miR-375靶向MMP13抑制骨肉瘤细胞的迁移和侵袭

miR-375 Attenuates The Migration and Invasion of Osteosarcoma Cells by Targeting MMP13

目的为了探究miR-375是否通过影响基质金属蛋白酶13(MMP13)的表达来调控骨肉瘤(osteosarcoma,OS)恶性特征。方法用Lipofectamine 3000试剂盒将质粒、miRNA转染至骨肉瘤细胞和HEK293细胞中。实时定量聚合酶链反应(real-time quantitative PCR,RT-qPCR)检测OS患者和OS细胞中miR-375和MMP13的表达。蛋白质印迹法(Western blot)分析OS患者和OS细胞中MMP13蛋白的表达。双荧光素酶法分析miR-375与MMP13的靶向关系。伤口愈合和transwell实验分别分析OS细胞的迁移和侵袭。结果OS组织中miR-375的表达低于正常组织。MMP13在OS组织中表达上调。在OS患者中,MMP13的表达与miR-375呈负相关。与转染miRNA对照的OS细胞相比,转染miR-375模拟物OS细胞的迁移和侵袭明显被抑制。MMP13能部分逆转miR-375对OS细胞迁移和侵袭的抑制作用。结论在OS细胞中,过表达miR-375通过调控MMP13的表达抑制细胞的迁移和侵袭。

Objective To explore whether miR-375 regulates the malignant characteristics of osteosarcoma(OS)by influencing the expression of MMP13.Methods Plasmid DNAs and miRNAs were transfected into OS cells and HEK293 cells using Lipofectamine 3000 reagent.Real-time quantitative polymerase chain reaction was performed to measure the expression of miR-375 and MMP13 in OS patients and OS cells.Western blot was performed to analyze the MMP13 protein in the patients with OS and OS cells.The targeting relationship between miR-375 and MMP13 was analyzed by luciferase assay.Migration and invasion were analysed by heal wound and transwell assays,respectively.Results miR-375 expression in OS tissues was lower than that in normal tissues.The expression of MMP13 was upregulated in OS tissues.MMP13 expression was negatively correlated with miR-375 expression in patients with OS.Migration and invasion were significantly inhibited in OS cells with the miR-375 mimic compared with OS cells with the miRNA control.MMP13 partially reversed the inhibition of migration and invasion induced by miR-375 in the OS cells.Conclusion miR-375 attenuates migration and invasion by downregulating the expression of MMP13 in OS cells.