PPVI通过调节PI3K/AKT/mTOR通路对食管癌细胞ECA-109凋亡和自噬的影响

Effects of PPVI on Apoptosis and Autophagy of Esophageal Cancer Cells ECA-109 by Regulating PI3K/AKT/mTOR Pathway

目的:探讨中药提取物重楼皂苷VI(PPVI)通过调节PI3K/AKT/mTOR通路对食管癌细胞凋亡和自噬的影响。方法:将对数生长期ECA-109细胞分为对照组(不进行处理)、PPVI低剂量组(添加2.5mg/L的PPVI处理)、PPVI中剂量组(添加5.0mg/L的PPVI处理)、PPVI高剂量组(添加7.5mg/L的PPVI处理)、IGF-1组(添加10nmol/mL的IGF-1处理)、IGF-1+PPVI组(添加10nmol/mL的IGF-1和7.5mg/L的PPVI处理),CCK-8检测ECA-109细胞活力,Transwell检测ECA-109细胞迁移,流式细胞术检测ECA-109细胞凋亡,Western blot检测PI3K/AKT/mTOR信号通路和自噬相关蛋白。结果:与对照组相比,PPVI低剂量组、PPVI中剂量组、PPVI高剂量组中ECA-109细胞活力、迁移能力明显降低,凋亡能力明显升高(P<0.001),而IGF-1组ECA-109细胞活力、迁移能力显著升高,凋亡能力明显降低(P<0.001);与PPVI高剂量组相比,IGF-1+PPVI组的ECA-109细胞活力、迁移能力明显升高,凋亡能力明显降低(P<0.001);与对照组相比,PPVI低剂量组、PPVI中剂量组、PPVI高剂量组ECA-109细胞中p-PI3K、p-Akt、p-mTOR、LC3BⅡ/Ⅰ蛋白水平明显降低,p62蛋白水平明显升高(P<0.001),而IGF-1组ECA-109细胞中p-PI3K、p-Akt、p-mTOR、LC3BⅡ/Ⅰ蛋白水平显著升高,p62蛋白水平明显降低(P<0.001);与PPVI高剂量组相比,IGF-1+PPVI组的ECA-109细胞中p-PI3K、p-Akt、p-mTOR、LC3BⅡ/Ⅰ蛋白水平明显升高,p62蛋白水平明显降低(P<0.001)。结论:PPVI可能通过抑制PI3K/AKT/mTOR通路诱导ECA-109细胞凋亡和自噬,抑制ECA-109细胞活力和迁移。

Objective:To investigate the effects of saponin VI(PPVI)on apoptosis and autophagy of esophageal cancer cells by regulating PI3K/AKT/mTOR pathway.Methods:ECA-109 cells were divided into control(no treatment),PPVI low-dose group(PPVI treatment added 2.5mg/L),PPVI medium-dose group(PPVI treatment added 5.0mg/L),PPVI high-dose group(PPVI treatment added 7.5mg/L),IGF-1 group(IGF-1 added 10 mg/L)nmol/mL (IGF-1 treatment),IGF-1+PPVI group(10 nmol/mL IGF-1 and 7.5mg/L PPVI treatment were added),CCK-8 was used to detect ECA-109 cell viability,Transwell was used to detect ECA-109 cell migration.Apoptosis of ECA-109 cells was detected by flow cytometry,and PI3K/AKT/mTOR signaling pathway.Results:The proliferation activity of ECA-109 cells in low-dose PPVI,medium-dose PPVI and high-dose,and the apoptosis increased(P<0.001),while the proliferation activity and migration ECA-109 cells in IGF-1 increased.The apoptosis capacity was significantly decreased(P<0.001).Compared with PPVI high-dose,proliferation activity and migration ability of ECA-109 cells in IGF-1+PPVI increased,and apoptosis ability was significantly decreased(P<0.001).The protein levels of P-PI3K,P-Akt,P-mTOR and LC3BⅡ/Ⅰin ECA-109 cells of PPVI low,PPVI medium and PPVI high decreased,while the protein p62 were significantly increased(P<0.001).The protein levels of P-PI3K,P-Akt,P-mTOR and LC3BⅡ/Ⅰin ECA-109 cells of IGF-1 were significantly increased,while p62 decreased(P<0.001).PPVI high-dose,the protein levels of P-PI3K,P-Akt,P-MTOR and LC3BⅡ/Ⅰin ECA-109 cells of IGF-1+PPVI increased,while the protein p62 were significantly decreased(P<0.001).Conclusions:PPVI may induce apoptosis and autophagy of ECA-109 cells by inhibiting PI3K/AKT/mTOR pathway,and inhibit proliferation and migration of ECA-109 cells.