微小RNA-429过表达对乳腺癌细胞增殖侵袭的影响  

Effects of overexpression of MiR-429 on the proliferation and invasion of breast cancer cells

作　　者：喻建[1] 牟文辉[1] 向英豪 王波涌[2] YU Jian;MOU Wenhui;XIANG Yinghao(Department of General Surgery,Lichuan People’s Hospital,Hubei, Lichuan 445400,China)

机构地区：[1]湖北省利川市人民医院普外科,445400 [2]武汉大学中南医院甲乳外科,武汉市430071

出　　处：《河北医药》2021年第1期20-24,共5页Hebei Medical Journal

摘　　要：目的探讨微小RNA-429(miR-429)过表达对乳腺癌细胞增殖及侵袭的影响,并分析其与Wnt信号通路的可能作用机制。方法选取2017年8月至2018年6月手术治疗的76例乳腺癌患者,切除癌组织及癌旁组织作为乳腺癌组与癌旁组,采用实时定量聚合酶链反应(qRT-PCR)检测2组miR-429表达水平。体外培养乳腺癌细胞株MCF-7,采用脂质体Lipofectamin 2000介导的方法转染细胞,分为空白对照组、阴性转染组以及miR-429 mimics组,倒置荧光显微镜下检测转染效率,qRT-PCR法检测3组细胞中miR-429表达水平;MTT法及平板克隆形成实验检测细胞增殖能力;Transwell法检测细胞侵袭能力;蛋白免疫印迹法(WB)检测ki67、基质金属蛋白酶-2(MMP-2)、基质金属蛋白酶-9(MMP-9)、细胞核β-链蛋白(β-catenin)及Wnt通路相关蛋白表达情况。结果与癌旁组相比,乳腺癌组组织中miR-429表达水平明显降低(P<0.05);与空白对照组和阴性转染组相比,miR-429 mimics组的miR-429表达水平、细胞增殖抑制率显著升高,细胞克隆形成率、细胞侵袭数量及ki67、MMP-2、MMP-9、Wnt、GSK-3β、β-Catenin表达水平均显著降低,差异有统计学意义(P<0.05)。结论miR-429过表达可抑制乳腺癌细胞增殖及侵袭,其作用机制可能与抑制Wnt信号通路有关。Objective To investigate the effects of overexpression of microRNA-429(microRNA-429)on the proliferation and invasion of breast cancer cells,and to analyze its possible action mechanism in Wnt signaling pathway.Methods Ninety-seven patients with breast cancer who underwent surgical treatment in our hospital from August 2017 to June 2018 were enrolled as research subjects.The specimens of resected breast cancer tissues anf para-cancerous tissues were taken as breast cancer group and para-cancerous tissues group,respectively.The expression levels of microRNA-429 were detected by real-time quantitative polymerase chain reaction(qPCR).And MCF-7 cells were cultured in vitro and divided into control group,negative control group and microRNA-429 mimics group.The transfection efficiency was detected by inverted fluorescence microscopy,the expression levels of microRNA-429 were detected by qRT-PCR,and the proliferation ability of breast cancer cells was detected by MTT and plate cloning assay,and the invasive ability of cells was detected by Transwell method.Moreover the expression levels of Ki67,MMP-2,MMP-9,β-catenin and Wnt pathway related proteins were detected by Western Blot(WB).Results As compared with those in para-cancerous tissues group,the expression levels of microRNA-429 in breast cancer tissue group were significantly decreased(P<0.05).As compared with those in blank control group and negative transfection group,the expression levels of microRNA-429 and the inhibition rate of cell proliferation in overexpression of microRNA-429 group were significantly increased(P<0.05).Moreover the cell cloning rate,the number of cell invasion and the expression levels of ki67,MMP-2,MMP-9,Wnt,GSK-3beta and beta-Catenin were significantly decreased(P<0.05).Conclusion The overexpression of miR-429 can inhibit the proliferation,migration,and invasion of breast cancer cells,and its action mechanism may be related to the inhibition of Wnt signaling pathway.

关 键 词：乳腺癌 miR-429 细胞增殖 细胞侵袭 WNT通路 

分 类 号：R737.9[医药卫生—肿瘤]