茶黄素调节PI3K/Akt/mTOR信号通路对A549细胞增殖凋亡及自噬的影响  

Effects of Theaflavin on Proliferation Apoptosis and Autophagy of A549 Cell by Regulating the PI3K/Akt/mTOR Signaling Pathway

作　　者：段兴隆 张华 DUAN Xinglong;ZHANG Hua(Hanzhong People's Hospital,Shaanxi Hanzhong 723000,China)

机构地区：[1]陕西省汉中市人民医院呼吸内科,陕西汉中723000

出　　处：《河北医学》2024年第9期1473-1478,共6页Hebei Medicine

基　　金：陕西省汉中市人民医院项目计划,(编号:sy-2023-02)。

摘　　要：目的:探讨茶黄素调节磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(AKT)/哺乳动物雷帕霉素靶蛋白(mTOR)信号通路对A549细胞增殖、凋亡及自噬的影响。方法:测定0、2.5、5、10、20、40、80、160μmoL/L茶黄素干预A549细胞后增殖率,筛选出合适的作用浓度。将A549细胞分为control组、低-茶黄素组(20μmoL/L茶黄素)、中-茶黄素组(40μmoL/L茶黄素)、高-茶黄素组(80μmoL/L茶黄素)、高-茶黄素+740Y-P组(80μmoL/L茶黄素+30μmoL/L的PI3K/Akt/mTOR信号通路激活剂740Y-P)。CCK-8法、平板集落形成实验测定A549细胞增殖;流式细胞术、MDC法分别测定A549细胞凋亡及自噬空泡生成;Western blot测定A549细胞PI3K/Akt/mTOR信号通路蛋白及自噬蛋白表达。结果:与0μmoL/L茶黄素对比,2.5、5、10、20、40、80、160μmoL/L的茶黄素均可抑制A549细胞增殖,经计算茶黄素对A549细胞的IC50值为36.72μmoL/L。与control组比较,低-茶黄素组、中-茶黄素组、高-茶黄素组A549细胞增殖率、集落形成率、p-PI3K/PI3K、p-Akt/Akt、p-mTOR/mTOR均降低,凋亡率、自噬空泡相对含量、LC3II/LC3I、Beclin-1蛋白表达均升高,且均呈茶黄素剂量依赖性变化(P<0.05)。与高-茶黄素组对比,高-茶黄素+740Y-P组A549细胞增殖率、集落形成率、p-PI3K/PI3K、p-Akt/Akt、p-mTOR/mTOR均升高,凋亡率、自噬空泡相对含量、LC3II/LC3I、Beclin-1蛋白表达均降低(P<0.05)。结论:茶黄素可通过抑制PI3K/Akt/mTOR信号通路激活,诱导保护性自噬,进而促进肺癌细胞凋亡,抑制其增殖。Objective:To investigate the effects of theaflavin on A549 cell proliferation,apoptosis,and autophagy by regulating the phosphatidylinositol 3-kinase(PI3K)/protein kinase B(AKT)/mammalian target of rapamycin(mTOR)signaling pathway.Methods:The proliferation rate of A549 cells was measured after intervention with 0,2.5,5,10,20,40,80,and 160μmol/L theaflavin,and the appropriate concentration of action was screened.A549 cells were separated into control group,low theaflavin group(20μmol/L theaflavin),medium theaflavin group(40μmol/L theaflavin),high theaflavin group(80μmol/L theaflavin),and high theaflavin+740Y-P group(80μmol/L theaflavin+30μmol/L PI3K/Akt/mTOR signaling pathway activator 740Y-P).CCK-8 method and plate colony formation experiment were applied to determine the proliferation of A549 cells.Flow cytometry and MDC methods were applied to determine apoptosis and autophagic vacuole formation in A549 cells,respectively.Western blot was applied to determine the expression of PI3K/Akt/mTOR signaling pathway proteins and autophagy proteins in A549 cells.Results:Compared with 0μmol/L of theaflavin,2.5,5,10,20,40,80,and 160μmol/L of theaflavin can inhibit the proliferation of A549 cells.The IC50 value of theaflavin on A549 cells was calculated to be 36.72μmol/L.Compared with the control group,the proliferation rate,colony formation rate,p-PI3K/PI3K,p-Akt/Akt,and p-mTOR/mTOR of A549 cells in the low theaflavin group,medium theaflavin group,and high theaflavin group all reduced,the apoptosis rate,relative content of autophagic vacuoles,LC3II/LC3I,and Beclin-1 protein expression all increased,and all showed dose-dependent changes in theaflavin(P<0.05).Compared with the high theaflavin group,the proliferation rate,colony formation rate,p-PI3K/PI3K,p-Akt/Akt,and p-mTOR/mTOR of A549 cells in the high theaflavin+740Y-P group all increased,the apoptosis rate,relative content of autophagic vacuoles,LC3II/LC3I,and Beclin-1 protein expression all reduced(P<0.05).Conclusion:Theaflavin can induce protective autophagy

关 键 词：茶黄素 磷脂酰肌醇3-激酶/蛋白激酶B/哺乳动物雷帕霉素靶蛋白信号通路 自噬 肺癌 

分 类 号：R734.2[医药卫生—肿瘤]