腺病毒介导的HSV-tk体外抗喉癌作用

Anti-laryngeal carcinoma with adenovirus vector mediated HSV-tk gene

目的观察 HSV- tk/ GCV系统对喉癌细胞的杀伤作用。方法将带有报告基因 L ac Z的 5型缺陷性腺病毒载体Ad CMVL ac Z感染喉癌细胞 Hep- 2 ,X- gal染色 ,测定腺病毒的转染率。利用同源重组技术构建含 HSV- tk基因的重组腺病毒载体 Ad CMVtk。Ad CMVtk感染 Hep- 2细胞后 ,加入 10 mg/ L GCV,观察瘤细胞存活率及旁观者效应。结果随着腺病毒感染复数量 (multiplicity of infection,MOI)增加 ,对 Hep- 2细胞转染率亦增加 ,10 0 %转染所需的 MOI为 2 0 0。 Ad CMVtk/ GCV对Hep- 2细胞的杀伤强度与 Ad CMVtk量呈正向关系 ,即有浓度依赖性。此杀伤作用存在旁观者效应 ,但较弱。结论腺病毒介导的 HSV- tk/ GCV具有杀伤喉癌细胞作用。

ObjectiveTo observe killing action of HSV-tk/GCV system on laryngeal cancer cells. Methods Hep-2 cells were transfected by adenovirus vector with LacZ gene, then stained with X-gal for measurement of the adenovirus transfected efficiency. The recombinant adenovirus carrying HSV-tk was constructed by homologous recombination techniques. Hep-2 cells survival and by stander effect were observed after adding 10 mg/L GCV to transfected cells. Results The more the multiplicity of infection (MOI), the more the adenovirus transfected efficiency to Hep-2 cells. 100% transfected efficiency needs MOI 200 The killing action of HSV-tk/GCV on Hep-2 cells depends on the amount of AdCMVtk. There is a weak by stander effect. Conclusion Adenovirus mediated HSV-tk/GCV can kill Hep-2 cells. [