Ad-1.2乙型肝炎病毒感染HepG2细胞后HBV cccDNA的动态分析

Dynamic analysis of HBV cccDNA in HepG2 cells infected with Ad-1.2 HBV

目的研究Ad-1.2HBV感染HepG2细胞后HBV的复制情况。方法携带1.2拷贝HBVDNA的腺病毒体外感染人肝癌细胞株HepG2,ELISA法检测培养上清中HBsAg、HBeAg的动态表达;利用质粒抽提试剂盒提取细胞内cccDNA,经绿豆核酸酶处理后,用特异引物进行实时定量PCR检测。结果Ad-1.2HBV感染HepG2细胞后,可有效表达HBsAg及HBeAg;感染后第1天即可在细胞内检出cccDNA,第4天达到高峰。结论Ad-1.2HBV感染细胞,可作为抗病毒药物筛选和评价的模型。

Objective To study the replication of hepatitis B virus （HBV） in HepG2 cells infected with Ad-1.2 HBV. Methods HepG2 cells were transfected with adenovirus containing 1.2 copies of HBV DNA. The expression of HBV antigens were detected in the culture medium by means of enzyme-linked immunosorbent assay （ELISA）, and the covalently closed circular DNA （cccDNA） in the cells was extracted with plasmid extraction kit and detected by real-time PCR with selective primer after treatment with mung bean nuclease. Results I-IBsAg, HBeAg and HBV cccDNA were all detected in HepG2 cells after tranfection with Ad-1.2 HBV. HBV cccDNA was detected 1 day after the infection, reaching the peak level 4 days after infection. Conclusion Ad-1.2 HBV-infected cells can serve as the model for screening and evaluation of antiviral agents.