白藜芦醇经PI3K/Akt/mTOR信号通路诱导人肾癌786-O细胞自噬

白藜芦醇(resveratrol)可抑制人肾癌786-O细胞增殖,并诱导其凋亡,但是白藜芦醇对786-O细胞自噬(autophagy)的影响及机制尚不清楚。为探究其机制,体外培养786-O细胞,采用CCK-8检测786-O细胞活力;TUNEL染色检测786-O细胞凋亡;透射电子显微镜观察786-O细胞自噬体;吖啶橙染色观察786-O细胞自噬小泡;GFP-LC3质粒转染分析观察786-O细胞自噬体;Western印迹检测LC3、beclin-1、PI3K、p-PI3K、Akt、p-Akt、mTOR和p-mTOR的表达。结果显示,白藜芦醇以浓度和时间依赖性的方式抑制786-O细胞活力,并诱导细胞凋亡;与对照组相比,白藜芦醇使786-O细胞自噬增强;Western印迹结果显示,与对照组相比,白藜芦醇组LC3-II/LC3-I和Beclin-1显著增高(P<0.01),表明白藜芦醇导致786-O细胞自噬体积累。与对照组相比,白藜芦醇使786-O细胞的p-PI3K/PI3K,p-Akt/Akt和p-mTOR/mTOR显著降低(P<0.01),表明白藜芦醇可通过PI3K/Akt/mTOR信号通路增强自噬。综上所述,白藜芦醇通过抑制PI3K/Akt/mTOR信号通路从而诱导786-O细胞自噬。

Resveratrol Induces Autophagy in 786-O Cells of Human Renal Carcinoma via the PI3K/Akt/mTOR Signaling Pathway

Resveratrol inhibits the proliferation and induces apoptosis of human renal carcinoma 786-O cells.However,the effect and mechanism of resveratrol on autophagy in 786-O cells remain unclear.To explore its mechanism,786-O cells were cultured in vitro.First,CCK-8 assay results showed that resveratrol inhibited cell viability in a dose-and time-dependent manner.Secondly,TUNEL staining revealed that resveratrol treatment induces apoptosis.Thirdly,we measured autophagy levels using a combination of transmission electron microscopy,acridine orange staining as well as GFP-LC3 plasmid transfection analysis.And the autophagy levels in the resveratrol group increased significantly compared with the control group.Lastly,we directly measured the expressions of LC3,Beclin1,PI3 K,p-PI3 K,Akt,p-Akt,mTOR and p-mTOR by Western blotting,and found that LC3-II/LC3-I and Beclin-1 in the resveratrol group were significantly higher than those in the control group(P<0.01),while p-PI3 K/PI3 K,p-Akt/Akt and p-mTOR/mTOR were reduced(P<0.01).indicating that resveratrol caused accumulation of autophagosomes in 786-O cells.In conclusion,we propose that resveratrol induces autophagy in 786-O cells by inactivating the PI3 K/Akt/mTOR signaling pathway.