miR-301a对三阴性乳腺癌细胞侵袭与增殖的调控作用  

Regulation of miR-301a on invasion and proliferation of triple negative breast cancer cells

作　　者：刘恒[1] 王钢乐[1] 李秀楠[1] LIU Heng;WANG Gangyue;LI Xiunan(Department of Breast Surgery, Beijing Obstetrics and Gynecology Hospital, Capital Medical University, Beijing 100006, China)

机构地区：[1]首都医科大学附属北京妇产医院乳腺科,北京100006

出　　处：《西部医学》2021年第5期650-654,共5页Medical Journal of West China

基　　金：首都医科大学附属北京妇产医院中青年学科骨干培养专项(FCYY201816)。

摘　　要：目的探讨microRNA-301a(miR-301a)对三阴性乳腺癌(TNBC)细胞的侵袭、增殖的调控作用及潜在的作用机制。方法应用real-time PCR检测miR-301a在MDA-MB-231细胞与MCF-10A细胞中的表达;转染MDA-MB-231,按转染质粒分为Mimics miR-301a组、Mimics NC组、Inhibitor miR-301a组、Inhibitor NC组,通过Transwell小室实验、MTT实验检测miR-301a对MDA-MB-231细胞侵袭及增殖能力的影响;应用生物信息学软件预测miR-301a的下游靶点,并应用荧光素酶报告实验以及Western blot分析miR-301a对靶点的调节作用。结果miR-301a在MDA-MB-231细胞中的表达明显高于正常乳腺细胞(P<0.05);Transwell小室实验提示过表达miR-301a能够增强MDA-MB-231细胞的侵袭能力(P<0..05);MTT实验提示过表达miR-301a能够增强MDA-MB-231细胞的增殖能力(P<0.05);荧光素酶报告实验以及western blot实验证实miR-301a与MEOX2的靶向关系,Mimics miR-301a组MEOX2蛋白表达量明显低于Mimics NC组和空白组(P<0.05),Inhibitor miR-301a组MEOX2蛋白表达量明显高于Inhibitor NC组和空白组(P<0.05)。结论miR-301a在TNBC中高表达,并可通过靶向作用于MEOX2促进TNBC细胞的侵袭、增殖,从而作为TNBC的促癌基因发挥作用。Objective To investigate the regulatory effects of miR-301a on invasion and proliferation and potential mechanism in the triple negative breast cancer.Methods The expression of miR-301a in cell lines MDA-MB-231 and MCF-10A were determined by real-time PCR.Transfected MDA-MB-231 and set up NC controls,Transwell assay and WTT assay were used to detect the effect of miR-301a on cell invasion and proliferation.The downstream targets of miR-301a was predicted by bioinformatical tools.The regulation between miR-301a and targets was verified by Luciferase reporter assay and Western blot.Results The expression of miR-301a in MDA-MB-231 was significantly higher than that in MCF-10A(P<0.05).The Transwell assay and MTT assay revealed that overexpression of miR-301a could enhance the invasion and proliferation ability of MDA-MB-231.Luciferase reporter assay and western blot indicated that miR-301a inhibited the luciferase activity of MEOX2.Overexpression of miR-301a reduces MEOX2 protein levels(P<0.05),and inhibition of miR-301a expression can increase MEOX2 protein levels(P<0.05).Conclusion miR-301a is high-expressed in TNBC cells and is a cancer promoter which can enhance the invasion and proliferation of TNBC cells through targeting MEOX2.

关 键 词：三阴性乳腺癌 microRNA-301a MEOX2 细胞侵袭与增殖 调控作用 

分 类 号：R737.9[医药卫生—肿瘤]