HMGA2通过Wnt/β-catenin信号通路促进结直肠癌上皮间质转化的研究  

HMGA2 promotes colorectal cancer epithelial-mesenchymal transition through Wnt/β-catenin signaling pathway

作　　者：张文俊[1] 王明祥 宋沁哲 张福杰[1] 于登峰[1] 罗福文[2] ZHANG Wen-jun;WANG Ming-xiang;SONG Qin-zhe;ZHANG Fu-jie;YU Deng-feng;LUO Fu-wen(Anorectal department,,Dalian University Affiliated Xinhua Hospital Anorectal,DaLian 116622,China;Acute Abdominal Surgery,The Second Hospital of Dalian Medical University,DaLian 116027,China)

机构地区：[1]大连大学附属新华医院肛肠外科,辽宁大连116021 [2]大连医科大学附属第二医院急腹症外科,辽宁大连116027

出　　处：《中国现代普通外科进展》2023年第4期253-257,共5页Chinese Journal of Current Advances in General Surgery

基　　金：大连青年科技之星(基金编号:2021RQ030);大连市医学科学研究计划(基金编号:2111039)。

摘　　要：目的:研究高迁移率蛋白A2(HMGA2)通过Wnt/β-catenin信号通路促进结直肠癌上皮间质转化(EMT)的影响。方法:采用Real-time PCR法和Western blot法测定正常人结直肠上皮细胞和人结直肠癌HCT116细胞中HMGA2表达水平;将HCT166细胞转染GV144-HMGA2质粒过表达HMGA2后,测定过表达HMGA2对HCT166细胞活力、凋亡及迁移的影响;转染GV144-HMGA2质粒6 h后,加入重组转化生长因子(TGF)-β1蛋白继续培养48 h,测定EMT标志物和细胞周期蛋白(cyclin)D1表达水平。HCT166细胞中分别加入Wnt/β-catenin信号通路的激活剂(LiCl)和抑制剂(XAV93920),并加入TGF-β1和过表达的HMGA2,检测EMT标志物和cyclin D1表达水平。结果:HCT166细胞中HMGA2表达水平显著高于FHC细胞(P<0.05);过表达HMGA2可以显著增加HCT166的细胞活力和迁移,降低细胞凋亡(P<0.05);HMGA2可以显著降低E-caderin表达水平,降低TGF-β1诱导的cyclin D1、c-Myc、Snail、vimentin和β-catenin的表达水平(P<0.05)。加入Wnt/β-catenin信号通路激活剂后,HMGA2对EMT标志物的影响显著增加,而加入Wnt/β-catenin信号通路抑制剂后,HMGA2对EMT标志物的影响显著降低(P<0.05)。结论:过表达HMGA2可以通过Wnt/β-catenin信号通路促进HCT166细胞的生长和迁移,降低细胞凋亡,促进EMT的发生。Objective:To study the effect of HMGA2 on promoting the epithelial-mesenchymal transition(EMT)of colorectal cancer through the Wnt/β-catenin signaling pathway.Methods:Real-time PCR and Western blot were used to determine the expression level of HMGA2 in normal human colorectal epithelial cells and human colorectal cancer HCT116 cells,After transfecting HCT166 cells into GV144-HMGA2 plasmid overexpressing HMGA2,determine the effect of overexpression of HMGA2 on the viability,apoptosis and migration of HCT166 cells;Six hours after transfection of GV144-HMGA2 plasmid,recombinant transforming growth factor(TGF)-β1 protein was added and cultured for 48 hours,and then the expression levels of EMT markers and cyclin D1 were measured.Then,the Wnt/β-catenin signaling pathway activator(LiCl)and inhibitor(XAV93920)were added to HCT166 cells,and TGF-β1 and overexpressed HMGA2 were added to detect the expression levels of EMT markers and cyclin D1.Results:The expression level of HMGA2 in HCT166 cells was significantly higher than that in FHC cells(P<0.05),Overexpression of HMGA2 can significantly increase the cell viability and migration of HCT166,and reduce cell apoptosis(P<0.05),HMGA2 can significantly reduce the expression level of E-caderin,and reduce the expression levels of cyclin D1,c-Myc,Snail,vimentin andβ-catenin induced by TGF-β1(P<0.05).After adding the Wnt/β-catenin signaling pathway activator,the effect of HMGA2 on EMT markers increased significantly,and after adding the Wnt/β-catenin signaling pathway inhibitor,the effect of HMGA2 on EMT markers was significantly reduced(P<0.05).Conclusion:Overexpression of HMGA2 can promote the growth and migration of HCT166 cells through the Wnt/β-catenin signaling pathway,reduce cell apoptosis,and promote the occurrence of EMT.

关 键 词：结直肠肿瘤 高迁移率蛋白A2 WNT/Β-CATENIN信号通路 上皮间质转化 

分 类 号：R735.35[医药卫生—肿瘤]