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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 & AIM: The use of molecular targeted drugs in the treatment of glioma is a hot topic in the field of neuro-tumor. This article discusses the histone deacetylase inhibitor MS-275 on glioma cell line U251 cells proliferation and migration and its mechanism. Methods: The proliferation of U251 cells was detected by CCK-8 assay after 24, 48 and 72 hours, respectively. The migration ability of U251 cells was detected by Transwell assay. The Annexin V-FITC / PI assay The apoptosis rate of IκB-α was detected by flow cytometry and the content of IκB-α was detected by real-time quantitative PCR. The expression of IκB-α protein, phosphorylated IκB-α protein and poly ADP ribose polymerase (PARP) were detected by Western blotting. Results: MS-275 significantly inhibited the proliferation of U251 cells. MS-275 drug concentration of 40nmol / mL, co-cultured with U251 72h, cell proliferation inhibition rate was (79.0 ± 1.7)%; 72h after treatment, U251 cells apoptosis rate was (29.000 ± 2.306)%; The RNA content of IκB-α decreased with the prolongation of the drug duration. Immunoblotting indicated that the expression of IκB-α was decreased, phosphorylation of IκB-α was inhibited and PARP was cleaved. CONCLUSION: The inhibitory effect of MS-275 on the proliferation and migration of glioma cells is concentration-dependent and time-dependent, and its mechanism is through the inhibition of IκB-α protein phosphorylation induced apoptosis.