miRNA-140介导Smad3对非小细胞肺癌的生物学作用  

Biological effects of Smad3 mediated by microRNA-140 on non-small cell lung cancer cells

作　　者：尹端端[1] 杨雁鸿[1] 周文文[1] 田力 王小玉 孙艳[1] 赵子舒 王海霞 刘岩[2] YIN Duan-duan;YANG Yan-hong;ZHOU Wen-wen;TIAN Li;WANG Xiao-yu;SUN Yan;ZHAO Zi-shu;WANG Hai-xia;LIU Yan(Department of Oncology,Qinhuangdao First Hospital,Qinhuangdao 066000,China;Chengde Medical College,Chengde 067000,China)

机构地区：[1]秦皇岛市第一医院肿瘤科,河北秦皇岛066000 [2]承德医学院,河北承德067000

出　　处：《生物技术》2022年第1期62-67,共6页Biotechnology

基　　金：秦皇岛市科技计划项目(201805A138)。

摘　　要：[目的]探讨miRNA-140介导Smad3对非小细胞肺癌(NSCLC)生物学的作用。[方法]Western Blot、RT-PCR法分别检测NSCLC癌组织、正常组织和细胞系A549、95C、NCI-H460及正常支气管上皮细胞系HBE中Smad3、miR-140的表达水平,TargetScan基因软件、双荧光素酶报告实验验证miR-140和Smad3之间的靶向调节关系。取A549、NCI-H460细胞分别转染沉默Smad3蛋白(si-Smad3组)和空白蛋白(si-NC组),比较si-Smad3和si-NC组NSCLC细胞上皮-间充质转化(EMT)、转移率和侵袭能力。[结果]与癌旁和HBE细胞株比较,NSCLC组织及A549、95C、NCI-H460细胞株中Smad3相对表达水平更高(P<0.05),miR-140相对表达水平更低(P<0.05)。miR-140可靶向介导Smad3;A549细胞和NCI-H460细胞中,过表达miR-140组EMT、细胞转移和侵袭能力依次低于miR-NC组低于敲除miR-140组(P<0.05)。[结论]NSCLC组织及细胞株中Smad3表达更高,而miR-140表达水平更低,高水平的miR-140可抑制NSCLC细胞侵袭、转移能力,该信号通路与抑制Smad3蛋白的表达有关。[Objective]To investigate the biological effects of Smad3 mediated by microRNA-140(miR-140)on non-small cell lung cancer(NSCLC)cells.[Method]Western Blot and RT-PCR were used to detect the expression levels of Smad3 and miR-140 in NSCLC tissues,normal tissues,cell lines A549,95 C,NCI-H460 and normal bronchial epithelial cell line HBE,respectively.Targetscan gene software and dual luciferase reporter assay were used to verify the targeted regulatory relationship between miR-140 and Smad3.A549 and NCI-H460 cells were transfected with Smad3 silencing protein(si-Smad3 group)and blank protein(si NC group),respectively.The epithelial mesenchymal transition(EMT),metastasis rate and invasion ability of NSCLC cells in si-Smad3 and si NC groups were compared.[Result]Compared with paracancerous and HBE cell lines,Smad3 expression level was higher in NSCLC tissues and A549,95 C,NCI-H460 cell lines(P<0.05),and miR-140 expression level was lower(P<0.05).miR-140 can target Smad3;In A549 cells and NCI-H460 cells,EMT,cell metastasis and invasion ability of miR-140 overexpression group were lower than those of miR-NC group and miR-140 knockout group(P<0.05).[Conclusion]The expression of Smad3 was higher in NSCLC tissues and cell lines than that of miR-140;high level of miR-140 can inhibit the invasion and metastasis of NSCLC cells,which is related to the inhibition of Smad3 protein expression.

关 键 词：微小RNA-140 SMAD3 非小细胞肺癌 上皮-间充质转化 转移 侵袭 

分 类 号：R734.2[医药卫生—肿瘤]