Stable HepG2- and Huh7-based human hepatoma cell lines for efficient regulated expression of infectious hepatitis B virus
Stable HepG2- and Huh7-based human hepatoma cell lines for efficient regulated expression of infectious hepatitis B virus
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DOI：
10.1016/j.jhep.2006.05.019
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发表时间：
2006-11-01
影响因子：
25.7
通讯作者：
Nassal, Michael
中科院分区：
文献类型：
--
作者：
Sun, Dianxing;Nassal, Michael
背景/目的：乙肝病毒不能在培养细胞中繁殖，但两种人肝癌细胞系HepG2和Huh7在转染HBVDNA后支持病毒复制。如果需要标准化，稳定转染的细胞系提供了明显的优势。其中一个这样的品系，HepG2.2.15，被广泛用于抗病毒研究，但乙肝病毒的产量有限，很难控制。我们的目的是建立稳定的、可诱导产生乙肝病毒的HepG2和HuH7细胞株，以克服这些限制。方法：基于四环素(Tet)调控的TetOFF系统，将Tet反应启动子控制的HBV基因组分别导入分别建立的表达Tet反应反式激活因子(TTA)的可调控的HepG2和HuH7细胞系。对稳定的克隆进行了可调控性和表达水平、病毒产量以及对抗病毒药物的反应性分析。结果：建立了基于HepG2和HuH7的细胞系，Tet可控地比HepG2.2.15细胞产生更多的乙肝病毒。分泌的病毒粒子对原代兔肝细胞具有感染性，细胞对不同抗病毒药物的反应与HepG2.215细胞一致。结论：所建立的新的乙肝病毒细胞系可作为学术和药物研究的有价值的工具。亲本TTA细胞将促进额外品系的产生，在相同的宿主细胞背景下产生HBV变异体或其他基因。(C)2006年欧洲肝脏研究协会。爱思唯尔出版，版权所有。
Background/Aims: Hepatitis B virus (HBV) cannot be propagated in cultured cells but two human hepatoma cell lines, HepG2 and Huh7, support virus replication when transfected with HBV DNA. If standardization is required stably transfected cell lines provide distinct advantages. One such line, HepG2.2.15, is widely used in antiviral research but HBV production is limited and difficult to control. Our aim was to establish stable, inducibly HBV producing HepG2 and Huh7 cell lines that overcome these limitations.Methods: Based on the tetracycline (Tet)-regulated TetOFF system, a Tet-responsive promoter-controlled HBV genome was introduced into separately established, well-regulatable HepG2 and Huh7 lines expressing Tet-responsive trans-activators (tTAs). Stable clones were analyzed for regulatability and levels of HBV expression, quality of the virus produced, and responsiveness towards antivirals.Results: HepG2- and Huh7-based cell lines were established which, Tet-controllably, produce more HBV than HepG2.2.15 cells. The secreted virions were infectious for primary tupaia hepatocytes, and the cell lines responded as well as HepG2.215 cells to different antivirals.Conclusions: The new HBV cell lines should be valuable tools for academic and pharmaceutical HBV research. The parental tTA-cells will facilitate the generation of additional lines, producing HBV variants, or other genes, in an identical host cell background. (c) 2006 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.