miR-376c-3p通过靶向谷氨酸受体相互作用蛋白1影响乳腺癌细胞的增殖和迁移  

miR-376c-3p regulates the proliferation and migration of breast cancer cells via targeting glutamate receptor interacting protein 1

作　　者：王超[1] 高雨婷 刘子瑶 李涛[1] 王晶哲 刘璐 谢岩 孙晓春[1] WANG Chao;GAO Yuting;LIU Ziyao;LI Tao;WANG Jingzhe;LIU Lu;XIE Yan;SUN Xiaochun(School of Medicine,Jiangsu University,Zhenjiang Jiangsu 212013,China)

机构地区：[1]江苏大学医学院,江苏镇江212013

出　　处：《江苏大学学报（医学版）》2021年第5期380-385,共6页Journal of Jiangsu University：Medicine Edition

基　　金：江苏大学科研项目(8421280004)。

摘　　要：目的:探讨miR-376c-3p在乳腺癌中的表达水平、生物学功能及分子调控机制。方法:通过实时荧光定量PCR检测乳腺上皮MCF-10A细胞及乳腺癌细胞中miR-376c-3p表达水平;然后,用miR-376c-3p mimic转染乳腺癌MCF-7与MDA-MB-231细胞,用平板克隆实验、划痕实验以及Transwell实验分别检测其增殖与迁移能力。利用Starbase数据库预测miR-376c-3p的下游靶基因,并通过蛋白质印迹实验进行验证。结果:与人乳腺上皮MCF-10A细胞相比,乳腺癌MCF-7和MDA-MB-231细胞中miR-376c-3p表达水平显著降低(P<0.01);平板克隆实验显示,过表达miR-376c-3p可以显著抑制乳腺癌细胞增殖(P均<0.05);Transwell实验和划痕实验表明,miR-376c-3p表达上调可抑制乳腺癌细胞迁移能力;通过生物信息学预测分析,结果显示miR-376c-3p可以与谷氨酸受体相互作用蛋白1(glutamate receptor interacting protein 1,GRIP1)结合,且蛋白质印迹实验证实过表达miR-376c-3p可以抑制乳腺癌细胞GRIP1表达,同时波形蛋白表达水平明显下调,E-钙黏蛋白表达明显上调。结论:在乳腺癌细胞中,miR-376c-3p表达下调;在体外,过表达miR-376c-3p可显著抑制乳腺癌细胞的增殖和迁移能力,可能通过靶向GRIP1调节乳腺癌细胞上皮间质转化进程。Objective:To investigate the expression level,biological function and molecular regulation mechanism of miR-376c-3p in breast cancer cells.Methods:The expression level of miR-376c-3p in breast epithelial cell MCF-10A and breast cancer cells was detected by qRT-PCR,and then breast cancer cells MCF-7 and MD-MB-231 were transfected with miR-376c-3p mimic.The proliferation was detected by colony formation assay.The migration and invasion were detected by Transwell assay and wound healing assay.Starbase database was used to predict the downstream target gene of miR-376c-3p,and the targeting relationship was verified by Western blotting.Results:Compared with human breast epithelial cell MCF-10A,the expression of miR-376C-3p was significantly down-regulated in breast cancer cells MCF-7 and MDA-MB-231(P<0.01).Colony formation assay showed that overexpression of miR-376c-3p could inhibit the proliferation of breast cancer cells(all P<0.05).Transwell assay and wound healing assay showed that the upregulation of miR-376c-3p inhibited the migration of breast cancer cells.Bioinformatics results showed that miR-376c-3p could bind to glutamate receptor interacting protein 1(GRIP1),and Western blotting assay showed that miR-376c-3p could inhibit the expression of GRIP1 and vimentin,and increase E-cadherin expression in breast cancer cells.Conclusion:The expression of miR-376c-3p was down-regulated in breast cancer cells;overexpression of miR-376c-3p significantly inhibited the proliferation and migration ability of breast cancer cells in vitro,and miR-376c-3p may regulate the epithelial-mesenchymal transition process of breast cancer by targeting GRIP1.

关 键 词：miR-376c-3p 谷氨酸受体相互作用蛋白1 乳腺癌 上皮间质转化 细胞增殖与迁移 

分 类 号：R737.9[医药卫生—肿瘤]