miR-32-5p靶向ZEB1、ZEB2基因调控乳腺癌增殖和侵袭能力的机制  

MiR-32-5p affects breast cancer proliferation and invasion by interacting with ZEB1,ZEB2

作　　者：俞杨[1] 姚嘉[1] 杨时平[1] 李冠乔[1] YU Yang;YAO Jia;YANG Shi-ping;LI Guan-qiao(Department of Breast Surgery, Hainan Provincial People’s Hospital, Haikou 570311, China)

机构地区：[1]海南省人民医院乳腺外科,海口570311

出　　处：《临床与实验病理学杂志》2021年第3期264-270,共7页Chinese Journal of Clinical and Experimental Pathology

基　　金：海南省自然科学基金(818QN314);海南省自然科学基金青年基金(818QN345)。

摘　　要：目的探讨ZEB1和ZEB2在乳腺癌中的作用及miR-32-5p对乳腺癌调控的分子机制。方法通过Western blot和qRT-PCR检测乳腺癌和癌旁组织中ZEB1、ZEB2和miR-32-5p的表达,筛选出ZEB1和ZEB2高表达的乳腺癌细胞系,采用Western blot和qRT-PCR检测敲减效率。应用克隆形成、迁移和侵袭实验检测敲减ZEB1或ZEB2后对乳腺癌细胞生物学行为的影响,Western blot检测上皮-间质转化相关蛋白(E-cadherin、vimentin)的表达。双荧光素酶报告系统检测miR-32-5p与ZEB1或ZEB23′UTR区域的结合。Western blot、迁移和侵袭实验检测抑制miR-32-5p后对上皮-间质转化相关蛋白、迁移和侵袭的影响。结果乳腺癌组织中ZEB1、ZEB2和miR-32-5p mRNA表达高于癌旁组织。乳腺癌组织中miR-32-5p表达和ZEB1、ZEB2表达呈正相关。MCF-10A、MDA-MB-231和MCF-7三种细胞中,MCF-7细胞ZEB1和ZEB2表达水平较高。敲减ZEB1或ZEB2后,形成的克隆数目减少,迁移和侵袭能力减弱,E-cadherin蛋白表达增加,vimentin蛋白表达降低。ZEB1或ZEB23′UTR区域存在miR-32-5p的结合位点,抑制miR-32-5p会增加迁移和侵袭能力,E-cadherin蛋白表达增加,vimentin蛋白表达降低。增加ZEB1或ZEB2表达会使抑制作用减弱。结论miR-32-5p通过靶向ZEB1和ZEB2调控E-cadherin和vimentin蛋白表达,抑制乳腺癌的增殖、凋亡、迁移和侵袭。Purpose To investigate the role of ZEB1 and ZEB2 in breast cancer,and the molecular mechanism of miR-32-5p on regulation of breast cancer.Methods The expressions of ZEB1,ZEB2 and miR-32-5p in breast cancer and adjacent tissues were detected by Western blot,and qRT-PCR,and breast cancer cell lines with high ZEB1 and ZEB2 expression were screened out,knock-down efficiency was detected by western blot and qRT-PCR.Clone formation,migration and invasion experiments were performed to detect the cell biological behavior after knocking down of ZEB1 or ZEB2.And Western blot was used to detect the expression of epithelial-mesenchymal transition(EMT)-related proteins(E-cadherin,vimentin).The interaction between miR-32-5p and 3′UTR ZEB1 or ZEB2 was detected by the dual luciferase reporter systems.Western blot,migration and invasion experiments were performed to detect the effects of miR-32-5p on EMT-related proteins expression,migration and invasion.Results ZEB1,ZEB2 and miR-32-5p were higher in breast cancer tissues than those in adjacent tissues.The expression of miR-32-5p was positively correlated with ZEB1 and ZEB2 in breast cancer tissues.MCF-7 cells had a higher ZEB1 and ZEB2 expression.When ZEB1 or ZEB2 was knocked down,the number of clones decreased,the migration and invasion capacitydecreased,E-cadherin protein increased,and vimentin protein decreased.3′UTR region of ZEB1 or ZEB2 exists miR-32-5p binding sites.Inhibition of miR-32-5p could increase migration and invasion capacity,increase E-cadherin protein,and decrease vimentin protein.The inhibition was declined when increasing the expression of ZEB1 or ZEB2.Conclusion miR-32-5p regulates the expression of E-cadherin and vimentin by targeting ZEB1 and ZEB2,and inhibits the proliferation,apoptosis,migration and invasion of breast cancer.

关 键 词：乳腺肿瘤 MICRORNA miR-32-5p ZEB1 ZEB2 

分 类 号：R737.9[医药卫生—肿瘤]