摘要
背景与目的:使用分子靶向药物治疗胶质瘤是神经肿瘤领域的研究热点。本文探讨组蛋白去乙酰化酶抑制剂MS-275对脑胶质瘤细胞株U251细胞增殖和迁移的影响及其机制。方法:以不同药物浓度梯度与U251细胞分别共培养24、48和72h后,应用CCK-8法检测肿瘤细胞增殖,Transwell法检测药物作用前后U251细胞迁移能力的变化,Annexin V-FITC/PI法经流式检测细胞凋亡率,实时定量PCR检测IκB-α的RNA含量,免疫印迹检测IκB-α蛋白、磷酸化IκB-α蛋白和聚ADP核糖聚合酶(PARP)表达。结果:MS-275能显著抑制U251细胞的增殖。MS-275药物浓度40nmol/mL,与U251共培养72h,细胞增殖抑制率为(79.0±1.7)%;经药物作用72h后,U251细胞凋亡率为(29.000±2.306)%;实时定量PCR检测IκB-α的RNA含量随药物作用时间的延长而降低,免疫印迹检测提示IκB-α蛋白表达水平降低,IκB-α蛋白磷酸化被抑制,PARP被剪切。结论:MS-275对胶质瘤细胞的增殖和迁移抑制作用具有浓度和时间依赖性,其机制是通过抑制IκB-α蛋白磷酸化诱导凋亡实现的。
BACKGROUND OBJECTIVE： Employment of targeting therapy in the treatment of glioma is a hot topic in neuro-oncology.In this study,we explored effect and mechanism of MS-275 on the proliferation and migration of brain glioma cells.METHODS： U251 cells were respectively cultured for 24,48 and 72h,with different concentrations of MS-275.CCK-8 assay was used to observe the proliferation of U251 cells after MS-275 treatment.The migration of U251 cells treated with or without MS-25 was measured with Transwell assay.Apoptosis induced by MS-275 in U251 was detected with flow cytometry using annexin V-FITC/PI.RNA level of IκB-α was examined with real-time PCR.The expression of total and phosphorylated protein of IκB-αand PARP were detected with Western blotting.RESULTS： MS-275 significantly inhibited the proliferation of U251.After the treatment of MS-275 at the concentration of 40 nmol/ml for 72 h,the proliferation inhibition rate of U251 was（79.0±1.7）%.MS-275 induced apoptosis in U251.Apoptosis rate was（29.000±2.306） % in U251 after 72h treatment of MS-275.RNA level of IκB-αin U251 reduced after MS-275 treatment.The expression of IκB-αprotein was downregulated and phosphorylated IκB-α decreased greatly,with the cleavage of PARP detected by Western blot,after MS-275 treatment.CONCLUSON： MS-275 supressess proliferation and migration,and induces apoptosis in U251 cells,of U251 cellsthrough the inhibition of the phosphorylation of IκB-αprotein.
出处
《中国神经肿瘤杂志》
2011年第2期82-87,共6页
Chinese Journal of Neuro-Oncology
