eISSN: 1896-9151
ISSN: 1734-1922
Archives of Medical Science
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abstract:
Basic research

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

Suofeng Sun, Yuan Li, Bowei Liu, Bingyong Zhang, Shuangyin Han, Xiuling Li

Arch Med Sci
Online publish date: 2018/11/20
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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.

keywords:

hepatitis B virus, stable cell lines; hepatitis B virus genome, episomal replication; antiviral drug evaluation

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