miR-34b-5p通过Gab1调节mTOR通路抑制子宫内膜癌细胞的增殖迁移  

Mechanism of miR-34b-5p Targeting Gab1 and Regulating mTOR Pathway to Inhibit the Proliferation and Migration of EndometrialCancer Cells

作　　者：田雪敏 石璐璐 董辉[2] 杨彩虹 TIAN Xuemin;SHI Lulu;DONG Hui;YANG Caihong(School of Clinical Medical Sciences,Ningxia Medical University,Yinchuan 750004,China;Institute of Medical Sciences,General Hospital of Ningxia Medical University,Yinchuan 750004,China;Department of Obstetrics and Gynecology,General Hospital of Ningxia Medical University,Yinchuan 750004,China)

机构地区：[1]宁夏医科大学临床医学院,银川750004 [2]宁夏医科大学总医院医学科学研究院,银川750004 [3]宁夏医科大学总医院心脑血管病医院妇科,银川750004

出　　处：《宁夏医科大学学报》2022年第11期1094-1099,1104,共7页Journal of Ningxia Medical University

基　　金：国家自然科学基金项目(81960471)。

摘　　要：目的探讨miR-34b-5p对子宫内膜癌(endometrial cancer,EC)细胞增殖和迁移的作用及机制。方法qRT-PCR方法检测miR-34b-5p、Gab1在EC及癌旁组织中的表达水平,双荧光素酶基因报告(293T细胞)实验验证miR-34b-5p和Gab1的靶向关系,建立miR-34b-5p慢病毒过表达的人子宫内膜腺癌细胞AN3CA细胞模型,qRT-PCR方法检测EC和癌旁组织中miR-34b-5p的表达水平,CCK-8、划痕实验检测AN3CA细胞增殖、迁移能力,Western blot方法检测Gab1、p-mTOR、p-p70s6k、p-4EBP的表达水平。结果EC组织中的基因表达结果显示,miR-34b-5p呈现低表达,Gab1呈现高表达(P均<0.05);qRT-PCR结果显示,成功构建miR-34b-5p过表达的AN3CA细胞株;双荧光素酶实验结果显示,miR-34b-5p具有靶向Gab1 mRNA的结合能力;CCK-8结果显示miR-34b-5p过表达抑制AN3CA细胞的增殖(P<0.05);划痕实验显示miR-34b-5p过表达降低AN3CA细胞的迁移能力(P<0.05);Western blot结果显示miR-34b-5p的过表达抑制Gab1、p-mTOR、p-p70s6k、p-4EBP蛋白的表达水平(P均<0.05)。结论miR-34b-5p过表达可抑制AN3CA的增殖和迁移,与Gab1介导的p-mTOR通路下调有关。Objective To explore the effect and mechanism of miR-34b-5p regulating Gab1-mediated mTOR pathway on the proliferation and migration of endometrial cancer cells.Methods The qRT-PCR method was used to detect the expression levels of miR-34b-5p and Gab1 in EC and paracancerous tissues,the targeting relationship between miR-34b-5p and Gab1 was verified by the double luciferase gene reporting(293T cells)experiment,and a model of miR-34b-5p lentivirus overexpression of human endometrial adenocarcinoma cells AN3CA cells was established.qRT-PCR was used to detect the expression level of miR-34b-5p in EC and adjacent tissues,CCK-8 and scratch test were used to detect the proliferation and migration ability of AN3CA cells,and Western blot was used to detect the expression level of Gab1,p-mTOR,p-p70s6k and p-4EBP.Results The gene expression results showed that miR-34b-5p showed low expression,while Gab1 showed high expression(P<0.05).RT-qPCR results showed that AN3CA cell line overexpressed miR-34b-5p was successfully constructed.The results of double luciferase assay showed that miR-34b-5p had the binding ability to target Gab1 mRNA.CCK-8 results showed that miR-34b-5p overexpression inhibited the proliferation of AN3CA cells(P<0.05).Wound Healing assay showed that miR-34b-5p overexpression decreased the migration ability of AN3CA cells(P<0.05).Western blot results showed that miR-34b-5p overexpression inhibited the expression levels of Gab1,p-mTOR,p-p70s6k and p-4EBP proteins(P all<0.05).Conclusion miR-34b-5p inhibits Gab1 expression,down-regulates p-mTOR pathway,and inhibits the proliferation and migration of AN3CA in endometrial cancer cells.

关 键 词：miR-34b-5p Grb2相关结合蛋白 子宫内膜癌 增殖 

分 类 号：R737.33[医药卫生—肿瘤]