花旗松素通过PI3K/AKT/mTOR通路对宫颈癌SiHa细胞自噬、凋亡和衰老的影响

Effects of taxifolin on autophagy,apoptosis and senescence of SiHa cells in cervical cancer through PI3K/AKT/mTOR pathway

关键词:  宫颈癌  花旗松素  自噬  凋亡

Key words:  cervical cancer  taxifolin  autophagy  apoptosis

摘要:

[目的] 观察花旗松素（Taxifolin）对宫颈癌SiHa细胞自噬，凋亡和衰老的影响。[方法] 采用0、5、10、20μmol/LTaxifolin处理宫颈癌SiHa细胞。CCK-8法检测细胞细胞增殖倍数；蛋白免疫印迹检测Beclin1、p62、LC3II/LC3I蛋白表达水平；免疫荧光检测LC3+含量；流式细胞仪检测细胞凋亡；流式分选检测线粒体膜电位；蛋白免疫印迹检测Bcl-2、Bax、Caspase-3、cleavedCaspase-3、Caspase-9、cleavedCaspase-9、磷脂酰肌醇-3-激酶（PI3K）、pPI3K、蛋白激酶B（AKT）、p-AKT、哺乳动物雷帕霉素靶蛋白（mTOR）、p-mTOR蛋白表达水平，加入AKT激活剂SC79进行验证。[结果] 与Taxifolin0μmol/L组相比较，Taxifolin10、20μmol/L组细胞增殖倍数显著降低（P<0.05），p62蛋白水平显著降低（P<0.05），Beclin1、LC3II/LC3I蛋白水平显著升高（P<0.05），LC3+含量显著升高（P<0.05），凋亡率显著升高（P<0.05），线粒体膜电位显著升高（P<0.05），Bcl-2/Bax比值显著降低（P<0.05），cleavedCaspase-3/Caspase-3、cleavedCaspase-9/Caspase-9比值升高（P<0.05），p-PI3K/PI3K、p-AKT/AKT、p-mTOR/mTOR比值显著降低（P<0.05）。加入PI3K/AKT信号通路激活剂SC79可逆转花旗松素对SiHa细胞凋亡、自噬及PI3K/AKT信号通路相关蛋白表达的影响。[结论] Taxifolin通过抑制PI3K/AKT/mTOR通路的活化诱导宫颈癌SiHa细胞自噬，凋亡和衰老。

Abstract:

[Objective] To observe the effects of taxifolin on autophagy,apoptosis and senescence of SiHa cells in cervical cancer. [Methods] SiHa cells of cervical cancer were treated with 0,5,10 and 20 μmol/L Taxifolin. Cell proliferation multiple was measured by CCK-8 method. Beclin1,p62 and LC3II/LC3I were detected by western blot. LC3 + was detected by immunofluorescence assay. Flow cytometry was used to detect apoptosis. Mitochondrial membrane potential was detected by flow sorting. Protein expression levels of Bcl-2,Bax,Caspase-3,cleaved Caspase-3,Caspase-9,cleaved Caspase-9,PI3K,p-PI3K,AKT,p-AKT,mTOR and p-mTOR were detected by protein western blot,AKT activator SC79 was added for verification. [Results] Compared with taxifolin 0 μmol/L group,10,20 μmol/L taxifolin group cell proliferation ratio decreased significantly (P<0.05),p62 protein levels decreased significantly (P<0.05),Beclin1,LC3II/LC3I protein were significantly increased (P<0.05),LC3+ content increased significantly(P<0.05),the apoptosis rate increased significantly(P<0.05),significantly increased mitochondrial membrane potential (P<0.05),the Bcl-2/Bax ratio decreased significantly (P<0.05),The ratio of cleaved Caspase-3/Caspase-3 and cleaved Caspase-9/Caspase-9 was increased (P<0.05),and the ratio of p-PI3K/PI3K,p-AKT/AKT, and p-mTOR/mTOR were significantly decreased(P<0.05). The addition of SC79,a PI3K/Akt signaling pathway activator, could reverse the effects of taxifolin on apoptosis,autophagy and PI3K/AKT signaling pathway related protein expression in SiHa cells. [Conclusion] Taxifolin induces autophagy,apoptosis and senescence of SiHa cells in cervical cancer by inhibiting the activation of PI3K/AKT/mTOR pathway.