miR-26a靶向MMP16参与调控胃癌细胞侵袭作用研究

mi R-26a suppresses gastric cancer cell invasion by targeting MMP16

背景与目的:胃癌发生侵袭转移是导致患者预后不良的重要因素。本研究旨在明确mi R-26a在调控胃癌细胞运动侵袭中的作用及其可能机制。方法:通过实时荧光定量聚合酶链反应(real-time fluorescent quantitative polymerase chain reaction,RTFQ-PCR)检测胃癌组织细胞中mi R-26a表达情况,体外通过CCK-8法、平板克隆形成实验和Matrigel-Transwell实验评价mi R-26a对人胃癌细胞增殖、运动和侵袭能力的影响。荧光素酶报告基因系统评估mi R-26a对下游靶基因的调控作用。结果:胃癌组织中mi R-26a表达较癌旁组织明显下降。mi R-26a体外对胃癌细胞增殖无明显影响,但可显著抑制胃癌细胞的运动和侵袭。相反,抑制mi R-26a表达可促进胃癌细胞的侵袭能力。生物信息学分析提示,基质金属蛋白酶16(matrix metalloproteinase 16,MMP16)为mi R-26直接调控靶基因,mi R-26a可抑制胃癌细胞MMP16 mi RNA和蛋白的表达。荧光素酶报告基因检测提示,mi R-26a可与MMP16 m RNA 3’UTR结合诱导其转录后抑制。进一步研究提示,MMP16 si RNA对胃癌细胞侵袭具有模拟mi R-26a作用,而过表达MMP16可拮抗mi R-26a对胃癌细胞侵袭的影响。结论:mi R-26a可通过靶向MMP16来抑制胃癌细胞运动侵袭,mi R-26a可作为抑制胃癌细胞侵袭的重要干预靶点。

Background and purpose： Invasion and metastasis lead to poor prognosis in gastric cancer. In this study, we investigated the potential function of miR-26a in gastric cancer. Methods： Real-time fluorescent quantitative polymerase chain reaction （RTFQ-PCR） was used to detect the expression of miR-26a in gastric cancer cells. In vitro CCK-8 assay, cloning formation assay and Matrigel-Transwell assay were used to evaluate the proliferation, migration and invasion of gastric cancer cells. A luciferase reporter assay was also conducted to confirm that matrix metalloproteinase-16 （MMP16） is a direct target of miR-26a. Results： miR-26a was down-regulated in gastric cancer tissues compared with that in non-cancerous tissues. Functional studies showed that miR-26a inhibited cell proliferation, colony formation, cell motility and invasion. However, miR-26a had no effect on cell proliferation. We also characterized MMP16 as a direct target of miR-26a. We showed that knocking down MMP16 in gastric cancer cells significantly decreased MMP16 expression and inhibited cell invasion, whereas ectopic MMP16 expression significantly abrogated the suppressed cell invasion induced by miR-26a. Conclusion： miR-26a suppresses gastric cancer cell invasion by targeting MMP16. miR-26a could represent a potential therapeutic target for gastric cancer.