摘要
目的探讨Survivin对体外培养的人脑胶质瘤细胞(U251)替莫唑胺化疗敏感性的影响。方法将人脑胶质瘤细胞U251中Survivin的表达上调或者抑制后,经替莫唑胺处理。采用四甲基偶氮唑蓝(MTT)比色法,平板克隆形成试验评价各组细胞增殖能力,流式细胞仪检测细胞凋亡,蛋白免疫印迹法检测各组细胞激活caspase-3的表达情况。结果替莫唑胺处理后,与亲代细胞相比,下调Survivin的表达,细胞的生长抑制效果增强,凋亡率增加,激活的caspase-3蛋白表达增多;上调Survivin的表达,细胞生长抑制作用减弱,凋亡率减少,激活的caspase-3蛋白表达下降。结论 Survivin可通过抑制凋亡,影响胶质瘤细胞对替莫唑胺的敏感性。
Objective The effect of Survivin on teroozolomide (TMZ) resistance of U251 glioma cells is discussed. Methods The Survivin RNA plasmids or the specific shRNA vector targeting Sundvin was transfected into U251 cells, respectively. The cellular growth activity was assayed by methyl thiazolyltetrazolium (MTT). Cell surviving fraction was calculated by colony forming assay. The apoptosis was assayed by the Annexin V assay by flow cytometer (FCM). The expression of activated caspase-3 was detected by western blot. Results After treated with TMZ, compared with parent cells, cell proliferation was inhibited significantly, apoptosis rate was increased, and the expression of activated caspase-3 protein was promoted in down-regulation Survivin group; while in up-regulation Survivin group, cell proliferation was promoted, apoptosis rate was decreased, and the expression of activated caspase-3 protein was declined. Conclusion Survivin can suppress TMZ sensitivity of glioma cells by inhibiting the apoptosis in vitro.
出处
《中华神经外科疾病研究杂志》
CAS
2016年第4期317-320,共4页
Chinese Journal of Neurosurgical Disease Research