MicroRNA(miR)-518-3p已显示作为肿瘤抑制因子起作用。本研究旨在探讨miR-518-3p在结直肠癌(CRC)中的作用。miR-518-3p模拟物,模拟阴性对照(NC),miR-518-3p抑制剂,抑制剂-NC,使用脂质体2000将ShRNA-TRIP4和ShRNA-NC载体转染到SW480细胞中。使用CCK-8检测细胞活力。殖民地的形成,细胞侵袭,和细胞迁移通过平板集落形成来评估,Transwell分析,和伤口愈合试验,分别。使用RT-qPCR和Westernblot检测相对mRNA和蛋白水平,分别。鉴定miR-518-3p的靶基因甲状腺激素受体相互作用因子4(TRIP4),并使用双荧光素酶报告基因测定进一步验证。与正常组织相比,CRC患者组织中miR-518-3p水平降低,TRIP4显著升高.用miR-518-3p模拟物或ShRNA-TRIP4转染后,细胞活力以时间依赖性方式降低,和菌落形成率,伤口闭合率,转染细胞的侵袭细胞数量远低于相应的NC和对照组。miR-518-3p过表达或沉默TRIP4显著下调MMP-2和MMP-9的表达。miR-518-3p的敲减有相反的作用,TRIP4被鉴定为miR-518-3p的靶标。miR-518-3p对CRC进展的抑制作用与TRIP4相关。
MicroRNA (miR)-518-3p has been shown to function as a tumor suppressor. This study was conducted to investigate the effects of miR-518-3p in colorectal cancer (CRC). The miR-518-3p mimic, mimic negative control (NC), miR-518-3p inhibitor, inhibitor-NC, ShRNA-TRIP4, and ShRNA-NC vectors were transfected into SW480 cells using Lipofectamine 2000. Cell viability was detected using CCK-8. Colony formation, cell invasiveness, and cell migration were assessed by plate colony formation, Transwell assays, and wound healing assays, respectively. Relative mRNA and protein levels were detected using RT-qPCR and Western blot, respectively. The target gene thyroid hormone receptor interactor 4 (TRIP4) of miR-518-3p was identified and further verified using dual-luciferase reporter assay. Compared with normal tissues, levels of miR-518-3p were decreased and TRIP4 was significantly increased in the tissues from patients with CRC. Following transfection with a miR-518-3p mimic or ShRNA-TRIP4, cell viability decreased in a time-dependent manner, and colony formation rate, wound closure rate, and the number of invasive cells were much lower for the transfected cells than in the corresponding NC and control groups. miR-518-3p overexpression or silencing of TRIP4 significantly down-regulated the expression of MMP-2 and MMP-9. Knockdown of miR-518-3p had the opposite effects, and TRIP4 was identified as a target of miR-518-3p. The inhibitory effects of miR-518-3p on the progressions of CRC are associated with TRIP4.