基于PI3K/Akt/mTOR通路探讨川芎嗪诱导人胃癌细胞凋亡和自噬作用的机制研究  

Research on the Mechanism of TMP-induced Apoptosis and Autophagy in Human Gastric Cancer Cells Based on P13k/Akt/mTOR Pathway

作　　者：霍浩然[1] 秦瑞峰[1] 薛佳栋 袁增江[1] HUO Haoran;QIN Ruifeng;XUE Jiadong;YUAN Zengjiang(Handan Central Hospital,Handan 056001,China)

机构地区：[1]邯郸市中心医院,河北邯郸056001

出　　处：《西部中医药》2022年第10期12-16,共5页Western Journal of Traditional Chinese Medicine

基　　金：河北省医学科学研究重点课题计划(20181824)。

摘　　要：目的:研究川芎嗪(tetramethylpyrazine,TMP)对人胃癌SGC-7901细胞凋亡和自噬的影响并探索其分子机制。方法:分别以二甲基亚砜(dimethyl sulfoxide,DMSO)(空白对照组)、TMP(200、400、800μg/mL)和LY294002(5μg/mL)干预人胃癌SGC-7901细胞,48 h后通过MTT法检测细胞增殖抑制率,Annexin V-FITC/PI流式细胞术检测细胞凋亡水平,观察细胞自噬,Western blot法检测磷酸化磷酸肌醇3激酶(phosphorylated phosphatidylinositol 3-kinase,P-PI3K)、磷酸化蛋白激酶B(phosphorylated protein kinase B,p-Akt)、磷酸化哺乳动物雷帕霉素靶蛋白(phosphorylated mammalian target of rapamycin,p-mTOR)、活化型半胱氨酸蛋白酶9(cleaved cysteinyl aspartate specific proteinase 9,Cleaved Caspase-9)、活化型半胱氨酸蛋白酶3(cleaved cysteinyl aspartate specific proteinase 3,Cleaved Caspase-3)、微管相关蛋白1轻链3(microtubule-associated protein 1 light chain 3,LC3)蛋白表达,计算LC3-Ⅱ/LC3-Ⅰ比值。结果:TMP 200、400、800μg/mL或LY2940025μg/mL可明显提高细胞增殖抑制率、凋亡指数和自噬溶酶体数量。TMP 400、800μg/mL或PI3K特异性抑制剂LY2940025μg/mL可明显降低p-PI3K、p-Akt、p-mTOR相对表达量,提高Cleaved Caspase-9、Cleaved Caspase-3相对表达量和LC3-Ⅱ/LC3-Ⅰ比值,差异有统计学意义(P<0.05)。TMP 800μg/mL组对细胞增殖抑制,促其凋亡和自噬,p-PI3K、p-Akt、p-mTOR、Cleaved Cas-pase3表达及LC3-Ⅱ/LC3-Ⅰ比值的影响均明显优于LY2940025μg/mL组(P<0.05或P<0.01)。结论:TMP可能通过抑制PI3K/Akt/mTOR通路活化而诱导人胃癌细胞凋亡和自噬。Objective:To study the effects of TMP on the apoptosis and autophagy in human gastric cancer SGC-7901 cells and explore its molecular mechanism.Methods:DMSO(blank control group),TMP(200,400 and 800μg/mL)and LY294002(5μg/mL)were used to intervene human gastric cancer SGC-7901 cells,the inhibition rate of cell proliferation was measured via MTT method in 48 hours,the levels of cellular apoptosis was detected by using Annexin V-FITC/P1 flow cytometry(FCM),to observe cellular autophagy,the expressions of p-PI3K,p Akt,p-mTOR,Cleaved Caspase-9,Cleaved Caspase-3 and LC3 proteins were detected by using Western blot method,to calculate LC3-Ⅱ/LC3-Ⅰratio.Results:TMP in the concentrations of 200,400 and 800μg/mL or LY294002 in the concentration of 5μg/mL could notably raise the inhibition rate of cell proliferation,apoptotic index and the number of autophagic lysosomes.TMP in the concentrations of 400 and 800μg/mL or PI3K specific inhibitor LY294002 in the concentration of 5μg/mL could remarkably decrease the relative expressions of p-PI3K,p-Akt and p-mTOR,increase the relative expressions of Cleaved Caspase-9,Cleaved Caspase-3 and LC3-Ⅱ/LC3-Ⅰratio,and the difference had statistical meaning(P<0.05).TMP group of 800μg/mL demonstrated the more noticeable effects on the inhibition of cellular proliferation,promoting the apopotosis and autophagy of the cells,the expressions of p-PI3K,p-Akt,p-mTOR,Cleaved Caspase-3 and LC3-Ⅱ/LC3-Ⅰratio compared with the effects of LY294002 group of 5μg/mL(P<0.05 or P<0.01).Conclusion:TMP could induce the apoptosis and autophagy of human gastric cancer cells possibly through inhibiting the activation of PI3K/Akt/mTOR pathway.

关 键 词：胃癌 川芎嗪 PI3K/AKT/MTOR 凋亡 自噬 

分 类 号：R735[医药卫生—肿瘤]