鱼藤素通过AKT/FoxO1信号通路诱导胃癌细胞凋亡的作用机制

Mechanism of Deguelin in Inducing Gastric Cancer Cell Apoptosis by AKT/FoxO1 Signaling Pathway

目的:研究鱼藤素(De)诱导SGC-7901胃癌细胞凋亡的其作用机制。方法:运用CCK-8细胞活力检测法考察不同浓度(10、20、40、60、80、100 mol/L)鱼藤素作用24、48 h对SGC-7901胃癌细胞的细胞毒性;将SGC-7901胃癌细胞分为对照组及20、40 mol/L鱼藤素药物组,给药作用24 h后,蛋白质印迹法检测p-AKT^(Thr308)、叉头框蛋白O1(FoxO1)、B淋巴细胞瘤-2基因(Bcl-2)等蛋白的表达水平;运用蛋白激酶B(AKT)基因转染使SGC-7901胃癌细胞中AKT过表达,然后给予20、40 mol/L鱼藤素给药作用24 h,以未用AKT基因转染的SGC-7901胃癌细胞作为对照组蛋白质印迹法检测p-AKT^(Thr308)、FoxO1、Bcl-2等蛋白的表达水平,实时荧光定量PCR(RT-qPCR)检测FoxO1、Bcl-2、Bax mRNA的表达水平。结果:不同浓度的鱼藤素对SGC-7901胃癌细胞均具有一定的细胞毒性;20、40 mol/L鱼藤素给药作用24 h能够显著降低SGC-7901胃癌细胞中p-AKT^(Thr308)、Bcl-2等蛋白的表达水平,升高FoxO1的表达水平;与对照组比较,AKT基因转染后,SGC-7901胃癌细胞中p-AKT^(Thr308)、Bcl-2等蛋白表达水平升高,FoxO1蛋白表达降低,与模型组比较,20、40 mol/L鱼藤素给药作用后能够降低p-AKT^(Thr308)、Bcl-2等蛋白的表达水平,降低Bcl-2 mRNA的表达水平,升高FoxO1及Bax的表达水平。结论:鱼藤素能够通过作用于AKT/FoxO1信号通路诱导胃癌细胞SGC-7901凋亡。

Objective:To explore the mechanism of deguelin(De)in inducing apoptosis of gastric cancer cells SGC-7901.Methods:The cytotoxicity of De at different concentrations(10,20,40,60,80,and 100 mol/L)for 24 and 48 h to SGC-7901 cells was investigated by cell counting kit-8(CCK-8)cell viability assay.SGC-7901 cells were divided into a control group and 20 and 40 mol/L De groups.After 24 h of administration,the expression levels of phosphor-protein kinase B(p-AKT)Thr308,forkhead box O1(FoxO1),and B-cell lymphoma-2(Bcl-2)proteins were determined by Western blot.AKT gene transfection was used to make the high expression of AKT in SGC-7901 cells.Then,20 and 40 mol/L of De were given for 24 h,and SGC-7901 cells that were not transfected with the AKT gene were used as the control group.Meanwhile,Western blot was used to determine the protein expression levels of p-AKTThr308,FoxO1,and Bcl-2,and real-time quantitative polymerase chain reaction(RT-qPCR)was used to determine the mRNA expression levels of FoxO1,Bcl-2,and Bcl-2 associated X protein(Bax).Results:Different concentrations of De showed certain cytotoxicity to gastric cancer cells SGC-7901.The 20 and 40 mol/L of De significantly decreased the protein expression levels of p-AKTThr308 and Bcl-2 in SGC-7901 cells,and increased the level of FoxO1 after treatment for 24 h.As compared with the control group,the protein expression levels of p-AKTThr308 and Bcl-2 in SGC-7901 cells were increased and FoxO1 was decreased after AKT gene transfection.As compared with the model group,the protein expression levels of p-AKTThr308 and Bcl-2 and the mRNA expression level of Bcl-2 were decreased,and the expression levels of FoxO1 and Bax were increased after treatment with 20 and 40 mol/L of De.Conclusion:De can induce the apoptosis of gastric cancer cells SGC-7901 by acting on the AKT/FoxO1 signaling pathway.