下调miRNA-199a-5p靶向HIF1α激活JAK/STAT3通路促进乳腺癌细胞增殖与恶性转移  

Down-regulated miRNA-199a-5p targeting HIF1αactivates the JAK/STAT3 pathway to promote breast cancer cell proliferation and malignant metastasis

作　　者：徐斌[1] 梁仁杰[1] 刘永萱 卫利民[1] XU Bin;LIANG Ren-jie;LIU Yong-xuan;WEI Li-min(Department of Breast Oncology,the First Affliated Hospital of Henan University of Science and Technology(School of Clinical Medicine,Henan University of Science and Technology,Institute of Oncology,Henan University of Science and Technology),Luoyang 471003,China)

机构地区：[1]河南科技大学第一附属医院乳腺肿瘤外科,河南科技大学临床医学院河南科技大学肿瘤研究所,河南洛阳471003

出　　处：《中国现代普通外科进展》2022年第9期684-689,693,共7页Chinese Journal of Current Advances in General Surgery

基　　金：河南省高等学校重点科研计划项目(20B320005)。

摘　　要：目的:探讨miRNA-199a-5p在乳腺癌MCF-7细胞中的表达及其对细胞增殖与恶性转移的影响。方法:利用RT-qPCR检测miR-199a-5p在乳腺癌细胞(MCF-7细胞)和人正常乳腺上皮细胞(MCF-10A细胞)中的差异表达。过表达或下调miR-199a-5p表达,通过MTT试剂盒检测MCF-7细胞增殖能力,Transwell检测MCF-7细胞的侵袭能力,Western blot检测过表达及下调miR-199a-5p对MCF-7细胞EMT的影响。TargetScan分析miR-199a-5p的作用靶点,双荧光素酶报告基因实验检测miR-199a-5p和HIF1α之间的关系。RT-qPCR检测HIF1α在MCF-7细胞和MCF-10A细胞中的差异表达。HIF1α siRNA质粒转染细胞后,检测下调HIF1α对MCF-7细胞增殖、侵袭和EMT的影响。过表达HIF1α后,Western blot检测细胞内JAK、p-JAK、STAT3和p-STAT3蛋白的表达量。将AG490作用MCF-7细胞后再转染pcDNA-HIF1α质粒,检测MCF-7细胞增殖、侵袭和EMT能力。结果:与MCF-10A细胞相比,MCF-7细胞中miR-199a-5p表达明显降低(P<0.01),HIF1α表达明显上升(P<0.01)。过表达miR-199a-5p抑制了MCF-7细胞增殖、侵袭与EMT,下调miR-199a-5p促进了MCF-7细胞的增殖、侵袭与EMT;miR-199a-5p靶向HIF1α,且负调控HIF1α表达;siRNA下调HIF1α表达后明显抑制了MCF-7细胞增殖、侵袭和EMT;上调HIF1α表达后,激活MCF-7细胞内JAK/STAT3通路,AG490作用MCF-7细胞能明显抑制HIF1α对MCF-7细胞增殖、侵袭和EMT的促进作用。结论:下调miRNA-199a-5p靶向HIF1α激活JAK/STAT3通路促进乳腺癌细胞增殖与恶性转移。Objective:To investigate the expression of miRNA-199a-5p in breast cancer McF-7 cells and its effect on cell proliferation and malignant metastasis.Methods:The differential expression of miR-199a-5p in breast cancer cells(MCF-7 cells)and human normal breast epithelial cells(MCF-10a cells)was detected by RT-qPCR.The proliferation ability of MCF-7 cells was detected by MTT kit,the invasion ability of MCF-7 cells was detected by Transwell,and the effects of overexpression and down-regulation of miR-199a-5p on EMT of MCF-7 cells were analyzed by Western blot.TargetScan was used to analyze the target of miR-199a-5p,and the relationship between miR-199a-5p and HIF1αwas detected by dual-luciferase reporter gene assay.The differential expression of HIF1αin MCF-7 cells and MCF-10A cells was detected by RT-qPCR.The effects of down-regulation of HIF1αon proliferation,invasion and EMT of MCF-7 cells were detected after transfection with HIF1αsiRNA plasmid.After the overexpression of HIF1α,the expression levels of JAK,p-JAK,STAT3 and p-STAT3 were detected by Western blot.MCF-7 cells were treated with AG490 and then transfected with PCDNA-HIF1αplasmid to detect the proliferation,invasion and EMT of McF-7 cells.Results:Compared with MCF-10A cells,the expression of miR-199a-5p in MCF-7 cells was significantly down-regulated(P<0.01),HIF1αexpression was significantly up-regulated(P<0.01).Overexpression of miR-199a-5p inhibited proliferation、invasion and EMT of MCF-7 cells.Down-regulation of miR-199a-5p promoted proliferation,invasion and EMT of MCF-7 cells.miR-199a-5p targeted HIF1αand negatively regulated HIF1αexpression.The proliferation,invasion and EMT of MCF-7 cells were significantly inhibited after the HIF1αexpression was down-regulated.Upregitating HIF1αexpression activated the JAK/STAT3 pathway in MCF-7 cells.AG490 significantly inhibited the proliferation,invasion and EMT of MCF-7 cells induced by HIF1α.Conclusion:Down-regulated miRNA-199a-5p targeting HIF1αactivates the JAK/STAT3 pathway to promote breast

关 键 词：miR-199a-5p HIF1Α JAK/STAT3 乳腺癌 转移 

分 类 号：R737.9[医药卫生—肿瘤]