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。

microRNA-34a靶向ZEB1抑制三阴性乳腺癌细胞血管生成拟态

康乐¹, 陶雅军¹, 张媛媛¹, 陈英杰¹, 方明²

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

收稿日期:2019-09-11出版日期:2020-05-30发布日期:2020-05-13
通讯作者:方明E-mail:fangming0411@126.com
作者简介:康乐(1978-),女,讲师,博士.
基金资助:辽宁省自然科学基金（20170540026）


关键词: microRNA-34a, ZEB1, 三阴性乳腺癌, 血管生成拟态
Abstract: Objective To investigate the effect of microRNA-34a(miR-34a) 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-34a in MDA-MB-231 cells. A luciferase reporter assay was utilized to identify the binding site between miR-34a and ZEB1. An in vitro tube formation assay was performed to evaluate the effect of miR-34a overexpression or ZEB1 knockdown on the formation of VM tubes. Western blotting was used to examine the effects of miR-34a overexpression on VM markers and ZEB1 knockdown on the expression of E-cadherin and vimentin. Results miR-34a was significantly upregulated in MDA-MB-231 cells on transfection with miR-34a mimics. miR-34a overexpression could significantly suppress the incidence of VM,as reflected by inhibition in expression of VM markers in MDA-MB-231 cells. miR-34a 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-34a may inhibit the incidence of VM in triple negative breast cancer cells by targeting ZEB1.
Key words: microRNA-34a, ZEB1, triple-negative breast cancer, vasculogenic mimicry
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