抑制自噬对X射线诱导结肠癌HCT116细胞增殖与凋亡的影响

Effect of Autophagy Inhibition on X-ray Induced Proliferation and Apoptosis of Colon Cancer HCT116 Cells

目的探讨自噬抑制剂3-甲基腺嘌呤(3-MA)与X射线联合应用对人结肠癌细胞HCT116增殖和凋亡的影响。方法体外培养HCT116细胞,分为对照组、X射线组(6Gy)、3-MA组(5mmol·L-1)以及X射线联合3-MA作用组。CCK-8检测细胞相对存活率,流式细胞术检测细胞凋亡率,Western blot检测自噬标志物p62和LC3 II/LC3 I、凋亡家族Bax和Bcl-2的蛋白表达。结果X射线照射HCT116细胞后,细胞增殖活性下降,凋亡率升高(P<0.01),并且可诱导细胞产生自噬。3-MA和X射线联合作用较单独X射线照射细胞的增殖活性降低,细胞自噬水平下降,凋亡率升高(P<0.01)。结论X射线在诱导HCT116细胞凋亡的同时可诱导自噬,通过3-MA抑制自噬可增强HCT116细胞对X射线作用的敏感性。

Objective To investigate the effect of autophagy inhibitor 3-methyladenine(3-MA)combined with X-ray on proliferation and apoptosis of human colon cancer cell line HCT116.Methods HCT116 cells were cultured in vitro and divided into control group,X-ray group(6Gy),3-MA group(5mmol·L-1)and X-ray combined with 3-MA treatment group.The relative survival rate of cells was detected by CCK-8,the apoptosis rate was detected by flow cytometry,and the expression of autophagy markers p62 and LC3 II/LC3 I,apoptosis family Bax and Bcl-2 were detected by Western blot.Results After the treatment of X-ray,the cell proliferation activity decreased(P<0.01),the apoptosis rate increased(P<0.01),and autophagy could be induced.The combined activity of 3-MA and X-ray was significantly lower than that of X-ray irradiated cells(P<0.01),the level of autophagy decreased,and the apoptosis rate increased significantly(P<0.01).Conclusion X-ray can induce autophagy while inducing apoptosis of HCT116 cells,and autophagy inhibition by 3-MA can enhance the sensitivity of HCT116 cells to X-rays.