抑制自噬增强盐酸阿霉素诱导的人结直肠癌细胞凋亡

Inhibition of autophagy enhances apoptosis induced by doxorubicin hydrochloride in human colon cancer cells

目的 探讨盐酸阿霉素(DOX)抑制结直肠癌HT29、 HCT15细胞增殖的可能机制。方法 HT29、 HCT15细胞采用(0、 0.08、 0.16、 0.32、 0.64、 1.28)μmol/L DOX处理24、 48、 72 h,CCK-8法检测细胞增殖活性确定DOX最佳处理浓度和处理时间。将HT29、 HCT15细胞分别设为对照组(只加DMSO处理)和(0.16、 0.32、 0.64、 1.28)μmol/LDOX处理组,Western blot法检测抑制自噬对凋亡的影响时添加3-甲基腺嘌呤(3-MA)组、 3-MA联合DOX组。集落形成实验检测结直肠癌细胞的集落形成能力,Western blot法检测细胞B细胞淋巴瘤因子2(Bcl2)、 Bcl2相关X蛋白(BAX)、 beclin 1、微管相关蛋白1轻链3(LC3)的蛋白表达。结果 DOX抑制结直肠癌细胞的增殖和集落形成,促进细胞发生凋亡,呈浓度依赖性;DOX促进细胞发生自噬,beclin 1和LC3Ⅱ表达增加,呈浓度依赖性;抑制自噬后可增加DOX对结直肠癌细胞的促凋亡作用。结论 DOX抑制结直肠癌细胞增殖并促进其凋亡且抑制自噬有促凋亡作用。

Objective To investigate the possible mechanism of doxorubicin hydrochloride(DOX) inhibiting the proliferation of HT29 and HCT15 colon cancer cells. Methods The gradient concentrations of(0, 0.08, 0.16, 0.32, 0.64, 1.28) μmol/L DOX were used to treat HT29 and HCT15 cells for 24, 48 and 72 hours, and the cell proliferation activity was detected by CCK-8 assay to determine the optimal DOX concentration and treatment time. According to different treatments, HT29 and HCT15 cells were divided into 2 groups: control group(only DMSO treatment) and(0.16, 0.32, 0.64, 1.28) μmol/L DOX group. Western blot was used to detect the effect of inhibiting autophagy on apoptosis, with 3-methyladenine(3-MA) group and 3-MA combined with DOX group supplemented. The colony formation of colon cancer cells was detected by colony formation assay, and the expression of cell B-cell lymphoma 2(Bcl2), Bcl2-associated X protein(BAX), beclin 1, and LC3 protein were detected by Western blot. Results DOX inhibited the proliferation and colony formation of colon cancer cells, and promoted cell apoptosis in a concentration-dependent manner;DOX promoted autophagy in cells, and the expression of beclin 1 and LC3Ⅱ increased in a concentration-dependent manner;DOX promoted apoptosis of colon cancer cells, which was improved by inhibiting autophagy. Conclusion DOX inhibits the proliferation of colon cancer cells and promotes their apoptosis, and inhibition of autophagy in colon cancer cells can increase the sensitivity of apoptosis induced by DOX.