MicroRNA-133a-5p调控c-met表达对胆管癌细胞迁移和侵袭的影响  

The Effect of MicroRNA-133a-5p on the Migration and Invasion of Cholangiocarcinoma Cells Through Regulatingc-met

作　　者：杨波[1] 刘小方[1] YANG Bo;LIU Xiao-fang(Department of Hepatobiliary-Pancreatic Surgery,Affiliated YantaiYuhuangding Hospital,Qingdao University,Yantai(264000),China)

机构地区：[1]青岛大学附属烟台毓璜顶医院肝胆胰外科,烟台264000

出　　处：《中国中西医结合外科杂志》2020年第3期429-434,共6页Chinese Journal of Surgery of Integrated Traditional and Western Medicine

基　　金：北京医卫健康公益基金会医学科学研究基金资助项目(B17134)。

摘　　要：目的:探讨MicroRNA-133(miR-133)在胆管癌细胞系中的表达,研究miR-133基因表达在胆管癌细胞迁移和侵袭过程中的作用。方法:利用实时荧光定量PCR(RT-PCR)技术测定分析miR-133及其不同成熟体(miR-133b、miR-133a-3p、miR-133a-5p)在胆管癌QBC939细胞系中的表达。将胆管癌QBC939细胞系经转染后分为三组:自然生长组(空白对照组),细胞转染无关序列组(阴性对照组)以及细胞转染Anti-miR-133a-5p组(干扰组),利用RT-PCR检测各组胆管癌QBC939细胞系中miR-133a-5p、c-met的表达。然后利用Transwell小室法分别检测分析干扰miR-133a-5p基因表达对胆管癌QBC939细胞系迁移和侵袭能力的影响。结果:RT-PCR检测miR-133不同成熟体miR-133b(0.74±0.07)、miR-133a-3p(0.99±0.12)、miR-133a-5p(1.00±0.06)中的表达,其中miR-133a-5p在胆管癌QBC939细胞系中表达量最高(P<0.05)。RT-PCR实验证实干扰组QBC939细胞系中miR-133a-5p(0.70±0.08)表达显著低于对照组(1.43±0.34);干扰组QBC939细胞系中c-met(0.09±0.02)表达也显著低于对照组(1.01±0.07),差异有统计学意义(P<0.01)。Transwell小室实验证实干扰组中胆管癌QBC939细胞系迁移(69.5±8.3)和侵袭(19.3±3.3)能力明显低于对照组,差异均有统计学意义(P<0.01)。结论:MicroRNA-133a-5p的低表达可以明显减弱c-met的表达,同时可以抑制胆管癌QBC939细胞系的迁移和侵袭能力,miR-133a-5p有可能成为胆管癌基因表达调控的新靶点。Objective To investigate the expression of MicroRNA-133(miR-133)in cholangiocarcinoma cells and to study the role of miR-133 gene expression in the migration and invasion of cholangiocarcinoma cells.Methods Real-time PCR(RT-PCR)was used to analyze the expression of miR-133 and its different matures(miR-133b、miR-133a-3p、miR-133a-5p)in cholangiocarcinoma cells QBC939.QBC939 cells were chosen and divided into three groups:natural growth(blank control group),cellgroups transfection with negative sequence(negative control group)and cell groups transfection with anti-miR-133a-5p(interference group).The expression of miR-133a-5pand c-met in the QBC939 cells of each group were measured by RT-PCR.The effect of miR-133a-5p on the migration and invasion of QBC939 cells was analyzed by Transwell assay.Results The expression of miR-133b(0.74±0.07),miR-133a-3p(0.99±0.12)and miR-133a-5p(1.00±0.06)in miR-133 was detected by RT-PCR,miR-133a-5p had the highest expression in cholangiocarcinoma cells QBC939(P<0.05).RT-PCR experiments confirmed that the expression of miR-133a-5p(0.70±0.08)in the interference group QBC939 cells was significantly lower than that in the negative control group(1.43±0.34,P<0.05)and the expression of c-met(0.09±0.02)in the interference group QBC939 cells was significantly lower than that in the blank control group(1.01±0.07,P<0.01).Transwell experiments confirmed that the migration(69.5±8.3)and invasion(19.3±3.3)abilities of QBC939 cells in the interference group were significantly lower than that of the negative and blank control groups(both P<0.01).Conclusion The low expression of microRNA-133a-5p can significantly attenuate the expression of c-met and inhibit the migration and invasion of cholangiocarcinoma cells QBC939.The miR-133a-5pmaybecome a new target for the regulation of cholangiocarcinoma gene expression.

关 键 词：微小RNA-133 胆管癌 C-MET 迁移 侵袭 

分 类 号：R657.15[医药卫生—外科学]