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目的 探讨ＨＳＶ－ｔｋ基因对各种肿瘤细胞的杀伤作用及人、鼠瘤细胞对其敏感性及瘤细胞表面的αｖ 整合素对腺病毒载体转染的影响。方法用同源重组法构建 ＡｄＣＭＶ ｔｋ基因；蚀斑形成法对重组腺病毒进行扩增、 滴定；Ｘ－ｇａｌ染色法观察腺病毒转染率，结晶紫染色法检测 ＡｄＣＭＶｔｋ／ＧＣＶ的杀伤作用。结果 ＡｄＣＭＶｔｋ对肿瘤细胞 的杀伤强度在一定范围内与ＡｄＣＭＶｔｋ的剂量呈正相关；对人类的Ｈｅｌａ、ＧＲＣ、Ｈｅｐ－２瘤细胞达１００％转染所需的腺病 毒感染复数量（ｍｕｌｔｉｐｌｉｃｉｔｙ ｏｆ ｉｎｆｅｃｔｉｏｎ，ｍ．ｏ．ｉ）较鼠Ｌｅｗｉｓ、ＢＴＴ７３９瘤细胞的ＭＯＩ量低；在相同ＭＯＩ病毒量时，有αｖ整合 素的瘤细胞转染率高于无αｖ整合素的瘤细胞。结论ＡｄＣＭＶｔｋ／ＧＣＶ具有较明显的杀伤肿瘤细胞的作用，且有种属 差异，对人瘤细胞作用大于鼠瘤细胞，瘤细胞表面的αｖ整合素有利于腺病毒的转染。
Objective To investigate the killing effect of HSV-tk gene on various tumor cells and its sensitivity to human and mouse tumor cells and the effects of αv integrin on the transfection of adenovirus vector. Methods AdCMV tk gene was constructed by homologous recombination method. The recombinant adenovirus was amplified and titrated by plaque forming method. The transfection efficiency of adenovirus was observed by X-gal staining and the killing effect of AdCMVtk / GCV by crystal violet staining. Results The cytotoxicity of AdCMVtk to tumor cells was positively correlated with the dose of AdCMVtk within a certain range. The multiplicity of infection (mUm) for human Hela, GRC and Hep-2 tumor cells was 100% .o.i) The MOI of BTT739 tumor cells was lower than that of Lewis Lewis mice. The transfection rate of tumor cells with αv integrin was higher than that of αv integrin-free tumor cells at the same MOI viral load. Conclusion AdCMVtk / GCV has the obvious effect of killing tumor cells, and there are species differences. The effect of AdCMVtk / GCV on human tumor cells is greater than that of rat tumor cells. The αv integrin on the surface of tumor cells is favorable for adenovirus transfection.