miR-497-5p靶向VEGFA抑制子宫内膜癌Ishikawa细胞侵袭、迁移和上皮间质转化的实验研究  

MiR-497-5p Targets VEGFA to Inhibit Invasion,Migration and Epithelial-Mesenchymal Transition of Endometrial Cancer Ishikawa Cells

作　　者：申利 刘佳[1] 林星光 SHEN Li;LIU Jia;LIN Xing-guang(Department of Obstetrics and Gynecology of the Affiliated Liyuan Hospital,Huazhong University of Science and Technology,Wuhan 430077,China;Department of Obstetrics and Gynecology of the Affiliated Tongji Hospital,Tongji Medical College of Huazhong University of Science and Technology,Wuhan 430030,China)

机构地区：[1]华中科技大学同济医学院附属梨园医院妇产科,武汉430077 [2]华中科技大学同济医学院附属同济医院妇产科,武汉430030

出　　处：《南昌大学学报（医学版）》2021年第1期1-5,38,共6页Journal of Nanchang University：Medical Sciences

基　　金：国家自然科学基金(81701476);湖北省卫生健康委员会面上项目(WJ2019M138)。

摘　　要：目的探讨miR-497-5p对子宫内膜癌Ishikawa细胞侵袭、迁移和上皮间质转化(EMT)的影响及其机制。方法将体外培养的Ishikawa细胞分为miR-NC组、miR-497-5p组、pLV-VEGFA组和miR-497-5p+VEGFA组,采用Real time PCR检测miR-497-5p和血管内皮生长因子A(VEGFA)mRNA的表达,双荧光素酶报告基因实验检测miR-497-5p和VEGFA的靶向关系,Western blot检测VEGFA蛋白和EMT相关蛋白神经型钙黏蛋白(N-cadherin)、波形蛋白(Vimentin)和E-钙黏蛋白(E-cadherin)的表达,Transwell小室实验检测Ishikawa细胞的侵袭和迁移。结果转染miR-497-5p mimics可升高Ishikawa细胞中miR-497-5p表达,降低VEGFA mRNA表达(P<0.05),VEGFA是miR-497-5p的靶基因。与miR-NC组比较,miR-497-5p组细胞中VEGFA、Vimentin和E-cadherin蛋白的表达水平、侵袭细胞数和迁移细胞数均明显降低,而N-cadherin蛋白表达升高(P<0.05);与pLV-VEGFA组比较,miR-497-5p+VEGFA组细胞中VEGFA、Vimentin和E-cadherin蛋白的表达水平以及侵袭、迁移细胞数量均明显降低,而N-cadherin蛋白表达明显升高(P<0.05)。结论miR-497-5p可靶向VEGFA抑制子宫内膜癌Ishikawa细胞的侵袭、迁移和上皮间质转化。Objective To investigate the effects of miR-497-5p on invasion,migration and epithelial-mesenchymal transformation(EMT)of endometrial cancer Ishikawa cells and its mechanisms.Methods Ishikawa cells cultured in vitro were divided into miR-NC group,miR-497-5p group,pLV-VEGFA group and miR-497-5p+VEGFA group.The expression of miR-497-5p and vascular endothelial growth factor A(VEGFA)was detected by real-time PCR.The targeting relationship between miR-497-5p and VEGFA was checked by double luciferase reporter gene assay.The levels of VEGFA protein and EMT-related proteins N-cadherin,Vimentin and E-cadherin were measured by Western blot.The invasion and migration of Ishikawa cells were determined by transwell assay.Results Transfection with miR-497-5p mimics increased miR-497-5p expression and decreased VEGFA mRNA expression in Ishikawa cells.VEGFA was a target gene of miR-497-5p.Compared with microRNA-NC group,the expression of VEGFA,Vimentin and E-cadherin proteins and the number of invasive and migrated cells decreased,but the expression of N-cadherin protein increased in miR-497-5p group(P<0.05).Compared with pLV-VEGFA group,the expression of VEGFA,Vimentin and E-cadherin proteins and the number of invasive and migrated cells decreased,while the expression of N-cadherin protein increased in miR-497-5p+VEGFA group(P<0.05).Conclusion miR-497-5p can target VEGFA to inhibit the invasion,migration and EMT of endometrial cancer Ishikawa cells.

关 键 词：miR-497-5p 子宫内膜癌 细胞侵袭 迁移 上皮间质转化 血管内皮生长因子A 

分 类 号：R737.33[医药卫生—肿瘤]