【摘要】 目的研究p53基因优化恶性胶质瘤自杀基因治疗效果的可行性。方法以腺病毒载体转导p53和HSV-TK入C6鼠胶质瘤细胞,给以不同浓度的GCV,MTT法监测细胞生存率,TUNEL法检测凋亡。结果p53基因明显提高胶质瘤细胞对HSV-TK/GCV的敏感性,增加细胞凋亡。结论p53和HSV-TK的联合基因治疗有望成为一种较佳的优化自杀基因治疗方案。更多还原

【Abstract】 Objective To study the feasibility of optimizing curative effect of suicide gene(HSV-tk-GCV)on malignant gliomas by wt-p53.Methods AdCMV-p53was transfected into C6glioma cells at MOI of(Multiplicity of infection)0(G100),10(TPG1),and100(TPG2),then AdCMV-tk was transducted to C6glioma cells of G100,TPG1and TPG2respectively at MOI of100,and the C6glioma cells transducted with both AdCMV-p53and AdCMV-tk were exposed to various concentration of GCV.The cell survival rate was measured by MTT assay.Cell apoptosis was evaluated by TUNEL method.Expression of HSV-tk gene was identified by in situ hybridization.Expression of exogenous p53gene was identified by Western blot.Results The antitumor effect of HSV-tk/GCV was significantly enhanced by wt-p53.The concentration of GCV for ID 50 of TPG2cells(0.001μg /ml GCV)was significantly lower than that for the cells of tk-GCV0.01μg /ml group(MOI=100),while the concentration of GCV for ID 100 of TPG2(0.01μg /ml GCV)and TPG1(0.1μg /ml GCV)was greatly lower than that for the cells of tk-GCV(1μg /ml)group(MOI=100).Apoptosis of C6glioma cells were also increased by transfecting with wt-p53gene.Conclusion The combination of HSV-tk/GCV system with p53transduction may be relatively optimal scheme of suicide gene therapy for malignant gliomas.[更多还原