【摘要】 目的:探讨反转录病毒介导的肿瘤坏死因子α(TNF-α)基因对C6胶质瘤的抗瘤效应,从而为肿瘤的基因治疗提供基础研究数据。方法:将重组TNF-α反转录病毒载体PLJ+TNF转染大鼠C6胶质瘤细胞,ELISA法检测该细胞中目的基因的表达,MTT法检测体外增殖能力;将实验大鼠随机分为实验组和对照组,每组10只,实验组大鼠右股内侧接种转基因2×106C6细胞,对照组同法同部位接种未转基因C6细胞,常规饲养5周,每周测量肿瘤大小,绘制肿瘤生长曲线,比较两组皮下肿瘤大小并进行统计学分析。结果:C6胶质瘤细胞与C6细胞的体外增殖能力比较,差异无显著性意义(P>0.05);转基因前后TNF-α分泌情况:1×106个C6细胞体外培养24h每毫升上清液分泌TNF-α(2.42±0.76)U;1×106个C6胶质瘤细胞体外培养24h每毫升上清液分泌TNF-α(17.53±2.31)U,两者比较,差异有显著性意义(P<0.01);动态TNF-α检测显示:转基因C6细胞能稳定地表达高水平的TNF-α;大鼠皮下接种肿瘤细胞5周后,实验组3只未见肿瘤生长,其余肿瘤直径为(1.5±0.3)cm,对照组肿瘤直径为(2.4±0.2)cm,两组比较,差异有显著性意义(P<0.01)。结论:TNF-α基因转染大鼠C6胶质瘤细胞后能持续分泌高水平的TNF-α,对大鼠C6胶质瘤皮下肿瘤具有明显的抗瘤效应。更多还原

【Abstract】 AIM:To study the effect of tumor necrosis factor alpha (TNF α ) induced by retrovirus against C6 glioma,so as to provide data for basic investigation of cancer gene therapy.METHODS:C6 gliomas of rats were transfected with viral vector PLJ TNF of TNF α .The expression of genes was detected with enzyme of linked immunoadsordent assay(ELISA),and the ability of proliferation in vitro was estimated with MTT reduction assay.Totally 20 rats were randomly divide into experimental group and control group, 10 rats in each group.The transgenic 2× 106 C6 cells were inoculated into the inside of right thigh of rats in experimental group.Nontransgenic 2× 106 C6 cells were inoculated in control group.All rats were fed for five weeks normally.The size of tumor was detected every week,and the growth curve of tumor was drawn.The size of tumor was compared and analyzed. RESULTS:There was no significant difference in the ability of proliferation in vitro between C6 glioma cells and C6 cells(P >0.05).The excretion of TNF α before and after transgene: supernatant of each milliliter of 1× 106 C6 cells cultured in vitro for 24 hours excreted TNF α of (2.42± 0.76) U,supernatant of each milliliter of 1× 106 C6 glioma cells cultured in vitro for 24 hours excreted TNF α of (17.53± 2.31) U,and there was significant difference(P< 0.01).The results of dynamic estimation of TNF α indicated:transgene C6 cells expressed TNF α with high levels steadily.Five weeks after endermic inoculation of tumor cells of rats,the tumor size of three rats in experimental group did not change and the diameter of tumor of others was(1.5± 0.3) cm.The diameter of tumor in control group was(2.4± 0.2)cm,and there was significant difference between the two groups(P< 0.01).CONCLUSION:C6 glioma cells transfected with TNF α gene can excrete TNF α with high level, and has obvious effect against dermic tumors of C6 glioma.更多还原