Wang Quan, Yu Jinhai
Department of Gastrointestinal Surgery, the First Hospital of Jilin University, Changchun 130021, Jilin, China.
Biochem Cell Biol. 2018 Feb;96(1):19-25. doi: 10.1139/bcb-2016-0254. Epub 2017 May 8.
Gastric cancer (GC) is one of the most lethal cancers worldwide. In this study, we aimed to explore the role of miR-129-5p, a newly identified miR-129 member, in GC cells as well as the potential mechanism of action. The results of reverse transcription - qualitative polymerase chain reaction (RT-qPCR) and Western Blot showed that miR-129 was downregulated in GC cells compared with normal ones. Using MTT, colony formation, wound healing assay, and a Transwell assay, we evaluated the proliferation, migration, and invasion abilities of transfected cells, and confirmed miR-129-5p as a tumor suppressor in GC. After a microarray analysis comparing different gene expressions in miR-129-5p transfected SGC-7901 cells, COL1A1 was selected for biggest fold-change and potential target of miR-129-5p predicted by TargetScan. Measured by RT-qPCR and Western blot, COL1A1 turned out to be upregulated in GC tissues and cells. We further confirmed the targeting relationship between miR-129-5p and COL1A1 by dual luciferase assay. By manipulating the expression of COL1A1 in SGC-7901 cells, cell proliferation, migration, and invasion were examined and the tumor-promoting function of COL1A1 was validated. Moreover, co-transfection of miR-129-5p mimics and COL1A1 attenuated the tumor-promoting effects induced by a single-transfection of COL1A1, and miR-129-5p inhibitor counteracted the tumor-suppressing effects of COL1A1 siRNA. Collectively, the data demonstrate the important functions of the miR-129-5p-COL1A1 axis in GC: miR-129-5p suppresses GC cell proliferation, migration, and invasion, by selectively inhibiting COL1A1. This study provides new therapeutic targets for the clinical treatment of GC.
胃癌(GC)是全球最致命的癌症之一。在本研究中,我们旨在探讨新鉴定的miR - 129成员miR - 129 - 5p在胃癌细胞中的作用及其潜在作用机制。逆转录 - 定量聚合酶链反应(RT - qPCR)和蛋白质免疫印迹结果显示,与正常细胞相比,胃癌细胞中miR - 129表达下调。通过MTT法、集落形成实验、伤口愈合实验和Transwell实验,我们评估了转染细胞的增殖、迁移和侵袭能力,并证实miR - 129 - 5p在胃癌中起到肿瘤抑制作用。在对miR - 129 - 5p转染的SGC - 7901细胞中不同基因表达进行微阵列分析后,根据TargetScan预测,COL1A1因最大倍数变化及潜在的miR - 129 - 5p靶标而被选中。通过RT - qPCR和蛋白质免疫印迹检测发现,COL1A1在胃癌组织和细胞中上调。我们通过双荧光素酶报告基因实验进一步证实了miR - 129 - 5p与COL1A1之间的靶向关系。通过调控SGC - 7901细胞中COL1A1的表达,检测细胞增殖、迁移和侵袭情况,并验证了COL1A1的促肿瘤功能。此外,共转染miR - 129 - 5p模拟物和COL1A1可减弱COL1A1单转染诱导的促肿瘤作用,而miR - 129 - 5p抑制剂可抵消COL1A1 siRNA的肿瘤抑制作用。总体而言,数据表明miR - 129 - 5p - COL1A1轴在胃癌中具有重要作用:miR - 129 - 5p通过选择性抑制COL1A1来抑制胃癌细胞的增殖、迁移和侵袭。本研究为胃癌的临床治疗提供了新的治疗靶点。