丙型肝炎病毒感染细胞模型的实验研究

目的 建立接近体内自然感染状态并能长期复制的丙型肝炎病毒(HCV)感染细胞模型．方法 用HCV阳性血清感染人肝癌细胞株HepG2、SMMC 7721及胎肝细胞株L02，继续培养60d，用巢式逆转录-聚合酶链反应(nested RT-PCR)检测培养细胞及上清中正、负链HCV RNA。结果 HepG2、SMMC 772l在感染后第2-30d，L02在感染后第3-30d细胞中可以间断检出HCV正链RNA；3株细胞的细胞内HCV负链RNA均在感染后第3-30d可以间断检出，检出率与正链RNA接近。3株细胞以HepG2细胞中HCV RNA正、负链检出率较高，并在第3l—60d仍可间断检出，但复制程度逐渐减弱。3株细胞的培养上清中HCV正链RNA感染后也呈间断阳性，检出率与细胞内正链RNA基本一致。培养上清中均末检出HCV负链RNA。结论 HePG2、SMMC772l及L02细胞均对HCV易感，并能支持HCV长期复制。而HePG2细胞是较为理想的HCV体外感染细胞模型。

Establishment of Hepatitis C Virus Infected Cell Model

Objective To establish a hepatitis C virus (HCV) infected cell model which is similar to the infection in vivo and can support HCV to replicate for a long time. Methods After infected by HCV-positive serum, HepG2, SMMC 7721 and L02 cells were cultured for 60 days. Nested RT-PCR was used to detect plus and minus HCV RNA in cultured cells and supernatant. Results Plus HCV RNA was detected disconnectedly in HepG2 and SMMC 7721 cells during 2-30 days, and in L02 cells during 3-30 days after infection. Minus HCV RNA was detected in all of three cell lines during 3-30 days after infection, whose detectable rate was similar to plus HCV RNA. The detectable rate of HCV RNA was highest in HepG2, and could be still disconnectedly detected during 31-60 days after infection. However, the detectable rate was gradually reduced. Plus HCV RNA was also disconnectedly positive in three cell lines in the supernatant, and the detectable rate was consistent with that in cells. Minus HCV RNA was not detected in the supernatant. Conclusion HepG2, SMMC 7721 and L02 were susceptible to HCV, and could support HCV to replicate for a long time. HepG2 is the most ideal HCV infected cell model among the three cell lines.