Abstract
目的 观察腺病毒介导的胞嘧啶脱氨酶/单纯疱疹病毒-1-胸苷激酶(CD/TK)双自杀基因对人胆管癌细胞QBC939的体外杀伤作用.方法CD/TK克隆入穿梭载体成为pAdtrack-CMV-CD/TK,与骨架载体pAdeasy-1在细菌内同源重组为pAd-CD/TK,经Pac Ⅰ酶切,293细胞包装、扩增、纯化后,体外转染人胆管癌细胞QBC939,并给予前药5-FC或GCV,观察其体外杀伤效果.结果含CD/TK基因的重组腺病毒鉴定正确,扩增纯化后,病毒滴度为1×1011颗粒/ml.重组腺病毒对QBC939细胞在感染倍数(m.o.i)为100时的转染效率为90%,在m.o.i50感染时,0.1 mmol/L的5-FC及10μmol/L的GCV对QBC939细胞的杀伤率为80%,明显高于单用5-FC与GCV的效应.结论双自杀基因以腺病毒为载体对人胆管癌细胞转染效率高,体外杀伤效应明显.腺病毒介导的双自杀基因治疗有望成为治疗胆管癌的有效方法。
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