MicroRNA-145增强非小细胞肺癌细胞对吉非替尼敏感性及其机制探讨  

The effect and mechanism of microRNA-145 enhances the gefitinib sensitivity in non-small cell lung cancer cells

作　　者：邓建松 魏柏 Deng Jiansong;Wei Bai(Department of Oncology,Liyuan Hospital,the Affiliated Hospital of Tongji Medical University,Huazhong University of Science and Technology,Hubei Wuhan 430077,China)

机构地区：[1]华中科技大学同济医学院附属梨园医院肿瘤科,湖北武汉430077

出　　处：《现代肿瘤医学》2020年第20期3511-3516,共6页Journal of Modern Oncology

摘　　要：目的:探讨微小RNA-145(microRNA-145,miR-145)对非小细胞肺癌细胞系吉非替尼耐药的作用及其可能的潜在机制。方法:实时荧光定量PCR(Q-PCR)方法检测正常细胞及非小细胞肺癌细胞中miR-145的表达差异;不同时间5μmol/L吉非替尼干预肺癌细胞SPC-A-1和A549后,Q-PCR检测miR-145表达变化;miR-145 mimics和miR-145 inhibitors分别转染SPC-A-1和A549肺癌细胞后,CCK8检测细胞活力变化;双荧光素酶报告系统检测miR-145与ADAM19的靶向结合;miR-145 mimics转染SPC-A-1和A549肺癌细胞,Western blot检测ADAM19蛋白表达;Western blot检测5μmol/L吉非替尼干预后,SPC-A-1和A549肺癌细胞中ADAM19蛋白的表达;mi R-145 mimics转染SPC-A-1和A549肺癌细胞,再用5μmol/L吉非替尼干预,Western blot检测ADAM19蛋白的表达;采用si RNA抑制ADAM19表达,再用5μmol/L吉非替尼干预,CCK8检测细胞活力;采用BALB/C雌性裸鼠皮下接种肿瘤细胞的方法,观察上述效应。结果:QPCR结果显示,与正常肺上皮细胞系BEAS-2B相比较,肺癌细胞SPC-A-1和A549中mi R-145表达明显降低;5μmol/L吉非替尼干预SPC-A-1和A549细胞后,mi R-145表达显著上调;转染mi R-145 mimics后,CCK8结果显示两种细胞细胞活力下降;双荧光素酶报告系统结果显示,mi R-145靶向结合ADAM19基因的3’-UTR区;Western blot结果显示,5μmol/L吉非替尼诱导SPC-A-1和A549细胞后,两种细胞中ADAM19蛋白表达均降低;mi R-145 mimics转染SPC-A-1和A549细胞,之后给予5μmol/L吉非替尼治疗,ADAM19蛋白表达下调;si RNA抑制SPC-A-1和A549细胞中ADAM19表达,再给予5μmol/L吉非替尼治疗,CCK8结果显示,两种细胞的细胞活力显著降低;过表达BALB/C雌性裸鼠的mi R-145后,显著抑制皮下肿瘤生长,并且增强吉非替尼的抗肿瘤效果。结论:mi R-145显著增强肺癌细胞SPC-A-1和A549对吉非替尼的敏感性,其机制可能是通过靶向结合AMDM19基因的3’-UTR区域,从而抑制其表达。mi R-145有可能成为治疗非小细胞肺癌Objective:To investigate the association between miR-145 and gefitinib resistance in non-small cell lung cancer(NSCLC)cell lines and potential mechanism.Methods:Real-time fluorescence quantitative PCR(Q-PCR)was used to detect the difference of miR-145 level in normal cells and lung cancer cells,and the miR-145 level in SPC-A-1 and A549 lung cancer cells treated with gefitinib at different times.After SPC-A-1 and A549 cells were transfected with miR-145 mimics and miR-145 inhibitors,CCK8 detected the cell viability.The double-luciferase reporter system detected the action site of miR-145.Western blot detected the protein expression of ADAM19 gene in SPC-A-1 and A549 cells after 5μmol/L gefitinib treatment.After transfection with miR-145 mimics and then treatment with 5μmol/L gefitinib,Western blot detected the ADAM19 protein expression in SPC-A-1 and A549 cell.SPC-A-1 and A549 cells were firstly administrated with siRNA-ADAM19,then treated with 5μmol/L gefitinib.CCK8 detected the cell activity.The above effect was observed by subcutaneous inoculation of tumor cells in BALB/C female nude mice.Results:Q-PCR showed that compared with normal lung epithelial cell line BEAS-2 B,the level of miR-145 in human lung cancer cells SPC-A-1 and A549 cells were significantly decreased.miR-145 expression was significantly increased after 5μmol/L gefitinib intervention in SPC-A-1 and A549 cells.After transfection with miR-145 mimics,the cell viability of the two types of cells were decreased.The double-luciferase reporter system results showed that miR-145 targeted the 3’-UTR region of ADAM19 gene.After 5μmol/L gefitinib incubation with SPC-A-1 and A549 cells,the protein levels of ADAM19 in both cells were decreased.After transfection with miR-145 mimics,the SPC-A-1 and A549 cells were then treated with 5μmol/L gefitinib.The ADAM19 protein expression in two type cells was significantly down-regulated.siRNA inhibited the ADAM19 expression in SPC-A-1 and A549 cells,and then treated with 5μmol/L gefitinib.CCK8 showed that th

关 键 词：非小细胞肺癌 耐药 microRNA-145 

分 类 号：R734.2[医药卫生—肿瘤]