Inhibition of autophagy by 3-MA promotes hypoxia-induced apoptosis in human colorectal cancer cells
Inhibition of autophagy by 3-MA promotes hypoxia-induced apoptosis in human colorectal cancer cells
复制标题
3-MA抑制自噬促进缺氧诱导的人结直肠癌细胞凋亡
DOI：
10.26355/eurrev_201902_16992
复制
发表时间：
2019-02-01
影响因子：
3.3
通讯作者：
Yang, X. -D.
中科院分区：
文献类型：
--
作者：
Dong, Y.;Wu, Y.;Yang, X. -D.
目的：在不同类型的人类癌症中，细胞自噬降低了癌细胞对治疗试剂的敏感性。因此，本研究以人结肠癌HCT116细胞为研究对象，探讨3-甲基腺嘌呤(3-MA，一种自噬抑制剂)对体外培养的人结直肠癌细胞自噬的抑制作用是否能增强缺氧诱导的细胞凋亡。缺氧。或3-MA加低氧。还有自噬。观察HCT116细胞的增殖和凋亡情况。Western印迹分析检测自噬特异性蛋白微管相关蛋白轻链3(Lc3)的表达。用流式细胞仪(JC-1染色检测线粒体膜电位)和膜联蛋白V-碘化丙啶(PI)染色检测细胞凋亡。结果：体外培养的HCT116细胞单独缺氧时自噬增加，细胞凋亡率增加，而3-MA和缺氧联合作用明显抑制缺氧诱导的自噬。结论：自噬可能是低氧处理的结肠癌细胞的一种自我防御机制，其抑制可能是结肠癌辅助化疗的一种有前景的策略。
OBJECTIVE: Cell autophagy reduces the sensitivity of cancer cells to therapeutic reagents in various types of human cancer. Therefore, the aim of our study was to use human colorectal cancer HCT116 cells to explore whether inhibition of autophagy by 3-Methyladenine (3-MA, an autophagy inhibitor) is able to enhance hypoxia-induced apoptosis in vitro.MATERIALS AND METHODS: HCT116 cells were treated with 3-MA. hypoxia. or 3-MA plus hypoxia. and the autophagy. apoptosis and proliferation of the HCT116 cells were investigated. Western blot analysis was used to detect autophagy specificity protein microtubule-associated protein light chain 3 (LC3) expression. Effects on apoptosis were evaluated by using flow cytometry (JC-1 staining to measure mitochondrial membrane potential) and annexin V-propidium iodide (PI) staining.RESULTS: The results showed that the treatment of HCT116 cells in vitro with hypoxia alone increased autophagy as well as apoptosis, whereas combination treatment with 3-MA and hypoxia markedly inhibited hypoxia-induced autophagy. but increased hypoxia-induced cell apoptosis.CONCLUSIONS: Autophagy might play a role as a self-defense mechanism in hypoxia-treated colon cancer cells, and its inhibition could be a promising strategy for the adjuvant chemotherapy of colon cancer.