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The Effect of NIRF on the Acetylation Status of HBV cccDNA Bound-H3 in HepG2 Cells
Zhou Danlin1, Hu Bin1, Xuan Yanyan1, Duan Changzhu1,2*
1Department of Cell Biology and Medical Genetics, Chongqing Medical University, Chongqing 400016, China; 2Key Laboratory of Clinical Laboratory Diagnostics of Ministry of Education, Chongqing Medical University, Chongqing 400016, Chi
Abstract: To determine the effect of NIRF protein (Np95/ICBP-90 like RING finger protein) on the replication of HBV and the acetylation of cccDNA-bound H3 histone at various times after transient transfection of linear HBV DNA into human hepatoma HepG2 cells. HepG2 cells were infected with GEM-HBV1.3+pFLAG、pGEM-HBV1.3+pFLAG-NIRF、pGEM-HBV1.3 plasmids. The secretion of HBsAg and HBeAg in the cultural supernatants of the transfected cells was detected by ELISA. The expression of HBV mRNA and NIRF protein was examined by RT-PCR and Western blot， respectively. Finally the levels of the HBV cccDNA-bound H3 histone and the acetylated H3 histone were identified by ChIP (Chromatin Immunoprecipitation). Our results demonstrated that the acetylation of cccDNA-bound H3 histone was associated with the level of HBV replication. NIRF can inhibit the levels of cccDNA-bound H3 and acetylated H3 in HepG2 cells， which may play roles in a better understanding of the mechanisms of HBV and confirm new therapeutic strategies against hepatitis B virus.
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