miR-133b通过介导TGF-βR1表达下调对食管癌细胞侵袭、迁移的抑制作用  

MiR-133b inhibits the invasion and migration of esophageal cancer cells by mediating the down-regulation of TGF-βR1 expression

作　　者：米卫国[1] 张伟[1] 刘建军 张昭鹏 杨帆[1] MI Weiguo;ZHANG Wei;LIU Jianjun;ZHANG Zhaopeng;YANG Fan(Department of Thoracic Surgery,Hanzhong Municipal Central Hospital,Hanzhong,Shaanxi 723000,China)

机构地区：[1]陕西省汉中市中心医院胸外科,陕西汉中723000

出　　处：《检验医学与临床》2021年第8期1084-1088,1092,共6页Laboratory Medicine and Clinic

摘　　要：目的探讨miR-133b通过介导转化生长因子-β受体1(TGF-βR1)表达下调,抑制食管癌细胞侵袭、迁移的作用及机制。方法采用实时荧光定量PCR检测人正常食管上皮细胞和人食管癌OE19、ECA109细胞中miR-133b的表达;构建miR-133b过表达的人食管癌OE19、ECA109细胞;Transwell小室实验观察不同miR-133b表达水平对食管癌OE19、ECA109细胞迁移和侵袭能力的影响;蛋白免疫印迹法检测不同miR-133b表达水平对食管癌OE19、ECA109细胞中TGF-βR1、SMAD3、p-SMAD3、E-cadherin、N-cadherin蛋白水平的影响;双荧光素酶报告基因试验检测miR-133b对TGF-βR1的靶向调控作用。结果成功构建miR-133b过表达的食管癌OE19和ECA109细胞;miR-133b过表达的食管癌OE19和ECA109细胞的侵袭和迁移能力显著降低(P<0.01);miR-133b过表达的食管癌OE19和ECA109细胞中TGF-βR1、p-SMAD3、N-cadherin蛋白水平明显降低(P<0.01),E-cadherin蛋白水平明显升高(P<0.01)。双荧光素酶报告基因试验检测发现miR-133b可靶向调控TGF-βR1的表达。结论 miR-133b通过介导TGF-βR1表达下调,抑制TGF-βR1/SMAD3信号通路活化,抑制人食管癌OE19和ECA109细胞上皮间质转化的发生,抑制食管癌细胞的侵袭和迁移能力。Objective To investigate the role and mechanism of miR-133b in inhibiting the invasion and migration of esophageal cancer cells by mediating the down-regulation of transforming growth factor-βreceptor 1(TGF-βR1)expression.Methods Real-time fluorescent quantitative PCR was used to detect the expression level of miR-133b in human normal esophageal epithelial cells and human esophageal cancer OE19 and ECA109 cells.Human esophageal cancer OE19 and ECA109 cells overexpressed miR-133b were constructed.Transwell chamber experiment was used to observe the effect of miR-133b expression on the migration and invasion of OE19 and ECA109 cells.Western blotting was used to detect the effects of different miR-133b expression levels on TGF-βR1,SMAD3,p-SMAD3,E-cadherin and N-cadherin protein expression levels in esophageal cancer OE19 and ECA109 cells.The dual luciferase reporter gene test was used to detect the targeted regulation effect of miR-133b on TGF-βR1.Results Esophageal epithelial cancer OE19 and ECA109 cells with miR-133b overexpression were successfully constructed.The invasion and migration ability of OE19 and ECA109 cells with miR-133b overexpression were significantly decreased(P<0.01).The protein levels of TGF-βR1,p-SMAD3 and N-cadherin in OE19 and ECA109 cells were significantly decreased(P<0.01),and E-cadherin protein was significantly increased(P<0.01).The dual luciferase reporter gene test showed that miR-133b could target and regulate the expression of TGF-βR1.Conclusion By mediating the down-regulation of TGF-βR1 expression,miR-133b can inhibit the activation of TGF-βR1/SMAD3 signaling pathway,inhibit epithelial mesenchymal transition occurrence of OE19 and ECA109 cells,and inhibit the invasion and migration abilities of esophageal cancer cells.

关 键 词：食管癌 miR-133b 转化生长因子-β受体1 侵袭 迁移 

分 类 号：R735.1[医药卫生—肿瘤]