miR-147a靶向ATF2抑制非小细胞肺癌细胞侵袭和迁移  

miR-147a targeting ATF2 inhibits invasion and migration of non-small cell lung cancer

作　　者：王艾青[1] 郑秀花[1] 刘瑾春 陈玉芳[1] 鲍启德[2] 田振涛 WANG Aiqing;ZHENG Xiuhua;LIU Jinchun;CHEN Yufang;BAO Qide;TIAN Zhentao(Department of Clinical Medicine,Henan Nursing Vocational College,Anyang 455000,China)

机构地区：[1]河南护理职业学院临床医学系,安阳455000 [2]濮阳市安阳地区医院肿瘤科,安阳455000 [3]濮阳市安阳地区医院中医科,安阳455000

出　　处：《中国免疫学杂志》2022年第12期1469-1474,共6页Chinese Journal of Immunology

基　　金：河南省医学科技攻关计划项目(201602369)。

摘　　要：目的:探究miR-147a靶向激活转录因子2(ATF2)对非小细胞肺癌迁移、侵袭和上皮-间质转化的影响。方法:RT-qPCR检测非小细胞肺癌组织和细胞miR-147a和ATF2 mRNA表达,Western blot检测ATF2蛋白表达。采用miR-NC、pc-NC、miR-147a mimic、pc-ATF2质粒分别或联合转染H1703细胞,双荧光素酶报告验证其靶向关系,Transwell检测细胞侵袭,划痕实验检测细胞迁移,Western blot检测上皮钙黏蛋白(E-cadherin)、间质钙黏蛋白(N-cadherin)、波形蛋白(Vimentin)和ATF2蛋白表达。裸鼠分为4组:control组、miR-147a mimic组、pc-ATF2组、miR-147a+pc-ATF2组。裸鼠左腋下皮下注射转染后的非小细胞肺癌H1703细胞,第30天颈椎脱位法处死,完整取出皮下肿瘤,免疫组化检测N-cadherin阳性表达,Western blot检测E-cadherin、N-cadherin和Vimentin蛋白表达。结果:与正常肺组织和细胞相比,非小细胞肺癌组织和细胞miR-147a表达显著下调,ATF2 mRNA和蛋白表达显著上调(P<0.01)。miR-147a靶向下调ATF2表达。与对照组相比,miR-147a mimic组H1703细胞ATF2表达显著下调,侵袭数显著减少,划痕闭合率显著降低,E-cadherin表达显著上调,N-cadherin和Vimentin表达显著下调(P<0.01),共转染pc-ATF2逆转miR-147a mimic对H1703细胞的作用。与对照组相比,miR-147a mimic组移植瘤中N-cadherin阳性细胞百分比降低,E-cadherin表达显著上调,N-cadherin和Vimentin表达显著下调(P<0.01),共转染pc-ATF2逆转miR-147a mimic对H1703移植瘤的作用。结论:miR-147a靶向ATF2可抑制非小细胞肺癌迁移、侵袭和上皮-间质转化。Objective:To investigate effects of miR-147a targeted activating transcription factor 2(ATF2)on migration,invasion and epithelial-mesenchymal transition of non-small cell lung cancer.Methods:miR-147a and ATF2 mRNA expressions of lung cancer tissues and cells were detected by RT-qPCR,and expression of ATF2 protein was detected by Western blot.miR-NC,pc-NC,miR-147a mimic and pc-ATF2 plasmids were transfected into H1703 cells,and targeting relationship was verified by dual luciferase reporter.Cell invasion was detected by Transwell,cell migration was detected by scratch assay,and E-cadherin,N-cadherin,Vimentin and ATF2 protein expressions were detected by Western blot.Nude mice were divided into four groups:control group,miR-147a mimic group,pc-ATF2 group,miR-147a+pc-ATF2 group,and transfected non-small cell lung cancer H1703 cells were subcutaneously injected into left armpit,and sacrificed by cervical dislocation on 30th d.Subcutaneous tumors were completely removed and positive expression of N-cadherin was detected by immunohistochemistry,expressions of E-cadherin,N-cadherin and Vimentin proteins were detected by Western blot.Results:Compared with normal lung tissues and cells,expression of miR-147a was significantly down-regulated,while expressions of ATF2 mRNA and protein were significantly up-regulated in lung cancer tissues and cells(P<0.01).miR-147a targeting down-regulated ATF2 expression.Compared with control group,ATF2 expression was significantly down-regulated,number of invasion was significantly reduced,scratch closure rate was significantly reduced,E-cadherin expression was significantly up-regulated,and N-cadherin and vimentin expressions were significantly down-regulated of H1703 cells in miR-147a mimic group(P<0.01),and co-transfection of pc-ATF2 reversed effect of miR-147a mimic on H1703 cells.Compared with control group,percentage of N-cadherin positive cells was decreased of xenografts in miR-147a mimic group,E-cadherin expression was significantly up-regulated,and N-cadherin and vimentin expres

关 键 词：miR-147a ATF2 非小细胞肺癌 迁移 侵袭 上皮-间质转化 

分 类 号：R734.2[医药卫生—肿瘤]