目的 研究重组并纯化的肿瘤坏死因子凋亡诱导配体蛋白(TRAIL)对前列腺癌(PC-3)及膀胱癌细胞(5637)的抑制作用.方法 PC-3 和5637 细胞分为对照组(仅加培养液)和实验组(加入40 ng/ml 的TRAIL 蛋白处理),细胞培养12 h 后加入TRAIL 蛋白,24 h 后应用PCR、Western blot 技术检测细胞内抗凋亡蛋白Bcl-2 及凋亡信号通路中Caspase-3 和Caspase-8 基因和蛋白的表达.结果 应用SPSS 统计分析软件,组间比较采用t 检验,结果显示PC3 和5637 细胞在TRAIL蛋白作用24 h 后Bcl-2 基因和蛋白表达水平均降低,而Caspase-8、Caspase-3 基因和蛋白表达则增加.与对照组相比,其差异具显著性意义.结论 TRAIL 蛋白通过下调抗凋亡基因Bcl-2 的表达,增强Caspase-8、Caspase-3 的表达从而诱导两种肿瘤细胞的凋亡达到抑癌效应.
ObjectiveThe purpose of this study is to observe the inhibitory action of recombinant and purified TRAIL in the prostate cancer (PC3) and bladder cancer cell (5637).MethodsThe PC3 and 5637 cell lines were divided into control group (only with medium) and experimental group (with 40 ng/ml TRAIL). The TRAIL protein was added in medium after the cells were cultured for 12 hin vitro. For 24 h, real-time PCR and Western blot analysis were used to investigate the genes and protein expression of Bcl-2, Caspase-3 and Caspase-8.Results All data are analyzed by two-tailed Student’st-test using SPSS software. Results revealed that the expression of Bcl-2 decreased while the expression of Caspase-8, Caspase-3 increased in PC3 and 5637 cell lines with TRAIL for 24 h. Compared with the control group, the results had significant differences.ConclusionRecombinant and purified TRAIL induced apoptosis of two kinds of tumor cells to reach the antitumor effect through down-regulating the expression of anti-apoptosis gene Bcl-2 and increasing the expression of apoptosis genes Caspase-8, Caspase-3.