丙型肝炎病毒体外感染肝癌细胞株HepG2的初步研究  

A preliminary study on hepatitis C virus infection of a human liver cell line HepG2 in vitro

作　　者：闵峰[1] 王素美[1] 郝飞[2] 王宇明[2] 刘冰 

机构地区：[1]解放军第174医院传染科,厦门361003 [2]第三军医大学西南医院感染科 [3]解放军第520医院传染科

出　　处：《中华传染病杂志》2002年第6期330-332,共3页Chinese Journal of Infectious Diseases

基　　金：国家自然科学基金资助项目(396 70 6 72 )

摘　　要：目的　研究人肝癌细胞株HepG2对丙型肝炎病毒 (HCV)的易感性 ,建立细胞感染模型。方法　将人肝癌细胞株HepG2与慢性丙型肝炎患者血清共同温育 6～ 8h ,收集不同时相点标本 ,用逆转录聚合酶链反应 (RT PCR)法分别检测细胞内或上清液中正负链RNA ,免疫组织化学法检测细胞内HCVNS3、NS5特异性抗原的表达情况 ,以及原位杂交法检测细胞内HCV负链RNA。结果　接种感染血清 3～ 3 5d ,可在细胞内或培养上清液中检出HCV正负链RNA ;HCVNS3、NS5特异性抗原能在感染细胞内稳定表达 ,阳性物质位于胞浆中 ;原位杂交法证实细胞内存在负链RNA ,也位于胞浆中。结论　人肝癌细胞株HepG2不但对HCV易感 。Objective To establish practical cell model of HCV infection, and investigate the susceptibility of a human liver cell line HepG2 to hepatitis C virus in vitro. Methods A human liver cell line HepG2 was incubated with serum from a chronic hepatitis C patient for 6～8 hours. Both the plus and minus strands of HCV RNA in infected cells or supernatant were examined by reverse transcription polymerase chain reaction (RT PCR). The HCV NS 3,NS 5 antigens in infected cells were respectively detected with the monoclonal antibodies to antigens of their own by immunohistochemical assay. The minus strand of HCV RNA in infected cells were localized by in situ hybridization. Results The intracellular plus or minus stands of HCV RNA were first detected on day 3 post incubation and then could be intermittently detected until day 35 post incubation in cells or supernatant. The positive signals of NS 3,NS 5 antigens could be expressed within cytoplasm of infected cells. The minus strand of HCV RNA was located within cytoplasm by in situ hybridization. Conclustions These results show that a human liver cell line HepG2 is not only susceptible to HCV but also able to support its long time replication in vitro.

关 键 词：丙型肝炎病毒 体外感染 肝癌 HEPG2细胞 RT-PCR 

分 类 号：R735.7[医药卫生—肿瘤]