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MicroRNA‑20 induces methylation of hepatitis B virus covalently closed circular DNA in human hepatoma cells.

Molecular medicine reports (2019-07-02)
In Young Moon, Jae Hee Choi, Jung Wha Chung, Eun Sun Jang, Sook-Hyang Jeong, Jin-Wook Kim
ABSTRACT

Methylation was suggested to suppress the transcriptional activity of hepatitis B virus (HBV) covalently closed circular DNA (cccDNA) in hepatocytes. This may be associated with its low replicative activity during the inactive stage of chronic HBV infection; however, the exact mechanisms of methylation in HBV infection remain unknown. We have previously shown that short hairpin RNAs induced the methylation of the HBV genome in hepatoma cell lines. We also reported that the microRNA (miR) 17‑92 cluster negatively regulates HBV replication in human hepatoma cells. In addition, miR‑20a, a member of the miR 17‑92 cluster, has sequence homology with the short hairpin RNA that induces HBV methylation. In the present study, we investigated whether miR‑20a can function as an endogenous effector of HBV DNA methylation. The results indicated that overexpression of miR‑20a could suppress the replicative activity of HBV and increased the degree of methylation of HBV cccDNA in the HepAD38 hepatoma cell line. Argonaute (AGO)1 and AGO2, effectors of the RNA‑induced silencing complex, were detected in the nucleus of HepAD38 cells; however, only AGO2 was bound to HBV cccDNA. In addition, intranuclear AGO2 was determined to be bound with miR‑20a. In conclusion, miR‑20a may be loaded onto AGO2, prior to its translocation into the nucleus, inducing the methylation of HBV DNA in human hepatoma cells, leading to the suppression of HBV replication.

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Sigma-Aldrich
Monoclonal ANTI-FLAG® M2 antibody produced in mouse, 1 mg/mL, clone M2, affinity isolated antibody, buffered aqueous solution (50% glycerol, 10 mM sodium phosphate, and 150 mM NaCl, pH 7.4)
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Imprint® DNA Modification Kit, For bisulfite DNA conversion & purification