摘要
目的观察单纯疮疹病毒胸苷激酶（ＨＳＶ－ｔｋ）／丙氧鸟苷（ＧＣＶ）自杀基因治疗系统在体内外对人肝癌的杀伤效应。方法构建携带ＨＳＶ－ｔｋ基因的逆转录病毒载体，将该重组逆转录病毒感染人肝癌细胞ＨＣＣ９２０４，筛选稳定表达ｔｋ的细胞克隆ＨＣＣ９２０４／ｔｋ，并测定其在体外对ＧＣＶ的敏感性；ＨＣＣ９２０４／ｔｋ细胞在裸鼠皮下成瘤，用ＧＣＶ治疗并观察疗效。结果ＧＣＶ在体外对ＨＣＣ９２０４／ｔｋ细胞有明显的杀伤作用和旁杀伤效应，裸鼠体内ｅｘｖｉｖｏ实验得到相似结果，在体内外对未转染细胞则无明显毒性．结论在ｉｎｖｉｔｒｏ水平（指完全体外）和ｅｘｖｉｖｏ水平（指一部分体外一部分体内），表达ＨＳＶ－ｔｋ基因的肿瘤细胞均可被ＧＣＶ有效杀伤，逆转录病毒介导的ＨＳＶ—ｔｋ／ＧＣＶ自杀基因治疗系统有可能成为肝癌的有效治疗方法。
Objective The efficacy of the herpers simplex virus thymidine kinase/gnnciclovir(HSV- tk/GCV) suicide-gene therapy system on human hepatocellular carcinoma was oberved in vitro as well asex vivo. Methods The recombinant retroviral vector containing HSV-tk gene was constructed andHCC9204 hutun hepatOCellular cahooma Cell line was infected with the recombinani retrovirus. FollowingSelection the HCC9204/tk Cell line which stably expressed tk was cloned. The Sensitivity of HCC9204/tkcells to GCV was examined in vitro. Antitumor effects of GCV were also oborved after the administrationof GCV in nude mice bearing tumor derived derived from HCC9204/tk Cells. Besults The HSV-tk gene expressedStably in HCC9204/tk Cells. In vitro the groWth inhibition studies showed that HCC9204/tk cells werehighly sensitive to GCV(IC50 1.2mg/L in 72 hour), and the HSV-tk gene-manied HCC9204/tk cells weretoxic to nearby unmodified HCC9204 tumor cells that were resistant to GCV. This phenomenon wastermed 'bystander effect'. Ex vivo studies showed similar results that significant tumor inhibition wasfound in the treatment group than in control group. ConcluSion Tumor cells expressing HSV-tk gene canbe eradicated by GCV in vitro and ex vivo. The HSV-tk/GCV suicide-gene therapy system mediatedretrovirally may provide an effective approach to treatment of human hepatoma.
出处
《中华肝脏病杂志》
CAS
CSCD
1999年第2期113-115,共3页
Chinese Journal of Hepatology
关键词
肝细胞癌
基因疗法
逆转录病毒
Carcinoma, hepatocellular Gene therapy Retrovirus HSV-tk
