T-cadherin通过Wnt/ERK/PI3K-AKT信号通路在胃癌细胞自噬、细胞周期阻滞及上皮间质转化中的作用研究  

Effects of T-cadherin on autophagy,cell cycle arrest and epithelial-mesenchymal transition in gastric cancer cells via Wnt/ERK/PI3K-AKT signaling

作　　者：李立新[1] 蒲竞[2] 蔡翠芳[3] 李文详 LI Lixin;PU Jing;CAI Cuifang;LI Wenxiang(Department of Tumor Surgery,Lanzhou Second People s Hospital,Lanzhou 730046,China;Department of General Surgery,Lanzhou Second People s Hospital,Lanzhou 730046,China;Department of Neurourology,Lanzhou Second People s Hospital,Lanzhou 730046,China)

机构地区：[1]兰州市第二人民医院肿瘤外科,甘肃兰州730046 [2]兰州市第二人民医院普外科,甘肃兰州730046 [3]兰州市第二人民医院神经泌尿外科,甘肃兰州730046

出　　处：《中国肿瘤外科杂志》2020年第6期589-595,共7页Chinese Journal of Surgical Oncology

摘　　要：目的研究T-钙黏蛋白(T-cadherin,T-cad)通过Wnt/ERK/PI3K-AKT信号通路对胃癌细胞自噬、细胞周期阻滞及上皮向间质转化的影响。方法选取人胃癌细胞系AGS和人胃黏膜上皮细胞系GES-1为研究对象,将AGS细胞随机分为空白组、阴性对照(NC)组、T-cad组[和NC组、羟氯喹(HCQ)组、T-cad组、T-cad+HCQ组]。CCK-8法检测胃癌细胞AGS活力;克隆形成实验检测AGS细胞增殖能力;流式细胞术检测AGS细胞周期和凋亡情况;Western blotting检测AGS细胞中T-cadherin、自噬和Wnt/ERK/PI3K-AKT通路相关蛋白的表达。结果与GES-1组相比,空白组的T-cadherin表达显著降低(P<0.05);与NC组相比,T-cad组T-cadherin表达、细胞凋亡率、Bax、cleaved caspase 3和E-cadherin表达显著升高(P<0.05),细胞活力和细胞克隆数显著降低(P<0.05),G1期细胞比例显著升高,S期和G2/M期细胞比例显著降低(P<0.05),B淋巴细胞瘤-2(Bcl-2)、神经型钙黏蛋白(N-cadherin)、波形蛋白(Vimentin)、Wnt、β-catenin、p-ERK/ERK、PI3K和p-AKT/AKT蛋白表达显著降低(P<0.05);与NC组相比,HCQ组细胞Beclin1和LC3Ⅱ/LC3Ⅰ表达显著降低(P<0.05),细胞凋亡率无显著变化(P>0.05),T-cad组细胞Beclin1、LC3Ⅱ/LC3Ⅰ表达和细胞凋亡率显著升高(P<0.05),HCQ可逆转T-cad对AGS细胞自噬和凋亡的促进作用。结论T-cadherin可能通过Wnt/ERK/PI3K-AKT信号通路促进胃癌细胞自噬、细胞周期阻滞并抑制上皮间质转化。Objective To investigate the effects of T-cadherin on autophagy,cell cycle arrest and epithelial-to-mesenchymal transition of gastric cancer cells through Wnt/ERK/PI3K-AKT signaling pathway.Methods The human gastric cancer cell line AGS and human gastric mucosal epithelial cell line GES-1 were selected as the research subjects,and the AGS cells were randomly divided into divided into blank group,NC group and T-cad group(or NC group,HCQ group,T-cad group and T-cad+HCQ group),CCK-8 method was used to detect the viability of gastric cancer cell AGS;clone formation experiment was used to detect AGS cell proliferation;flow cytometry was used to detect AGS cell cycle and apoptosis;Western blotting was used to detect AGS cell T-cadherin,autophagy and Wnt/ERK/PI3K Expression of AKT pathway related proteins.Results Compared with GES-1 group,T-cadherin expression in AGS blank group was significantly reduced(P<0.05);compared with NC group,T-cadherin expression,apoptosis rate,Bax,cleaved caspase 3,E-cadherin expression was significantly increased(P<0.05),cell viability and cell clone number were significantly reduced(P<0.05),the proportion of cells in G1 phase was significantly increased,and the proportion of cells in S phase and G2/M phase was significantly reduced(P<0.05),Bcl-2,N-cadherin,Vimentin,Wnt,β-catenin,p-ERK/ERK,PI3K and p-AKT/AKT protein expression were significantly reduced(P<0.05);compared with the NC group,the expression of Beclin1 and LC3Ⅱ/LC3Ⅰin hydroxychloroquine(HCQ)group decreased significantly(P<0.05),the apoptosis rate did not change significantly(P>0.05),the expression of Beclin1,LC3Ⅱ/LC3Ⅰand apoptosis rate in T-cad group increased(P<0.05),HCQ can reverse the promotion effect of T-cad on autophagy and apoptosis of AGS cells.Conclusions T-cadherin may promote gastric cancer cell autophagy,cell cycle arrest and inhibit epithelial-mesenchymal transition through Wnt/ERK/PI3K-AKT signaling pathway.

关 键 词：T-CADHERIN 胃癌 自噬 细胞周期 上皮-间质转化 

分 类 号：R73[医药卫生—肿瘤]