摘要
目的探究miR-27a在食管癌细胞中的表达及其对食管癌细胞增殖和上皮细胞间质转化(EMT)特性的影响。方法培养人正常食管鳞状上皮细胞株HEEC和食管癌细胞株ECA109、EC1、TE-4、CaEs-17。qRT-PCR检测各组细胞中miR-27a的表达。将细胞分为NC mimic组(miR-27a过表达阴性对照)、miR-27a mimic组(miR-27a过表达)、NC inhibitor组(miR-27a沉默阴性对照)、miR-27a inhibitor组(miR-27a沉默)。甲基噻唑基二苯基四氮唑(MTT)检测细胞增殖情况。Transwell检测细胞侵袭和迁移情况。qRT-PCR和Western blot分别检测细胞中EMT相关指标E-cadherin和vimentin mRNA和蛋白的表达情况。结果与正常食管鳞状上皮细胞株HEEC比较,食管癌细胞株ECA109、EC1、TE-4、CaEs-17中miR-27a的表达量均呈不同程度的升高,差异有统计学意义(F=28.790,P<0.05);其中ECA109细胞株升高最为显著,因此,选择这个细胞株作为后续实验。与NC mimic组细胞比较,miR-27a mimic组细胞增殖能力显著增强,迁移数目和侵袭数目均显著增多,vimentin mRNA和蛋白表达量显著升高,Ecadherin mRNA和蛋白表达量显著降低,差异均有统计学意义(P<0.05);与NC inhibitor组细胞比较,miR-27a inhibitor组细胞增殖能力显著减弱,迁移数目和侵袭数目均显著减少,vimentin mRNA和蛋白表达量显著降低,Ecadherin mRNA和蛋白表达量显著升高,差异均有统计学意义(P<0.05)。结论miR-27a在食管癌细胞中表达上调,且对食管癌细胞增殖具有促进作用,可通过调控EMT相关蛋白的水平,影响食管癌的EMT特性,进而促进食管癌细胞的侵袭和迁移。
Objective To investigate the expression of miR-27a in esophageal cancer cells and its effects on proliferation and epithelial mesenchymal transformation(EMT)characteristics.Methods Human normal esophageal squamous epithelial cell line HEEC and esophageal cancer cell lines ECA109,EC1,TE-4 and CaEs-17 were cultured.The expression of miR-27a was detected by q RT-PCR.The cells were divided into NC mimic group(miR-27a overexpression negative control),miR-27a mimic group(miR-27a overexpression),NC inhibitor group(miR-27a silencing negative control)and miR-27a inhibitor group(miR-27a silencing).Cell proliferation was detected by methylthiazolyldiphenyl-tetrazolium(MTT).Cell invasion and migration were detected by transwell test.q RT-PCR and Western blot were used to detect the expression level of E-cadherin and vimentin.Results Compared with normal esophageal squamous epithelial cell line HEEC,the expression of miR-27a in esophageal cancer cell lines ECA109,EC1,TE-4 and CaEs-17 increased in varying degrees,and the differences were statistically significant(F=28.790,P<0.05).And Eca109 cell line was the most significantly increased.Therefore,Eca109 cell line was selected as the follow-up experiment.Compared with NC mimic group,miR-27a mimic group significantly increased the proliferation,migration and invasion,vimentin mRNA and protein expression,and E-cadherin mRNA and protein expression(P<0.05).Compared with NC inhibitor group,the proliferation ability of miR-27a inhibitor group decreased significantly,the number of migration and invasion decreased significantly,the expression of vimentin mRNA and protein decreased significantly,and the expression of E-cadherin mRNA and protein increased significantly(P<0.05).Conclusions MiR-27a was up-regulated in esophageal cancer cells and could promote the proliferation of esophageal cancer.It could affect the EMT characteristics of esophageal cancer by regulating the level of EMT related proteins,so as to promote the invasion and migration of esophageal cancer cells.
作者
付民
尹先哲
尹星
常欢
周阳
邢艳丽
FU Min;YIN Xian⁃zhe;YIN Xing;CHANG Huan;ZHOU Yang;XING Yan⁃li(Oncology Department,Ward 1,the Second People's Hospital,Nanyang,Henan 473000,China)
出处
《热带医学杂志》
CAS
2022年第6期785-789,共5页
Journal of Tropical Medicine
基金
河南省医学科技攻关计划(2018020992)
关键词
miR-27a
食管癌
增殖
EMT特性
侵袭
迁移
MiR-27a
Esophageal cancer
Proliferation
EMT characteristics
Invasion
Migration
作者简介
付民(1989-),男,本科,主要从事肿瘤内科工作;通信作者:常欢,E⁃mail:yufuminggu@163.com
