microRNA-34a靶向ZEB1抑制三阴性乳腺癌细胞血管生成拟态  

microRNA-34a inhibits vasculogenic mimicry in triple-negative breast cancer cells by targeting ZEB1

作　　者：康乐[1] 陶雅军[1] 张媛媛[1] 陈英杰[1] 方明[2] KANG Le;TAO Yajun;ZHANG Yuanyuan;CHEN Yingjie;FANG Ming(Department of Physiology and Pathophysiology,College of Basic Medical Science,Medical College of Dalian University,Dalian 116622,China;Department of Nephropathy,The First Affiliated Hospital of Dalian Medical University,Dalian 116011,China)

机构地区：[1]大连大学医学院基础部生理学与病理生理学教研室,辽宁大连116622 [2]大连医科大学附属第一医院肾内科,辽宁大连116011

出　　处：《中国医科大学学报》2020年第5期424-428,共5页Journal of China Medical University

基　　金：辽宁省自然科学基金(20170540026)。

摘　　要：目的探讨microRNA-34a(miR-34a)对三阴性乳腺癌细胞MDA-MB-231血管生成拟态的影响和分子机制。方法以脂质体为载体,将miR-34a模拟物转染入MDA-MB-231细胞,采用实时定量PCR检测miR-34a在MDA-MB-231细胞中的过表达情况;采用双荧光素酶报告基因分析系统检测miR-34a和靶基因ZEB1的靶向结合作用;采用成管实验和Western blotting方法检测过表达miR-34a和敲低ZEB1对MDA-MB-231细胞血管生成拟态形成能力的影响。采用Western blotting方法检测敲低ZEB1对上皮-间质转化标志性分子E-cadherin、Vimentin的影响。结果miR-34a模拟物转染MDA-MB-231细胞后,miR-34a的表达明显上调(P<0.05)。过表达miR-34a显著抑制MDA-MB-231细胞的成管能力和血管生成拟态关键因子的表达。miR-34a靶向调控ZEB1。敲低ZEB1显著抑制MDA-MB-231细胞的成管能力和上皮-间质转化过程。结论miR-34a通过靶向抑制ZEB1抑制三阴性乳腺癌细胞血管生成拟态。Objective To investigate the effect of micro RNA-34 a(mi R-34 a)on the incidence of vasculogenic mimicry(VM)in the MDAMB-231 cells and explore the potential underlying molecular mechanisms.Methods Quantitative real-time PCR was used to confirm the transfection of miR-34 a in MDA-MB-231 cells.A luciferase reporter assay was utilized to identify the binding site between miR-34 a and ZEB1.An in vitro tube formation assay was performed to evaluate the effect of miR-34 a overexpression or ZEB1 knockdown on the formation of VM tubes.Western blotting was used to examine the effects of miR-34 a overexpression on VM markers and ZEB1 knockdown on the expression of E-cadherin and vimentin.Results miR-34 a was significantly upregulated in MDA-MB-231 cells on transfection with miR-34 a mimics.miR-34 a overexpression could significantly suppress the incidence of VM,as reflected by inhibition in expression of VM markers in MDA-MB-231 cells.miR-34 a could directly target the 3’untranslated region of ZEB1.Silencing ZEB1 could,therefore,inhibit VM incidence and epithelial-mesenchymal transition in MDA-MB-231 cells.Conclusion miR-34 a may inhibit the incidence of VM in triple negative breast cancer cells by targeting ZEB1.

关 键 词：microRNA-34a ZEB1 三阴性乳腺癌 血管生成拟态 

分 类 号：R737.9[医药卫生—肿瘤]