腺病毒介导的HSV-tk体外抗喉癌作用

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 Lac 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/ LGCV,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

Anti-laryngeal carcinoma of adenovirus vector-mediated HSV-tk gene in vitro

Objective:To 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 measuring the adenovirus transfected efficiency.The recombinant adenovirus carring HSV tk was constructed by homologous recombination techniques.Hep 2 cells survival and bystanding 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 needed MOI 200. The killing action of HSV tk/GCV on Hep 2 cells.Depended on the amount of AdCMVtk.There was weak bystanding effect.Conclusion:Adenovirus mediated HSV tk/GCV can kill Hep 2 cells.