[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AnnexinVFITC-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48hKYSE-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。
[Purpose ] To investigate the effect of histone deacetylases inhibitor trichostatin A (TSA) and sodium butyrate (NAB) on apoptosis of human KYSE-150 esophageal carcinoma cells. [Methods] IC50 value and eytotoxity of KYSE-150 cells were detected by MTT method. Apoptotic cells and cell cycle distribution were analyzed by FCM using Annexin V FITC-PI and PI staining methods respectively. Expressions of p21 and Bmi-I were evaluated by Western blot method. DNA damage was assayed by γ-H2AX staining.[Resuhs] TSA and NaB significantly inhibited the proliferation of KYSE-150 cells in a dose-dependent manner,with IC50 0.55μmol/L and 5.6mmol/L respectively after 48h. Flow cytometric analysis showed that TSA and NaB induced apoptosis and arrested KYSE-150 cells in the G2/M phase. In addition,p21 protein expression in KYSE-150 cells and DNA damage increased after exposure to TSA and NaB,while Bmi-1 expression was down-regulated. [Conclusion] TSA and NaB could induce apoptosis of human KYSE-150 esophageal carcinoma cells through inhibiting Bmi-1 expression,en- haneement of p21 expression and G2/M arrest,and affect the ability to repair DNA damage.