槲皮素调控AMPK/mTOR通路对肺癌A549细胞自噬的影响  

Effect of quercetin on autophagy of lung cancer A549 cells by regulating AMPK/mTOR pathway

作　　者：文兰香 覃世运[1] 陈丽君[1] WEN Lan-Xiang;QIN Shi-Yun;CHEN Li-Jun(Cancer Center of Hainan Third People′s Hospital,Sanya 572000,China)

机构地区：[1]海南省第三人民医院肿瘤中心,三亚572000

出　　处：《中国免疫学杂志》2020年第19期2375-2379,共5页Chinese Journal of Immunology

基　　金：海南省卫生计生行业科研项目(No.17A200034)。

摘　　要：目的:探讨槲皮素对肺癌细胞自噬的影响以及其租用分子机制。方法:体外培养肺癌A549细胞,分为空白对照组、槲皮素低、中、高各剂量组,通过CCK-8法检测细胞增殖能力,MDC染色法检测细胞自噬形态,Western blot检测自噬相关蛋白LC3Ⅰ、LC3Ⅱ、beclin-1以及AMPK/mTOR通路相关蛋白表达情况。结果:与空白对照组比较,槲皮素低、中、高各剂量组对A549细胞增殖抑制率依次升高,呈浓度依赖关系(P<0.05);MDC染色显示,空白对照组中仅有少量细胞含有高亮点状荧光,作用48 h后槲皮素低、中、高各剂量组A549细胞中含高亮点状荧光的自噬泡依次增多;作用48 h后,与空白对照组相比,槲皮素低、中、高各剂量组A549细胞中LC3Ⅱ/LC3Ⅰ、beclin-1蛋白、p-AMPK/AMPK蛋白水平依次升高(P<0.05),p-mTOR/mTOR、p-S6K/S6K蛋白水平依次降低(P<0.05);与槲皮素组相比,槲皮素+AMPK抑制剂组A549细胞中高亮点状荧光的自噬泡减少,p-AMPK/AMPK、LC3Ⅱ/LC3Ⅰ、beclin-1蛋白表达降低(P<0.05),p-mTOR/mTOR、p-S6K/S6K表达升高(P<0.05)。结论:槲皮素可能通过AMPK/mTOR通路,诱导肺癌A549细胞自噬。Objective:To investigate the effect of quercetin on autophagy of lung cancer A549 cells and explore the possible molecular mechanism.Methods:Lung cancer A549 cells were cultured in vitro and divided into blank control group,quercetin low,medium,high dose group cell proliferation was detected by CCK-8 assay,the morphology of autophagy was detected by MDC staining,and the expressions of autophagy-related proteins LC3Ⅰ,LC3Ⅱ,beclin-1 and AMPK/mTOR pathway-related proteins were detected by Western blot.Results:Compared with the blank control group,the inhibition rates of quercetin low,medium,high dose group on A549 cells increased and in a concentration-dependent manner(P<0.05).MDC staining showed that only a few cells in the blank control group contained high bright spot fluorescence.After 48 hours of treatment,the number of autophagic vesicles containing high bright spot fluorescence in A549 cells of quercetin low,medium,high dose groups increased in turn.After 48 hours,compared with the blank control group,the levels of LC3Ⅱ/LC3Ⅰ,beclin-1 protein,p-AMPK/AMPK protein in A549 cells of quercetin low,medium,high dose group increased in turn(P<0.05),while the levels of p-mTOR/mTOR,p-S6 K/S6 K proteins decreased in turn(P<0.05).Compared with quercetin group,the number of autophagic vesicles containing high bright spot fluorescence in A549 cells in quercetin +AMPK inhibitor group decreased,the expressions of p-AMPK/AMPK,LC3Ⅱ/LC3Ⅰ,beclin-1 proteins decreased(P<0.05),and the expressions of p-mTOR/mTOR,p-S6 K/S6 K increased(P<0.05).Conclusion:Quercetin may induce autophagy of lung cancer A549 cells through AMPK/mTOR pathway.

关 键 词：槲皮素 肺癌 自噬 AMPK/mTOR通路 

分 类 号：R734.2[医药卫生—肿瘤]