【摘要】 目的:观察聚乙二醇-聚谷氨酸(PEG-PBLG)纳米微球介导双融合自杀基因CDglyTK对舌鳞癌Tca8113细胞的杀伤作用,为口腔鳞癌的治疗探索新的途径和方法。方法:以PEG-PBLG纳米微球为载体介导携双自杀基因CDglyTK的真核表达质粒pcDNA3.1(+)-CDglyTK转染Tca8113细胞,通过RT-PCR检测CDglyTK的mRNA表达,MTT法描绘细胞生长曲线,流式细胞仪检测细胞凋亡并评价其疗效。统计分析采用SPSS10.0统计软件包完成,两样本均数的比较采用t检验。结果:PEG-PBLG纳米微球可介导质粒pcDNA3.1(+)-CDglyTK转染Tca8113细胞,RT-PCR可检测到转染细胞中CDglyTK的mRNA表达。转染后,细胞在滴加5-FC和GCV后生长受到明显抑制,5-FC和GCV联合使用对Tca8113细胞的生长抑制作用明显优于单用5-FC或GCV。5-FC和GCV联合使用组,凋亡指数显著高于对照组(P<0.01),同时也高于单独使用5-FC或GCV组(P<0.05)。结论:PEG-PBLG纳米载体介导双自杀基因CDglyTK,可有效杀伤舌鳞癌Tca8113细胞,为舌鳞癌的基因治疗提供了新的途径。更多还原

【Abstract】 PURPOSE: The objective of this study was to observe the therapeutic effect of CDglyTK gene mediated by PEG-PBLG nanoparticles in the treatment of tongue squamous cell carcinoma in vitro. METHODS: CDglyTK gene in pCEA-CDglyTK was subcloned into pcDNA3.1(+) to construct eukaryotic cell expression plasmid pcDNA3.1(+)-CDglyTK, and then the recombinant plasmids were transfected into Tca8113 cells by PEG-PBLG nanoparticles. The expression of CDglyTK gene was detected by RT-PCR. Cytotoxicity was evaluated by MTT and the apoptosis was examined by flow cytometry (FCM). The data were analysed statistically with SPSS10.0 package.Student t test was used for comparing the differences. RESULTS: The eukaryotic cell expression plasmid pcDNA3.1(+)-CDglyTK was constructed successfully. CDglyTK mRNA was found in the transfected Tca8113 cells by RT-PCR. The viability of transfected Tca8113 cells with 5-FC and GCV was significantly lower than that of transfected Tca8113 cells with 5-FC or GCV alone. The apoptosis index (AI) of transfected Tca8113 cells with 5-FC and GCV was significantly higher than that of untransfected Tca8113 cells(P<0.01),at the same time, higher than that of transfected Tca8113 cells with 5-FC or GCV alone(P<0.05). CONCLUSION: The results suggest that suicide gene therapy with PEG-PBLG mediating CDglyTK proved strong cytotoxicity to Tca8113 cells in vitro. It will be possible to validate this strategy in the treatment of tongue squamous cell carcinoma. Supported by National Natural Science Foundation of China (Grant No.30271423) and Natural Science Foundation of Guangdong Province (Grant No.21865).更多还原