miR-92b通过调控EZH2基因的表达抑制食管癌细胞Eca109的增殖和侵袭  

miR-92b suppresses growth and invasion of esophageal cancer Eca109 cells by regulating EZH2 gene expression

作　　者：刘飞[1] 孟令娇 刘世娜 谷丽娜[1] 李娟[1] 张建东 吴云艳[1] 桑梅香[1] 

机构地区：[1]河北医科大学第四医院科研中心,河北石家庄050011

出　　处：《中国肿瘤生物治疗杂志》2018年第2期118-124,共7页Chinese Journal of Cancer Biotherapy

基　　金：河北省杰出青年基金资助项目(No.2016206410)~~

摘　　要：目的:探讨食管癌(esophageal carcinoma,EC)细胞中miR-92b对组蛋白甲基转移酶zeste同源物增强子2(enhancer of zeste homolog 2,EZH2)基因表达的调控作用,以及对EC细胞增殖和侵袭能力的影响。方法:选取河北医科大学第四医院科研中心保存的2016年1月至2017年1月15例EC患者手术癌组织标本,通过生物信息学软件预测分析对EZH2可能调控的miRNAs,将预测的miRNAs mimic分别转染人EC细胞Eca109后,采用实时荧光定量PCR、Western blotting和双荧光素酶报告基因实验验证miRNAs对EZH2基因的靶向调控作用。同时将EZH2过表达质粒共转染至Eca109,然后采用CCK-8法、流式细胞术和Transwell细胞侵袭及迁移实验分别检测miRNAs和EZH2表达变化对EC细胞增殖、凋亡、侵袭和迁移的影响。结果:转染miR-92b的Eca109细胞中EZH2 mRNA与mimic-NC相比明显降低(P<0.01),转染miR-92b的Eca109细胞中EZH2蛋白表达水平明显低于转染mimic-NC组(0.525±0.052 vs 0.689±0.026,P<0.01)。生物信息学软件分析显示,miR-92b、let-7a及miR-25可与EZH2基因3’端非翻译区的结合位点相结合,但仅有miR-92b可以调控EZH2基因的表达,且miR-92b表达与EZH2 mRNA表达成负相关(P<0.01)。miR-92b mimic转染后EZH2 mRNA、蛋白及荧光素酶报告基因活性均明显下调(均P<0.01),对Eca109细胞凋亡无明显影响(P>0.05);miR-92b mimic转染能抑制ECa109细胞的增殖和侵袭及迁移能力(P<0.01)。而EC细胞转染EZH2过表达质粒后,miR-92b mimic对ECa109细胞增殖和侵袭及迁移能力的抑制作用明显减弱(P<0.01)。结论:miR-92b可抑制ECa109细胞的增殖和侵袭及迁移能力,其作用机制可能与靶向调控抑癌基因EZH2的表达有关。Objective： To investigate the regulating effects of miR-92b on the expression of EZH2（enhancer of zeste homolog 2） gene and the proliferation and invasion abilities of esophageal cancer（EC） cells. Methods： Fifteen cases of esophageal cancer tissues that preserved in the research center of the Fourth Hospital Affiliated to Heibei Medical University from January 2016 to January 2017 were selected for this study. The bioinformatics tool was used to predict the possible miRNAs that might target EZH2. The mimics of predicted miRNAs were transfected into human esophageal carcinoma cell lines Eca109, respectively. Then the regulation effect of miRNAs on EZH2 gene expression was validated by real-time PCR, Western blotting and dual luciferase reporter experiment. In the meanwhile,EZH2 over-expression plasmids were co-transfected into esophageal carcinoma Eca109 cells, and the effects of miRNAs and EZH2 expression changes on the proliferation, apoptosis, invasion and migration of esophageal carcinoma cells were detected by CCK-8 method, Flow Cytometry, Transwell Invasion and migration assay, respectively. Results： Bioinformatics analysis showed that miR-92b, let-7 a and miR-25 could combine with potential binding sites at 3＇-terminal non-translation region of EZH2 gene. Real-time PCR results showed that only miR-92b was able to regulate the expression of EZH2, and miR-92b was negatively correlated to EZH2 in esophageal cancer（P〈0.01）. Compared with mimic-NC, the expression of EZH2 mRNA, protein and luciferase activity in Eca109 cells after miR-92b mimic transfection was significantly down-regulated（both P〈0.01）. However, miR-92b mimic transfection had no effect on the apoptosis of Eca109 cells. Moreover, the proliferation, invasion and migration of Eca109 cells were significantly inhibited after transfection with miR-92b-mimic（P〈0.01）. In addition, after co-transfection with EZH2 over-expression plasmids, the effects of miR-92b-mimic on the proliferation, invasion and migration of Eca109

关 键 词：食管癌 miR-92b zeste同源物增强子2基因 增殖 侵袭 迁移 

分 类 号：R735.1[医药卫生—肿瘤] R730.4[医药卫生—临床医学]