低氧微环境下自噬对顺铂处理的肺腺癌A549细胞增殖活性的影响

The effect of autophagy on the proliferation activity of human adenocarcinoma A549 cell treated with DDP in hypoxia condition

目的观察在低氧环境下自噬对人肺腺癌A549细胞化疗敏感性的影响。方法常规低氧培养人肺腺癌A549细胞,分为低氧对照组及低氧+3-甲基腺嘌呤(3-MA)组,分别采用透视电子显微镜观察不同时间点细胞自噬体的变化,Western blot检测自噬标记蛋白微管相关蛋白1轻链3(LC3)蛋白表达,分析LC3II/LC3I比值变化。随后每组再给予顺铂(DDP)处理后,采用MTT比色法检测细胞增殖活性。结果低氧状态下,A549细胞自噬体形成增加,LC3II/LC3I蛋白比值表达升高,加入自噬抑制剂3-MA后,自噬体数量、LCII/LCI比值较单纯低氧组均下降。予顺铂处理后,低氧+3-MA组细胞增殖活性较低氧组下降。结论在低氧环境中,A549细胞保护性自噬增强,抑制自噬可增强A549细胞对顺铂化疗的敏感性。

Objective To observe the effect of autophagy on the chemosensitivity of human adenocarcinoma A549 cell in hypoxia condition.Methods Culture A549 cells in hypoxia condition,then divided them into two groups：hypoxia group and 3-MA plus hypoxia group,electron microscopy was used to detect autophagosme levels at differerent time points,the expression of LC3 was examined by western blot.The proliferation activity of A549 cells after DDP treatment was determined by MTT assay.Results The formation of autophagosome and the ratio of LC3II/LC3I were increased in hypoxia condition,after treatment with autophagy inhibitor 3MA,the quantity of autophagosome and the ration of LC3II/LC3I were decreased.The proliferation activity of 3-MA plus hypoxia group was higher than hypoxia group after treatment with DDP.Conclusion The level of protective autophagy of A549 cell was increased in hypoxia condition,inhibit autophagy can improve the chemosensitivity of A549 cell to DDP.