Establishment of stable cell lines in which the HBV genome replicates episomally for evaluation of antivirals

建立HBV基因组以游离体形式复制的稳定细胞系,用于评估抗病毒药物

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Abstract

INTRODUCTION: Due to the increasing resistance to nucleot(s)ide analogs in patients with chronic hepatitis B, development of new antiviral drugs to eradicate hepatitis B virus is still urgently needed. MATERIAL AND METHODS: To date, most studies on evaluating anti-HBV drugs have been performed using cell lines where the HBV genomic DNA is chromosomally integrated, e.g. Hep2.2.15 in HBV-infected livers of the viral episomal genome replicates in the nucleus and covalently closed circular DNA (cccDNA) serves as a transcriptional template. Another option involves the use of HBV-infected cells of HepaRG or NTCP-overexpressing cells. However, the development of the infection system is expensive and laborious, and its HBV expression level remained low. RESULTS: Compared to HuH7 cells, the established stable cell lines based on episomal-type pEB-Multi vectors can been expressed HBV wild-type by qRT-PCR and immunoblotting (p < 0.05). These two vectors are also sensitive to Entecavir and against nucleoside analog Lamivudine in mutants cellines. CONCLUSIONS: It is worth demonstrating how useful the established cell system is for evaluating antiviral agents and their mechanisms of action.

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