腺病毒 HSV1

孔北华　姜洁　刘春生　江森

　　【摘要】　目的：研究腺病毒/HSV1-tk／GCV、ACV系统对卵巢上皮癌细胞旁观者效应。方法：常规重组腺病毒扩增纯化后，测病毒滴度。X-gal染色法测定转染效率，PCR检测HSV1-tk的表达，混合细胞培养实验观察旁观者效应。结果：重组腺病毒扩增纯化后测定滴度分别为Ad/tk?6.2×109pfu/ml，AdLacZ?1.4×109pfu/ml。当感染复数为100时，近100%的细胞可被感染。PCR结果显示，Adtk转染TYK后，可见404bp的特异性条带，在同一MOI中，有10%的Adtk/TYK细胞存在，GCV就可杀死近50%左右的混合细胞。ACV的作用约为GCV的1/10。结论：重组腺病毒Ad/tk可有效地感染人卵巢上皮癌细胞，腺病毒/HSV1-tk／GCV、ACV系统存在着旁观者效应。
　　【关键词】　卵巢上皮癌细胞；旁观者效应；腺病毒；基因，HSV1-tk；
　　中图分类号　R737.31

Study on the bystander effect of the Ad/HSV1-tk/GCV，ACV system on ovarian epithelial cancer cells.

Kong Beihua,Jiang Jie
（Dept.of Gynecology and Obstetrics Shandong Medical University,Jinan，250012）
Liu Chunsheng
（Cancer Center,the Affiliated Hospital of Shandong Medical University,Jinan，250012）

　　Abstract Objective：To study on the bystander effect of Ad/HSV1-tk/GCV，ACV gene therapy system on ovarian epithelial cancer cells.Methods:Recombinant adenovirus was amplificated and purified using routine method,the titer was detected.The efficiency of recombinant Adtk transference were evaluated by regular X-gal staining.The expression of HSV1-tk gene was assayed using PCR.The bystander effect was observed by mixed cell culture.Results:Recombinant adenovirus was amplificated and purified.The titer of Ad/tk was 6.2×109pfu/ml and of AdLacZ was 1.4×109pfu/ml respectively.When the MOI was 100,approximately 100% cells were infected.PCR assay showed specific 404bp band.In the same MOI,GCV could killed almost 50% cells when there were 10% Ad/tk＋.The effect of ACV was about 10 times weaker than that of GCV.Conclusions:Replicated-deficient recombinant adenovirus Adtk can affected human ovarian epithelial cancer cells effectively.Bystander effect is found in Ad/HSV1-tk/GCV，ACV gene therapy system.
　　Key words　Ovarian epithelial carcinoma cell line;Bystander effect;Adenovirus;Gene，HSV1-tk
 
　　将Ⅰ型单纯疱疹病毒胸苷激酶基因(HSV1-tk)转染入肿瘤细胞后，基因的表达产物可将本来无毒的核苷类似物羟甲基无环鸟

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