miR-524-5p调控HEG1表达对食管癌细胞上皮间质转化的影响  

Effect of miR-524-5p on Epithelial-mesenchymal Transition in Esophageal Cancer Cells by Regulating HEG1 Expression

作　　者：王娅菲[1] 耿天祥[1] 陈林林[1] 李治国[1] 李世朋 WANG Yafei;GENG Tianxiang;CHEN Linlin;LI Zhiguo;LI Shipeng(Department of Gastroenterology,Jiaozuo People’s Hospital,Jiaozuo 454000,China;Liver Transplant Center,Beijing Friendship Hospital,Beijing 100050,China)

机构地区：[1]焦作市人民医院消化内科,焦作454000 [2]北京友谊医院肝移植中心,北京100050

出　　处：《肿瘤防治研究》2023年第11期1059-1065,共7页Cancer Research on Prevention and Treatment

基　　金：河南省医学科技项目科研课题(LHGJ20190371)。

摘　　要：目的探讨miR-524-5p调控HEG1表达对食管癌细胞增殖、上皮间质转化(EMT)的作用机制。方法qRT-PCR检测食管癌细胞和正常食管上皮细胞中miR-524-5p和HEG1mRNA表达水平。将KYSE30细胞分为miR-524-5pmimic组、miR-524-5pNC组、miR-524-5p mimic+pcDNA3.1组和miR-524-5p mimic+pc DNA3.1-HEG1组,另设置不转染的细胞为正常对照组(Control组)。CCK-8法检测KYSE30细胞增殖能力;Westernblot检测EMT相关蛋白、侵袭迁移相关蛋白和HEG1蛋白表达,划痕实验和Transwell实验检测KYSE30细胞迁移和侵袭能力,双荧光素酶报告基因检测miR-524-5p和HEG1靶向关系。结果miR-524-5p在四种食管癌细胞系TE-1、KYSE30、KYSE150、NEC中均低表达(P<0.05),选择表达水平最低的KYSE30细胞作为后续实验细胞。HEG1 mRNA在四种食管癌细胞中均高表达(P<0.05),并且GEPIA数据库显示HEG1在食管癌组织中高表达(P<0.05)。与miR-524-5pNC组相比,miR-524-5pmimic组KYSE30细胞增殖能力、克隆形成数、间质标志蛋白水平、迁移和侵袭能力显著降低,上皮标志蛋白E-cadherin水平显著上调(P<0.05);miR-524-5p mimic+pc DNA3.1-HEG1组可以明显逆转过表达mi R-524-5p对于KYSE30细胞增殖和上皮间质转化、侵袭转移的抑制作用(P<0.05)。miR-524-5pmimic组与WT-HEG1共转染组细胞荧光素酶活性显著低于miR-524-5p NC组与WT-HEG1共转染组(P<0.05)。结论miR-524-5p在食管癌细胞和组织中低表达,过表达miR-524-5p可以负向调节HEG1在食管癌细胞系KYSE30细胞中的表达,抑制KYSE30细胞的增殖、EMT进程和侵袭迁移能力。Objective To investigate the mechanism and the effect of miR-524-5p regulating HEG1 expression on the proliferation and epithelial-mesenchymal transition of esophageal cancer cells.Methods The expression levels of miR-524-5p and HEG1 mRNA in esophageal cancer cells and normal esophageal epithelial cells were detected by qRT-PCR.KYSE30 cells were divided into miR-524-5p mimic group,miR-524-5p NC group,miR-524-5p mimic+pcDNA3.1 group,and miR-524-5p mimic+pcDNA3.1-HEG1 group.Non-transfected cells were set as the normal control group(group Control).CCK-8 method was applied to detect the proliferation ability of KYSE30 cells.Western blot analysis was conducted to detect the expression of proteins related to EMT,invasion,and migration and the HEG1 protein.Scratch and Transwell assays were applied to detect the migration and invasion abilities of KYSE30 cells.A dual-luciferase reporter gene was used to examine the targeting relationship between miR-524-5p and HEG1.Results miR-524-5p was lowly expressed in four esophageal cancer cell lines,namely,TE-1,KYSE30,KYSE150,and NEC(P<0.05).KYSE30 cells with the lowest expression level were selected for subsequent experiments.HEG1 mRNA was highly expressed in four esophageal cancer cell lines(P<0.05).The GEPIA database showed that HEG1 was highly expressed in esophageal cancer tumor tissues(P<0.05).KYSE30 cells in the miR-524-5p mimic group had lower proliferation ability,colony formation number,mesenchymal marker protein expression,and migration and invasion abilities and upregulated epithelial marker protein E-cadherin level than cells in the miR-524-5p NC group(P<0.05).The miR-524-5p mimic+pcDNA3.1-HEG1 group significantly reversed the inhibitory effect of overexpression of miR-524-5p on the proliferation,epithelial–mesenchymal transformation,invasion,and metastasis of KYSE30 cells(P<0.05).The luciferase activity of cells in the miR-524-5p mimic and WT-HEG1 co-transfection groups was lower than that in the miR-524-5p NC and WT-HEG1 co-transfection groups(P<0.05).Conclusion m

关 键 词：miR-524-5p HEG1 食管癌 上皮间质转化 侵袭 

分 类 号：R735.1[医药卫生—肿瘤]