MiR-29a通过调节MMP2参与前列腺癌DU145细胞的增殖和侵袭能力的研究

摘要

目的 研究miR-29a是否调节MMP2的表达参与前列腺癌DU145细胞的迁移、侵袭和增殖过程.方法通过实时荧光定量(qRT-PCR)和免疫印迹法(WB)检测在正常前列腺上皮RWPE-1细胞和激素非依赖性前列腺癌DU-145细胞内miR-29a、MMP2mRNA和蛋白的表达水平.采用双荧光素酶报告基因实验验证miR-29a与MMP23'-UTR的调控关系.miR-29amimics(模拟剂)、miR-29ainhibitor(抑制剂)和miR-NC(对照)转染DU145细胞后分组为、miR-29ami模拟组、miR-29ain抑制组和miR-NC对照组,检测各组细胞中miR-29a、MMP2mRNA和蛋白的表达;细胞划痕实验和侵袭实验(Transwell小室)检测各组细胞迁移和侵袭;增殖实验(CCK-8)实验检测各组细胞增殖.结果 与RWPE-1细胞相比,DU145细胞中miR-29a的表达减少、MMP2mRNA和蛋白的表达增加.miR-29a模拟剂转染细胞后,miR-29a表达升高,MMP2表达降低;miR-29a抑制剂转染细胞后,miR-29a表达降低,MMP2表达升高.双荧光素酶实验验证了MMP2为miR-29a的靶基因.MiR-29a过表达能抑制DU145细胞的迁移、侵袭和增殖;MiR-29a抑制表达能促进DU145细胞的迁移、侵袭和增殖.结论 在前列腺癌DU145细胞中,miR-29a调节MMP2的表达,进而参与细胞的迁移、侵袭和增殖过程.%Objective To investigate the effect of miR- 29a on the proliferation,migration and invasion of prostate cancer DU145 cells by regulating the expression of MMP2. Methods The expression of miR- 29a,MMP2 mRNA and protein were detected by qRT-PCR and western blot (WB) in prostate RWPE- 1 and DU- 145cells. After transfected with miR- 29a mimics /NC mimics,DU145 cells were divided into the group of miR- 29a mimics,miR- 29a inhibitor,miR-NC. The expression was also analyzed by qRT-PCR and WB. Cell counting kit- 8 (CCK- 8) was used to evaluate cell proliferation and Transwell and Scratch healing assay was performed to assess cell migration and invasion. Results Compared with RWPE- 1 cells,the expression ofmiR- 29a was up-regulated and that of MMP2 mRNA and protein were down-regulated in DU145 cells(P ＜ 0. 05). After transfected with miR- 29a mimics in DU145 cells,the expression of miR- 29a was increased(P ＜ 0. 05) and MMP2 mRNA and protein were decreased(P＜ 0. 05). After transfected with miR- 29a inhibitor in DU145 cells,the expression of miR- 29a was decreased(P ＜ 0. 05) and MMP2 mRNA and protein were increased(P ＜ 0. 05). Dual luciferase assay confirmed that MMP2 was the target gene of miR- 29a. Overexpression of miR- 29a resulted in significant inhibition of proliferation,migration and invasion of DU145 cells (P ＜ 0. 05). Conclusions MiR- 29a regulates the expression of MMP2 in DU145 cells,which may affect cell proliferation,migration and invasion.