miR-877-5p靶向YAP1调控乳腺癌细胞的增殖、迁移侵袭

miR-877-5p targets YAP1 to regulate the proliferation,migration and invasion of breast can⁃cer cells

目的:研究微小RNA(miR)-877-5p对YAP1的调控作用及对乳腺癌细胞的增殖、迁移侵袭的影响。方法:实时荧光定量聚合酶链反应(qPCR)和Western blot检测miR-877-5p、YAP1 mRNA和YAP1蛋白在乳腺癌组织中的表达。利用Lipofectamine^TM 2000试剂将miR-877-5p或si-YAP1转染乳腺癌细胞MCF-7。噻唑蓝(MTT)检测细胞增殖,Transwell小室法检测细胞迁移侵袭,Western blot检测细胞周期蛋白D1(CyclinD1)、p21、基质金属蛋白酶(MMP)-2、MMP-9蛋白表达,双荧光素酶报告实验分析miR-877-5p与YAP1的靶向关系。共转染miR-877-5p和pcDNA-YAP1,观察YAP1过表达对miR-877-5p过表达诱导的MCF-7细胞增殖、迁移侵袭的影响。结果:乳腺癌组织中miR-877-5p表达量明显减少,YAP1 mRNA和YAP1蛋白表达量显著增加(P<0.05)。miR-877-5p过表达或抑制YAP1表达明显降低MCF-7细胞的OD值、迁移细胞数、侵袭细胞数和CyclinD1、MMP-2、MMP-9蛋白水平,并显著提高p21蛋白表达量(P<0.05)。miR-877-5p靶向调控YAP1的表达。YAP1过表达逆转了miR-877-5p过表达对MCF-7细胞增殖、迁移侵袭、CyclinD1、MMP-2和MMP-9蛋白表达的抑制作用,以及对p21蛋白表达的促进作用。结论:miR-877-5p在乳腺癌组织中表达下调,miR-877-5p过表达通过靶向YAP1抑制乳腺癌细胞的增殖、迁移侵袭。

Objective: To study the regulation of microRNA(miR)-877-5p on YAP1 and its influence on the proliferation, migration and invasion of breast cancer cells. Methods: Real-time fluorescent quantitative polymerase chain reaction(qPCR) and Western blot were used to detect the expression of miR-877-5p, YAP1 mRNA and YAP1 protein in breast cancer tissues. Lipofectamine^TM 2,000 reagent was used to transfect miR-877-5p or siYAP1 into breast cancer cell MCF-7. MTT assay was used to detect cell proliferation, Transwell chamber assay was used to detect cell migration and invasion, Western blot was used to detect the expression of cyclin D1, p21,matrix metalloproteinase(MMP)-2 and MMP-9 proteins, dual luciferase report was conducted to analyze the targeting therelationship between miR-877-5p and YAP1. miR-877-5p and pcDNA-YAP1 were co-transfected to observe the effect of YAP1 overexpression on the proliferation, migration and invasion of MCF-7 cells induced by miR-877-5p overexpression. Results: The expression of miR-877-5p in breast cancer tissue was significantly reduced, and YAP1 mRNA and the expression ofYAP1 protein were significantly increased(P< 0.05). Overexpression of miR-877-5p or inhibition of YAP1 expression significantly reduced the OD value of MCF-7 cells, number of migrating MCF-7 cells, number of invaded cells and CyclinD1, MMP-2, MMP-9 protein levels, and significantly increased the expression of p21 protein(P< 0.05). MiR-877-5p targets and regulates the expression of YAP1. YAP1 overexpression reversed the inhibitory effects of miR-877-5p overexpression on MCF-7 cell proliferation, migration and invasion, the expression of CyclinD1, MMP-2 and MMP-9 proteins, as well as the promotion of p21 protein expression. Conclusion: The expression of miR-877-5p is down-regulated in breast cancer tissues. Overexpression of miR-877-5p can inhibit the proliferation, migration and invasion of breast cancer cells by targeting YAP1.