逆转录病毒介导的人肝癌特异性HSV－tk／ACV的体外抗肝癌作用

Prodrug transformation gene therapy mediated by retroviral vector for the treatment of human hepatoma: anti-hepatoma effect of HSV-tk/ACV in vitro

目的：建立肝癌特异性前药转换基因治疗的方法。方法和结果：构建携带单纯疮疹病毒胸苷激酶基因（ＨＳＶ－ｔｋ）的人肝癌特异性逆转录病毒载体，经过ＰＡ３１７细胞包装及病毒滴度测定后，感染肝癌ＨｅｐＧ２和ＳＭＭＣ７７２１细胞，用Ｎｏｒｔｈｅｒｎ杂交方法证明ＨＳＶ－ｔｋ基因在ＡＦＰ高分泌的ＨｅｐＧ２细胞中得到特异转录，以阿昔洛韦（ＡＣＶ）为前药攻击转基因的肝癌细胞显示对ＨｅｐＧ２有相对选择性杀伤作用。结论：该ＡＦＰｅ／ＨＳＶ－ｔｋ／前药转换疗法为肝癌特异性基因治疗提供了有价值的资料。

To establish the method for hepatoma- specific prodrug transformation gene therapy. Methods and Results: Construction of retroviral vector carrying HSV-tk gene flanked by human α-fetoprotein(AFP) enhancer core sequence and human pgk promoter; Replication-defective vector viral particles were obtained by transfer of the vector DNA into the packaging cell line PA317, and used to infect human hepatoma cell line HepG2 and SMMC7721. Northern blot analysis showed that HSV-tk gene was specifically transcribed in HepG2 cell in which α- fetoprotein was highly produced. The sensitivity of both modified hepatoma cells to the ACV was found to be significant more than their parental cells. The increasing extent of HepG2 cells was more significant than SMMC7721 cells. Conclusion: AFPe/HSV-tk/ACV therapy can provide available materials for the hepatoma-specific prodrug transformation gene therapy.