Qin Jun, Wang Feiran, Jiang Haiyan, Xu Junfei, Jiang Yasu, Wang Zhiwei
Department of General Surgery, Affiliated Hospital of Nantong University Nantong 226001, Jiangsu Province, P. R. China.
Medical College of Nantong University Nantong 226001, Jiangsu Province, P. R. China.
Int J Clin Exp Pathol. 2015 Feb 1;8(2):1328-40. eCollection 2015.
A number of cancers show increased expression of paxillin which plays a central role in tumor progression, including colorectal cancer. However, the mechanisms causing paxillin upregulation remains unclear. In our study, bioinformatics analyses suggested that paxillin is predicted to be a direct target of miR-145. We firstly identified paxillin as a new target of miR-145 and demonstrated that miR-145 inhibits paxillin expression by binding to the paxillin mRNA 3'UTR. Therefore, we assume overexpression of paxillin induced by suppression of miR-145 may promote cell migration and invasion. We detected the expression of paxillin and miR-145 in human colorectal cancer tissues by real-time quantitative PCR. Higher expression of paxillin and lower expression of miR-145 was observed in colorectal cancer tissues than corresponding paracancerous tissue. Moreover, the expression of paxillin was negatively correlated with miR-145 expression. A dual-luciferase reporter assay was used to confirm that paxillin was a direct target of miR-145. In CRC cell lines, overexpression of miR-145 could downregulate paxillin protein expression levels, and ectopic overexpression of miR-145 mimics or inhibitor could inhibit or promote cell migration, invasion, proliferation and clone formation in vitro. Taken together, these data suggested that miR-145 plays a pivotal role in colon cancer through inhibiting cell proliferation migration and invasion, and miR-145 may serve as a tumor suppressor by targeting paxillin gene.
许多癌症中桩蛋白的表达增加,其在肿瘤进展中起核心作用,包括结直肠癌。然而,导致桩蛋白上调的机制仍不清楚。在我们的研究中,生物信息学分析表明桩蛋白被预测为miR-145的直接靶点。我们首先确定桩蛋白是miR-145的新靶点,并证明miR-145通过与桩蛋白mRNA的3'UTR结合来抑制桩蛋白表达。因此,我们推测miR-145受抑制诱导的桩蛋白过表达可能促进细胞迁移和侵袭。我们通过实时定量PCR检测了结直肠癌组织中桩蛋白和miR-145的表达。与相应癌旁组织相比,结直肠癌组织中桩蛋白表达较高,miR-145表达较低。此外,桩蛋白的表达与miR-145的表达呈负相关。采用双荧光素酶报告基因检测法证实桩蛋白是miR-145的直接靶点。在结直肠癌细胞系中,miR-145的过表达可下调桩蛋白的蛋白表达水平,异位过表达miR-145模拟物或抑制剂可在体外抑制或促进细胞迁移、侵袭、增殖和克隆形成。综上所述,这些数据表明miR-145通过抑制细胞增殖、迁移和侵袭在结肠癌中起关键作用,并且miR-145可能通过靶向桩蛋白基因发挥肿瘤抑制作用。