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微小RNA-129-5p通过抑制Ⅰ型胶原蛋白α1链(COL1A1)来抑制胃癌细胞的侵袭和增殖。

MiR-129-5p suppresses gastric cancer cell invasion and proliferation by inhibiting COL1A1.

作者信息

Wang Quan, Yu Jinhai

机构信息

Department of Gastrointestinal Surgery, the First Hospital of Jilin University, Changchun 130021, Jilin, China.

出版信息

Biochem Cell Biol. 2018 Feb;96(1):19-25. doi: 10.1139/bcb-2016-0254. Epub 2017 May 8.

Abstract

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.

摘要

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。

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