 700-4.
[3]     Vivekanandan P, Thomas D, Torbenson M. Hepatitis B viral DNA is methylated in liver tissues. J Viral Hepat 2008; 15(2): 103-7.
[4]     Guo Y, Li Y, Mu S, Zhang J, Yan Z. Evidence that methylation of hepatitis B virus covalently closed circular DNA in liver tissues of patients with chronic hepatitis B modulates HBV replication. J Med Virol 2009; 81(7): 1177-83.
[5]     Hoelzer K, Shackelton LA, Parrish CR. Presence and role of cytosine methylation in DNA viruses of animals. Nucleic Acids Res 2008; 36(9): 2825-37.
[6]     Li HP, Leu YW, Chang YS. Epigenetic changes in virus-associated human cancers. Cell Res 2005; 15(4): 262-71.
[7]     Sutter D, Doerfler W. Methylation of integrated adenovirus type 12 DNA sequences in transformed cells is inversely correlated with viral gene expression. Proc Natl Acad Sci U S A 1980; 77(1): 253-6.
[9]     Chen JY, Hsu HC, Lee CS, Chen DS, Zuckerman AJ, Harrison TJ. Detection of hepatitis B virus DNA in hepatocellular carcinoma: methylation of integrated viral DNA. J Virol Methods 1988; 19(3-4): 257-63.
[10]  Araki K, Miyazaki J, Tsurimoto T, Inomoto T, Iwanaga T, Matsubara K, Yamamura K. Demethylation by 5-azacytidine results in the expression of hepatitis B virus surface antigen in transgenic mice. Jpn J Cancer Res 1989; 80(4): 295-8.
[11]  Vivekanandan P, Thomas D, Torbenson M. Methylation regulates hepatitis B viral protein expression. J Infect Dis 2009; 199(9): 1286-91.
[12]  Vivekanandan P, Kannangai R, Ray SC, Thomas DL, Torbenson M. Comprehensive genetic and epigenetic analysis of occult hepatitis B from liver tissue samples. Clin Infect Dis 2008; 46(8): 1227-36.