摘要
目的:探讨miR-541-3p对乳腺癌细胞的增殖、迁移、侵袭的影响及其作用机制。方法:将miR-con、miR-541-3p、anti-miR-con、anti-miR-541-3p、si-con、si-SPOCD1分别转染至MCF-7细胞中,分别记为miR-con组、miR-541-3p组、anti-miRcon组、anti-miR-541-3p组、si-con组、si-SPOCD1组;将miR-541-3p质粒分别与pcDNA-con、pcDNA-SPOCD1共转染至MCF-7细胞中,分别记为miR-541-3p+pcDNA-con组、miR-541-3p+pcDNA-SPOCD1组;转染均采用脂质体法。RT-qPCR检测miR-541-3p和SPOCD1 mRNA表达水平;Western blot法检测包含1个基因的SPOC结构域(SPOCD1)、细胞周期素D1(CyclinD1)、基质金属蛋白酶2(MMP2)、基质金属蛋白酶9(MMP9)、β-连环蛋白(β-catenin)的表达;MTT检测细胞存活率;Transwell检测细胞迁移和侵袭;双荧光素酶报告基因检测miR-541-3p和SPOCD1的靶向关系。结果:与正常乳腺上皮细胞MCF-10A相比,乳腺癌细胞MCF-7、T47D、Bcap-37中miR-541-3p表达水平显著降低,SPOCD1表达水平显著升高。miR-541-3p过表达和敲减SPOCD1可降低细胞存活率和细胞迁移、侵袭数量,降低CyclinD1、MMP2、MMP9表达水平。miR-541-3p靶向调控SPOCD1并且SPOCD1高表达能部分逆转miR-541-3p高表达对细胞MCF-7增殖、迁移、侵袭的抑制作用。miR-541-3p过表达抑制Wn/tβ-catenin信号通路激活,SPOCD1高表达能部分逆转miR-541-3p高表达对Wn/tβ-catenin信号通路的抑制作用。结论:miR-541-3p过表达可抑制乳腺癌细胞增殖、迁移和侵袭,其机制可能与SPOCD1和Wn/tβ-catenin信号通路相关,并为乳腺癌的治疗提供新思路和新靶点。
Objective:To investigate the effect of miR-541-3p on proliferation,migration and invasion of breast cancer cells and its mechanism.Methods:miR-con,miR-541-3p,anti-miR-con,anti-miR-541-3p,si-con,and si-SPOCD1 were transfected into MCF-7 cells,respectively,and recorded as miR-con group,miR-541-3p group,anti-miR-con group,anti-miR-541-3p group,si-con group,si-SPOCD1 group;miR-541-3p plasmid was co-transfected into MCF-7 cells with pcDNA-con and pcDNA-SPOCD1,respec⁃tively,and recorded as miR-541-3p+pcDNA-con group and miR-541-3p+pcDNA-SPOCD1 group,respectively,transfection was per⁃formed by liposome method.RT-qPCR was used to detect miR-541-3p and SPOCD1 mRNA expression;Western blot assay was used to detect SPOC domain containing 1 gene(SPOCD1),CyclinD1,matrix metalloproteinase 2(MMP2),matrix metalloproteinase 9(MMP9),β-catenin expression;MTT was used to detect cell viability;Transwell was used to detect cell migration and invasion;dual luciferase reporter gene was used to detect the targeting relationship between miR-541-3p and SPOCD1.Results:Compared with nor⁃mal mammary epithelial cells MCF-10A,the expression of miR-541-3p in breast cancer cells MCF-7,T47D and Bcap-37 was signifi⁃cantly decreased,and the expression of SPOCD1 was significantly increased.Overexpression miR-541-3p and knockdown of SPOCD1 reduced cell viability and number of cell migration,invasion,and decreased CyclinD1,MMP2,and MMP9 expression levels.miR-541-3p targeted regulation of SPOCD1,and high expression of SPOCD1 partially reversed the inhibitory effect of high expression of miR-541-3p on proliferation,migration and invasion of MCF-7 cells.Overexpression of miR-541-3p inhibited the activation of Wn/tβ-catenin signaling pathway,and high expression of SPOCD1 partially reversed the effect of high expression of miR-541-3p on the inhibition of Wn/tβ-catenin signaling pathway.Conclusion:Overexpression of miR-541-3p can inhibit the proliferation,migration and inva⁃sion of breast cancer cells.The mechanism may be related to SPOCD1 and Wn
作者
戴睿
林萍
宋精玲
周莹
杨耘
唐莹
DAI Rui;LIN Ping;SONG Jing-Ling;ZHOU Ying;YANG Yun;TANG Ying(Electron Microscope Room,Incubation Center of Scientific and Technological Achievements of Kunming Medical University,Kunming 650500,China)
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2021年第6期694-699,共6页
Chinese Journal of Immunology
基金
云南省科技厅科技计划项目[2018FE001(-021)]。
