Abstract
目的 观察逆转录病毒介导的双自杀基因对肝内胆管癌细胞HCCC-9810的杀伤作用,探讨肝内胆管癌的基因治疗方法.方法在脂质体的介导下将含有双自杀基因的逆转录病毒载体PwzlneoCDglytk导入包装细胞PA317,经G418筛选后大量培养产病毒的阳性克隆PA317/CD+tk细胞株,收集病毒上清,转染HCCC-9810细胞,再次经G418筛选,获得稳定表达双自杀基因的HCCC-9810/CD+tk细胞株.用RT-PCR检测双自杀基因的表达.给予前体药物5-氟胞嘧啶(5-flourocytosine,5-FC)和(或)无环鸟苷(ganciciovir,GCV)后,用MTT法测定转基因组及未转基因组HCCC-9810细胞的存活率.结果双自杀基因在HCCC-9810细胞中可稳定表达,联合使用5-FC和GCV对细胞增殖的杀伤作用及旁杀伤效应高于单独使用5-FC或GCV.结论逆转录病毒介导自杀基因可有效杀死肝内胆管癌细胞,双自杀基因较单一自杀基因具有更强的抗肿瘤作用。
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