人肝癌细胞系中稳定表达且整合的乙型肝炎病毒基因组。

Stable expression and integrated hepatitis B virus genome in a human hepatoma cell line.

作者信息

Wang L Y, Li Y G, Chen K, Li K, Qu J L, Qin D D, Tang H

机构信息

Key Laboratory of Molecular Biology on Infectious Diseases, Ministry of Education, Second Affiliated Hospital, Chongqing Medical University, Chongqing, China.

出版信息

Genet Mol Res. 2012 May 18;11(2):1442-8. doi: 10.4238/2012.May.18.3.

DOI:10.4238/2012.May.18.3

PMID:22653591

Abstract

HepG2.2.15 cell is a widely used cell model for studying HBV (hepatitis B virus) in vitro. In these cells, the HBV genome is integrated in several sites of HepG2 cellular DNA. These multiple copies may have some influence on the cellular processes. We constructed a new plasmid, pSEH-Flag-HBV, and transfected it into HepG2 cells, and then screened it with hygromycin. We then used ELISA, PCR, and RT-PCR to detect the expression of HBV in these cell lines. A cell line that stably expressed hepatitis B e antigen (HBeAg) and hepatitis B surface antigen (HBsAg) was established. Using Southern blotting analysis, we found that the HBV genome was integrated as a single copy in the cellular DNA. This cell line will be a useful alternative model for HBV studies.

摘要

HepG2.2.15细胞是一种广泛用于体外研究乙肝病毒（HBV）的细胞模型。在这些细胞中，HBV基因组整合于HepG2细胞DNA的多个位点。这些多拷贝可能对细胞过程有一定影响。我们构建了一种新质粒pSEH-Flag-HBV，并将其转染至HepG2细胞，然后用潮霉素进行筛选。接着我们用ELISA、PCR和RT-PCR检测这些细胞系中HBV的表达。建立了一个稳定表达乙肝e抗原（HBeAg）和乙肝表面抗原（HBsAg）的细胞系。通过Southern印迹分析，我们发现HBV基因组以单拷贝形式整合于细胞DNA中。该细胞系将成为HBV研究的一个有用的替代模型。