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、MMP2 mRNA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MMP2 3′-UTR的调控关系。miR-29a mimics(模拟剂)、 miR-29a inhibitor(抑制剂)和miR-NC (对照)转染DU145细胞后分组为、miR-29a mi模拟组、miR-29a in抑制组和miR-NC对照组,检测各组细胞中miR-29a、MMP2 mRNA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、MMP2 mRNA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-29 a on the proliferation, migration and invasion of prostate cancer DU145 cells by regulating the expression of MMP2.Methods The expression of miR-29 a, MMP2 mRNA and protein were detected by qRT-PCR and western blot(WB) in prostate RWPE-1 and DU-145 cells. After transfected with miR-29 a mimics/NC mimics, DU145 cells were divided into the group of miR-29 a mimics, miR-29 a 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-29 a was up-regulated and that of MMP2 mRNA and protein were down-regulated in DU145 cells(P<0.05). After transfected with miR-29 a mimics in DU145 cells, the expression of miR-29 a was increased(P<0.05) and MMP2 mRNA and protein were decreased(P<0.05). After transfected with miR-29 a inhibitor in DU145 cells, the expression of miR-29 a 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-29 a.Overexpression of miR-29 a resulted in significant inhibition of proliferation, migration and invasion of DU145 cells(P<0.05). Conclusions MiR-29 a regulates the expression of MMP2 in DU145 cells, which may affect cell proliferation, migration and invasion.
作者
吴硕
辛士永
张建国
WU Shuo;XIN Shiyong;ZHANG Jianguo(The Clinical Medicine College of Henan University of Science and Technology ( Department of Urology, The First AffiliatedHospital of Henan University of Science and Technology),Luoyung 471003,Henan,China)
出处
《中国性科学》
2019年第4期10-14,共5页
Chinese Journal of Human Sexuality
