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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HCVRNA阳性血清感染人肝癌细胞株HepG2 ,培养6 0d ,用套式逆转录聚合酶链反应(nestedRT PCR)检测培养细胞及上清中正、负链HCVRNA。结果 HepG2 在感染后第2～30天,细胞中可以间断检出HCV正链RNA ;负链RNA在感染后第3～30天可以间断检出,检出率与正链RNA接近。在第31～6 0天仍可间断检出HCVRNA正、负链,但检出率逐渐降低。HepG2 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。培养上清中均未检出HCV负链RNA。结论 HepG2 细胞对HCV易感,并能支持HCV长期复制,HepG2 细胞是较为理想的HCV体外感染细胞模型。
Objective To establish a model of Hepatitis C virus (HCV) infection that is close to the natural infection in vivo and can be replicated for a long time. Methods Human hepatocellular carcinoma cell line HepG2 was infected with HCV RNA positive serum and cultured for 60 days. The positive and negative HCV RNA in cultured cells and supernatant were detected by nested RT PCR. Results HepG2 could detect HCV positive stranded RNA intermittently on the 2nd to 30th day after infection. Negative stranded RNA could be detected intermittently on the 3rd to 30th day after infection, and the detection rate was close to the positive stranded RNA. On the 31st to 60 days, positive and negative HCVRNA could still be detected intermittently, but the detection rate decreased gradually. HepG2 culture supernatant HCV positive strand RNA infection also showed intermittent positive, the detection rate and intracellular positive strand RNA are basically the same. None of the HCV negative strand RNA was detected in the culture supernatant. Conclusion HepG2 cells are susceptible to HCV and can support long-term HCV replication. HepG2 cells are an ideal model of HCV in vitro infection.