摘要
目的观察hsa-miR-124-3p.1通过靶向肿瘤坏死因子受体相关因子6(TRAF6)抑制转化生长因子β1(TGF-β1)对人胃癌细胞上皮间质转化、迁移和侵袭的影响。方法收集胃癌组织和癌旁正常组织各43例,培养人胃黏膜上皮细胞GES-1及胃癌细胞NCI-N87、MGC-803、BGC-823、SGC-7901、MKN-45,采用荧光定量PCR技术和蛋白印迹法检测组织和细胞中miR-124-3p.1、TRAF6的表达情况;采用双荧光素酶报告基因系统实验验证miR-124-3p.1和TRAF6的靶向关系;构建过表达miR-124-3p.1、TRAF6的SGC-7901细胞株,以TGF-β1诱导细胞,采用Transwell小室实验、划痕实验评估细胞侵袭和迁移能力。结果与癌旁正常组织相比,胃癌组织中miR-124-3p.1表达下调,TRAF6的mRNA和蛋白表达上调(P<0.05);与正常胃黏膜培养细胞比较,胃癌细胞中有类似变化。与对照组比较,TGF-β1组细胞中E-cadherin表达下调,N-cadherin和Vimentin表达上调,细胞侵袭率和迁移率增加(P<0.05);与TGF-β1组比较,转染miR-124-3p.1 mimic细胞中E-cadherin表达上调,N-cadherin和Vimentin表达下调,细胞侵袭率和迁移率降低(P<0.05)。与miR-124-3p.1 mimic组比较,TGF-β1+mimic+TRAF6组细胞侵袭率、迁移率增加,TRAF6、N-cadherin和Vimentin表达上调,E-cadherin表达下调(P<0.05)。结论 hsa-miR-124-3p.1在胃癌中呈低表达,过表达miR-124-3p.1可抑制TGF-β1诱导的上皮间质转化、细胞侵袭和迁移,其作用机制可能与靶向下调TRAF6表达有关。
Objective To observe the effects of hsa-miR-124-3p. 1 in inhibiting epithelial-mesenchymal transition(EMT), migration and invasion of human gastric cancer cells induced by transforming growth factor β1(TGF-β1)by targeting tumor necrosis factor receptor-associated factor 6(TRAF6). Methods A total of 43 gastric cancer tissues and 43 normal para-carcinoma tissues were collected. The human gastric mucosal epithelial cells GES-1 and gastric cancer cells(NCI-N87, MGC-803, BGC-823, SGC-7901, and MKN-45) were cultured. The expressions of miR-124-3p. 1 and TRAF6 in tissues and cells were detected by fluorescent quantitative PCR and Western blotting. The targeted relationship between miR-124-3p.1 and TRAF6 was verified by dual-luciferase reporter gene system assay. SGC-7901 cell lines with miR-124-3p. 1 and TRAF6 overexpression were constructed. The cells were induced by TGF-β1. The invasion and migration abilities of the cells were evaluated by Transwell chamber assay and scratch test. Results Compared with normal para-carcinoma tissues and normal gastric mucosal cells, the expression of miR-124-3p. 1 was downregulated,while the expressions of TRAF6 mRNA and protein were upregulated in gastric cancer tissues and cells(P<0. 05).Compared with control group, expression of E-cadherin in cells was downregulated, expressions of N-cadherin and Vimentin were upregulated,invasion and migration rates of cells were increased in TGF-β1 group(P<0. 05). Compared with TGF-β1 group,after cells were transfected with miR-124-3p. 1 mimic,the expression of E-cadherin was upregulated,the expressions of N-cadherin and Vimentin were down-regulated,and invasion and migration rates of cells were decreased(P<0. 05). Compared with miR-124-3p.1 mimic group,invasion and migration rates of cells were increased in TGF-β1+mimic+TRAF6 group, expressions of TRAF6, N-cadherin and Vimentin were up-regulated, and the expression of E-cadherin was down-regulated(P<0. 05). Conclusion hsa-miR-124-3p. 1 is lowly expressed in gastric cancer.Overexpression of
作者
陶正贵
杜静虎
田葵
王东华
胡凤琪
陈满宇
TAO Zhenggui;DU Jinghu;TIAN Kui;WANG Donghua;HU Fengqi;CHEN Manyu(Department of General Surgery,Xiangyang Central Hospital,Affiliated Hospital of Hubei University of Arts and Sciences,Xiangyang 441000;Department of Nephrology,Xiangyang Central Hospital,Xiangyang 441000,China)
出处
《西安交通大学学报（医学版）》
CAS
CSCD
北大核心
2021年第6期843-849,共7页
Journal of Xi’an Jiaotong University（Medical Sciences）
基金
湖北卫生计生委青年科技人才项目(No.WJ2015Q038)。
