论文部分内容阅读
[目的]研究组蛋白去乙酰化酶抑制剂曲古菌素A和丁酸钠诱导人食管鳞癌细胞KYSE-150凋亡的作用及机理。[方法]MTT法测IC50值及细胞毒作用,流式细胞仪Annexin V FITC-PI法检测细胞凋亡发生率,PI染色法检测细胞周期变化,蛋白印迹法检测细胞p21和Bmi-1蛋白表达情况,γ-H2AX荧光染色检测细胞DNA损伤。[结果]曲古菌素A和丁酸钠能抑制人食管鳞癌细胞KYSE-150生长且呈浓度依赖关系,作用48h KYSE-150细胞IC50值分别为0.55μmol/L和5.6mmol/L;曲古菌素A和丁酸钠能诱导细胞凋亡,使细胞周期阻滞于G2/M期,同时细胞p21蛋白表达增高,Bmi-1蛋白表达降低,DNA损伤增强。[结论]组蛋白去乙酰化酶抑制剂曲古菌素A和丁酸钠能抑制Bmi-1表达从而激活p21蛋白表达,诱导人食管鳞癌细胞KYSE-150细胞周期阻滞、DNA损伤和凋亡。
[Objective] To investigate the effect and mechanism of histone deacetylase inhibitor trichostatin A and sodium butyrate on apoptosis of human esophageal squamous carcinoma cell line KYSE-150. [Methods] The IC50 value and cytotoxicity were measured by MTT assay. The apoptosis rate was detected by flow cytometry Annexin V FITC-PI and the cell cycle was detected by PI staining. The expression of p21 and Bmi-1 protein was detected by Western blotting The situation, γ-H2AX fluorescence staining of DNA damage. [Result] Trichostatin A and sodium butyrate inhibited the growth of human esophageal squamous cell carcinoma KYSE-150 in a concentration-dependent manner. The IC50 values of KYSE-150 cells were 0.55 μmol / L and 5.6 mmol / L respectively after 48 hours of treatment. Both arginin A and sodium butyrate can induce cell apoptosis, arrest the cell cycle in G2 / M phase, increase p21 protein expression, decrease Bmi-1 protein expression and increase DNA damage. [Conclusion] The histone deacetylase inhibitors, trichostatin A and sodium butyrate, can inhibit the expression of Bmi-1, activate the expression of p21 and induce cell cycle arrest, DNA damage and apoptosis in human esophageal squamous cell carcinoma KYSE-150 cells Death.