论文部分内容阅读
目的探讨TRAIL对胶质瘤细胞凋亡的影响,并进一步研究DNA破坏药物顺铂(CDDP)在与其联合作用诱导细胞凋亡中影响作用的机制。方法行人脑胶质瘤U251细胞株的培养,应用不同浓度TRAIL和CDDP作用于胶质瘤U251细胞,MTT法检测TRAIL和CDDP分别及联合作用于U251细胞后的存活率,并以亚毒剂量的TRAIL和CDDP联合作用后,流式细胞仪检测其凋亡率。Westernblot方法检测CDDP作用前后TRAIL受体DR5表达量的变化。结果MTT结果:在一定时间范围内,随着药物浓度增加,TRAIL和CDDP分别及联合作用皆使胶质瘤U251细胞存活率逐渐降低,其中两者的联合作用效果明显,与分别作用组差异有统计学意义(P<0.01)。亚毒剂量的TRAIL(50ng·ml-1)、CDDP(4μg·ml-1)分别及联合作用人脑胶质瘤U251细胞24h后,存活率分别为:72.43%,75.56%,23.45%。亚毒剂量的TRAIL(50ng·ml-1)、CDDP(4μg·ml-1)单独或联合作用组致胶质瘤细胞凋亡率分别为25.61%、20.59%、62.33%。单独应用组与联合组之间差异有统计学意义(P<0.01)。Westernblot法检测显示:CDDP作用后较作用前DR5表达有明显增高,随CDDP剂量增多,DR5表达量逐渐增加。结论TRAIL和CDDP联合作用能显著增强其对人U251胶质瘤细胞凋亡的诱导,其机制可能与CDDP能上调TRAIL死亡受体DR5的表达有关。
Objective To investigate the effect of TRAIL on the apoptosis of glioma cells and further investigate the mechanism of DNA damage drug cisplatin (CDDP) in its combined effects on apoptosis. Methods The human glioma U251 cells were cultured and treated with different concentrations of TRAIL and CDDP in glioma U251 cells. The survival rates of TRAIL and CDDP were detected respectively by MTT assay and U251 cells. After the combination of TRAIL and CDDP, the apoptosis rate was detected by flow cytometry. Western blot was used to detect the expression of TRAIL receptor DR5 before and after CDDP treatment. Results MTT results: With the increase of drug concentration, the survival rate of glioma U251 cells decreased with the increase of drug concentration and TRAIL and CDDP, respectively, and the combination effect of them was obvious. There was significant difference between the two groups Statistical significance (P <0.01). The survival rates of sub-lethal doses of TRAIL (50ng · ml-1), CDDP (4μg · ml-1) and combined action of human glioma U251 cells for 24h were 72.43%, 75.56% and 23.45%, respectively. The apoptotic rates of glioma cells were 25.61%, 20.59% and 62.33%, respectively, with or without combination of TRAIL (50ng · ml-1) and CDDP (4μg · ml-1). There was a significant difference between the single application group and the combination group (P <0.01). The results of Western blot showed that the expression of DR5 was significantly increased after the treatment with CDDP, and the expression of DR5 gradually increased with the increase of CDDP dose. Conclusion The combined effect of TRAIL and CDDP can significantly enhance the apoptosis of human U251 glioma cells. The possible mechanism is that CDDP may up-regulate the expression of TRAIL death receptor DR5.