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目的组蛋白的异常修饰(去乙酰化)与胶质瘤的发生和发展关系密切,本研究的目的是探讨组蛋白去乙酰化酶抑制剂曲古霉素A(trichomycin A,TSA)对胶质瘤细胞系凋亡的调节作用。方法将浓度分别为0.2μmol/L、0.4μmol/L和0.8μmol/L的TSA加入胶质瘤细胞系U251细胞中,应用Westernblot法检测U251细胞系经不同浓度的TSA处理后细胞内胱氨酸蛋白酶-3(caspase-3),乙酰化组蛋白H3和H4的表达,应用流式细胞术(Annexin V-FITC染色)检测不同药物浓度处理组中U251细胞的凋亡率。结果 TSA处理后,U251细胞内乙酰化组蛋白H3、H4及caspase-3的蛋白水平明显上调(P<0.01),具有明显的剂量依赖性。流式细胞术结果显示U251细胞TSA处理后细胞的凋亡率具有剂量依赖性显著上调。结论 TSA剂量依赖性上调胶质瘤细胞系U251中caspase-3表达,促进细胞凋亡。
The purpose of this study is to investigate the effect of histone deacetylase inhibitor trichomycin A (TSA) on the glioma development and progression. Regulatory effect of tumor cell apoptosis. Methods TSA of 0.2μmol / L, 0.4μmol / L and 0.8μmol / L were added to glioma cell line U251. Western blot was used to detect the cystine content of U251 cells treated with different concentrations of TSA The expressions of caspase-3, acetylated histone H3 and H4 were detected by flow cytometry (FCM). The apoptosis rate of U251 cells in different drug concentration groups was detected by flow cytometry (Annexin V-FITC staining). Results After TSA treatment, the protein levels of acetylated histone H3, H4 and caspase-3 in U251 cells were significantly up-regulated (P <0.01) in a dose-dependent manner. Flow cytometry results showed that apoptosis rate of U251 cells after TSA treatment significantly increased in a dose-dependent manner. Conclusion TSA dose-dependently upregulates caspase-3 expression in glioma cell line U251 and promotes apoptosis.