芹菜素通过AMPK/mTOR通路调控肺癌A549细胞自噬

目的 探讨芹菜素（APG）对肺癌A549细胞增殖及自噬的影响及其与AMPK/mTOR通路的关系。方法 体外培养肺癌A549细胞，采用CCK-8法检测不同浓度芹菜素处理A549细胞不同时间后对细胞增殖的影响。后续实验分为空白对照组(0 μmol/L APG)，低、中、高浓度实验组和抑制剂组5组，采用MDC染色法检测各组对细胞自噬的影响，免疫印迹法检测自噬相关蛋白以及AMPK/mTOR通路相关蛋白表达情况。结果 芹菜素可显著抑制A549细胞增殖，且呈浓度依赖性（P<0.05）；20、40、80 μmol/L芹菜素组随作用时间延长，对A549细胞的增殖抑制率逐渐增加，呈时间依赖关系（P<0.05）。与空白对照组比较，实验组A549细胞中含高亮点状荧光的自噬小泡数量及IOD值逐渐增加（P<0.05），芹菜素可浓度依赖性上调LC3Ⅰ、LC3Ⅱ、p-AMPKα蛋白表达水平及p-AMPKα/AMPKα比值（P<0.05），下调p-mTOR、p62蛋白表达水平及p-mTOR/mTOR比值（P<0.05）。与高浓度实验组比较，抑制剂组细胞中含高亮点状荧光的自噬小泡减少，IOD值下降，LC3Ⅰ、LC3Ⅱ、p-AMPKα蛋白表达水平及p-AMPKα/AMPKα比值降低，而P62、p-mTOR蛋白表达水平及p-mTOR/mTOR比值升高（P<0.05）。结论 芹菜素能抑制肺癌A549细胞增殖，诱导其自噬，作用机制可能与AMPK/mTOR通路有关

Apigenin regulates autophagy of lung cancer A549 cells via AMPK/mTOR pathway

Objective To study the effect of apigenin on the proliferation and autophagy of lung cancer A549 cells and its relationship with the AMPK/mTOR pathway. Methods Lung cancer A549 cells were cultured in vitro. The effect of different concentrations of apigenin on cell proliferation after treatment of A549 cells for different times was detected by CCK-8 method. The follow-up experiment was divided into five groups: blank control group, low, medium and high concentration experimental groups and inhibitor group. MDC staining was used to detect the effect of each group on cellular autophagy. The expression of autophagy-related proteins and AMPK/mTOR pathway-related proteins were detected by Western blotting. Results Apigenin significantly inhibited the proliferation of A549 cells in a concentration-dependent manner (P<0.05). The proliferation inhibition rate of A549 cells in the 20, 40 and 80 μmol/L apigenin groups gradually increased with increasing duration of action in a time-dependent manner(P<0.05). The number of autophagic vesicles containing high bright spot fluorescence and IOD values in A549 cells gradually increased in the experimental groups compared with the blank control group(P<0.05). Apigenin could up-regulate LC3Ⅰ, LC3Ⅱ, p-AMPKα protein expression levels and p-AMPKα/AMPKα ratio in a concentration-dependent manner (P<0.05), and down-regulate p-mTOR, p62 protein expression levels and p-mTOR/mTOR ratio (P<0.05). Compared with the high concentration experimental group, the autophagic vesicles containing high bright fluorescence, the IOD values, the LC3Ⅰprotein expression level, the LC3Ⅱ level, the p-AMPKα level, and the p-AMPKα/AMPKα ratio were significantly decreased in the inhibitor group(P<0.05). The P62 protein expression level, p-mTOR level, and p-mTOR/mTOR ratio were significantly increased(P<0.05). Conclusion Apigenin inhibits the proliferation and induces autophagy in A549 cells, and the mechanism of action may be related to the AMPK/mTOR pathway