腺病毒介导的HSV-tk/GCV系统对肺癌细胞的杀伤作用

Adenovirus vector mediated HSV-tk gene killing action on Lewis lung cancer cells

目的:用含HSV-tk重组腺病毒载体转染Lewis肺癌细胞,探讨其对肺癌细胞的杀伤作用。方法:利用同源重组技术构建含HSV-tk基因的重组腺病毒载体(AdCMVtk),将含LacZ基因的腺病毒载体(AdCMVLacZ)转染Lewis肺癌细胞,X-gal染色后,测定腺病毒对该细胞的转染率,并观察AdCMVtk/GCV系统对Lewis肺癌细胞存活率的影响。结果:随着AdCMVLacZ的病毒量(MOI)增加,对Lewis细胞转染率也增加,当MOI为500时,转染率可达100%。AdCMVtk/GCV系统可明显杀伤Lewis细胞,且随着AdCMVtk的MOI增加,或GCV浓度的增加,细胞存活率减少,但单独使用AdCMVtk或GCV时,对Lewis细胞无杀伤作用。AdCMVtk/GCV系统对该细胞的杀伤作用具有明显的旁观者效应,20%Lewis细胞被AdCMVtk转染可引起74%细胞死亡。结论:AdCMVtk/GCV系统杀伤肿瘤细胞既有效又安全。

Objective To study the killing action of adenovirus vector carrying HSV-tk （AdCMVtk） on Lewis cells. Methods The recombinant adenovirus carrying HSV-tk was constructed by homologous recombination techniques. The adenovirus transfected efficiency to Lewis cells was measured, the cells were transfected by adenovirus vector with LacZ gene, then stained with X-gal. The effect of AdCMVtk/GOV system on survival rate of Lewis cells was observed. Results The adenovirus transfected efficiency to Lewis cells was increased with the increasing of multiplicity of infection （MOI）. 100 % transfected efficiency needed MOI 500. AdCMVtk/GCV system could kill Lewis cells. When the amount of adenovirus or the concentration of GCV was increased, the cell survival rate was decreased, but AdCMVtk or GCV alone could not kill Lewis cells. The killing action existed bystander effect, 74% Lewis cells were died with only 20% Lewis cells transfected AdCMVtk. Conclusion AdCMVtk/GCV system is effective and safe on killing tumor cells.