微小RNA-9-5p通过靶向作用于亮氨酸拉链肿瘤抑制因子2对三阴性乳腺癌细胞侵袭能力的影响  

MicroRNA-9-5p exerts its effects on the invasive ability of triple-negative breast cancer cells by targeting leucine zipper tumor suppressor 2

作　　者：张洁[1] 冀宏[1] 韩朋[1] 田志胜[2] 李赛男[3] 郭荣[4] Zhang Jie;Ji Hong;Han Peng;Tian Zhisheng;Li Sainan;Guo Rong(Department of Plastic Surgery,the Second Hospital of Hebei Medical University,Shijiazhuang 050000,China;Department of General Surgery,the Second Hospital of Hebei Medical University,Shijiazhuang 050000,China;Department of Breast Surgery,the Fourth Hospital of Hebei Medical University,Shijiazhuang 050011,China;Department of Ultrasonic Diagnosis,Weifang People’s Hospital,Weifang 261000,China)

机构地区：[1]河北医科大学第二医院整形外科,石家庄050000 [2]河北医科大学第二医院普通外科,石家庄050000 [3]河北医科大学第四医院乳腺外科,石家庄050011 [4]山东省潍坊市人民医院超声科,潍坊261000

出　　处：《中华实验外科杂志》2024年第2期247-250,共4页Chinese Journal of Experimental Surgery

基　　金：河北医学科学研究课题(20240030)。

摘　　要：目的观察三阴性乳腺癌(TNBC)细胞中微小RNA(miRNA, miR)-9-5p对亮氨酸拉链肿瘤抑制因子2(LZTS2)的调控作用, 及其对三阴性乳腺癌细胞侵袭能力的影响。方法收集30例2019年4月至2020年4月期间河北医科大学第四医院乳腺外科三阴性乳腺癌及癌旁标本。采用实时荧光定量聚合酶链反应(PCR)方法检测三阴性乳腺癌组织中miR-9-5p及LZTS2基因的表达水平;将MDA-MB-231细胞分为阴性对照组及miR-9-5p模拟物组, 采用双荧光素酶报告基因实验分析LZTS2是否为miR-9-5p靶基因;再将MDA-MB-231细胞分为阴性对照组及miR-9-5p抑制剂组, 采用蛋白印迹实验检测miR-9-5p对LZTS2蛋白表达的影响;采用细胞侵袭实验检测miR-9-5p抑制剂对细胞侵袭能力的影响;随后, 将MDA-MB-231细胞分为miRNA阴性对照+空载体组及miR-9-5p模拟物+LZTS2组, 利用细胞侵袭实验分析miR-9-5p对细胞侵袭能力的影响是否与LZTS2表达有关, 两组间比较采用t检验。结果实时荧光定量PCR结果表明miR-9-5p在三阴性乳腺癌组织中表达(4.45±0.68)明显高于癌旁组织(1.02±0.07, t=8.691, P<0.05);LZTS2在三阴性乳腺癌组织中的表达(0.35±0.11)明显低于癌旁组织(1.01±0.05, t=9.365, P<0.05);相关性分析结果发现miR-9-5p与LZTS2两者表达呈负相关(r=-0.472, P<0.05);报告基因分析结果表明, miR-9-5p模拟物和LZTS2 3’端非编码区(3’UTR)野生型载体共转染组报告基因活性(0.35±0.02)显著低于对照组(1.02±0.04, t=14.980, P<0.05)。蛋白印迹实验结果显示, 转染miR-9-5p抑制剂组LZTS2蛋白的表达水平(1.28±0.05)明显高于阴性对照组(0.23±0.01, t=17.50, P<0.05)。细胞侵袭实验结果表明, miR-9-5p抑制剂组侵袭细胞数[(121.00±9.89)个]明显低于阴性对照组[(300.00±9.89)个, t=12.79, P<0.05];而与LZTS2过表达质粒共转染细胞后, miR-9-5p模拟物组+LZTS2 (306.50±7.77)与miRNA阴性对照+空载体组(310.50±7.78)比较, 差异无统计学意义(t=0.363, P>0.05)�Objective To observe the regulatory effect of microRNA(miRNA,miR)-9-5p on leucine zipper tumor suppressor 2(LZTS2)in triple negative breast cancer(TNBC)cells,as well as its impact on the invasive ability of these TNBC cells.Methods The specimens of triple-negative breast cancer and adjacent tissues were collected from the Department of Breast Surgery,Fourth Hospital of Hebei Medical University,from April 2019 to April 2020.The real-time fluorescent quantitative polymerase chain reaction(PCR)was used to detect the expression levels of miR-9-5p and LZTS2 genes in triple-negative breast cancer tissues.MDA-MB-231 cells were divided into a negative control group and a miR-9-5p mimics group,and whether LZTS2 was a target gene of miR-9-5p was analyzed using a dual luciferase reporter gene experiment.MDA-MB-231 cells were divided into a negative control group and a miR-9-5p inhibitor group,and the effect of miR-9-5p on LZTS2 protein expression was detected using Western blotting.The effect of miR-9-5p inhibitor on cell invasion capability was assessed using a cell invasion assay.Subsequently,MDA-MB-231 cells were divided into miRNA negative control+empty vector group and a miR-9-5p mimics+LZTS2 group,and cell invasion experiments were used to analyze whether the effect of miR-9-5p on cell invasion ability was related to LZTS2 expression.The comparison between the two groups was performed using t-test.Results The real time quantitative PCR showed that the expression of miR-9-5p in TNBC tissues(4.45±0.68)was significantly higher than that in adjacent tissues(1.02±0.07,t=8.691,P<0.05).The expression of LZTS2 in TNBC tissues(0.35±0.11)was significantly lower than that in adjacent tissues(1.01±0.05,t=9.365,P<0.05).The correlation analysis showed that the expression of miR-9-5p and LZTS2 was negatively correlated(r=-0.472,P<0.05).The analysis of reporter gene showed that the co-transfection of miR-9-5p mimic and LZTS23′untranslated regions(3′UTR)wild-type vector group resulted in significantly lower reporter gene activ

关 键 词：微小RNA 三阴性乳腺癌 侵袭 

分 类 号：R737.9[医药卫生—肿瘤]