背景:据报道,微小RNA(miRNA)在癌细胞过程(包括增殖,转移和细胞周期进程)中起着至关重要的作用。我们旨在鉴定可以在非小细胞肺癌(NSCLC)中抑制细胞生长和侵袭的miRNA。 方法:收集15对成对的NSCLC组织样品和癌周周围正常组织并保存在液氮中。使用qPCR分析检测miR-340-5p和ZNF503 mRNA的表达水平。根据制造商的方案,使用Lipofectamine 3000进行质粒的转染。使用CCK-8测定法测定细胞增殖。内皮-间质转化标志物的蛋白质水平使用蛋白质印迹测定法进行测量。使用transwell测定法评估细胞侵袭能力。 TargetScan用于预测miR-340的靶标。进行了双重荧光素酶报告基因测定,以确认miR-340-5p和ZNF503之间潜在的直接相互作用。 结果:经常发现miR-340-5p在非小细胞肺癌组织中的表达水平低于匹配的癌旁正常组织。 miR-340-5p的过表达显着抑制NCI-H1650(NSCLC细胞系)的增殖和侵袭,而抑制miR-340-5p则刺激细胞生长。使用TargetScan,我们预测ZNF503可能是miR-340-5p的靶标。进一步的机理研究表明,ZNF503的强制表达可以部分消除miR-340-5p介导的NCI-H1650细胞活力和侵袭性的降低,这表明miR-340-5p以ZNF503依赖性的方式抑制了细胞的生长和侵袭。 结论:我们的发现表明,miR-340-5p通过直接靶向ZNF503抑制NCI-H1650细胞的增殖和侵袭,而miR-340-5p可以作为治疗NSCLC的潜在靶点。**
MicroRNA-340-5p suppresses non-small cell lung cancer cell growth and metastasis by targeting ZNF503
Background: MicroRNAs (miRNAs) have been reported to play crucial roles in cancer cell processes, including proliferation, metastasis and cell cycle progression. We aimed to identify miRNAs that could act as suppressors of cell growth and invasion in non-small cell lung cancer (NSCLC). Methods: Fifteen paired NSCLC tissue samples and pericarcinomatous normal tissues were collected and preserved in liquid nitrogen. The expression levels of miR-340-5p and ZNF503 mRNA were detected using a qPCR assay. The transfection of plasmids was conducted using Lipofectamine 3000 according to the manufacturer's protocol. Cell proliferation was determined using a CCK-8 assay. The protein levels of endothelial-mesenchymal transition markers were measured using a western blot assay. Cell invasive ability was evaluated using a transwell assay. TargetScan was used to predict targets of miR-340. A dual luciferase reporter assay was performed to confirm a potential direct interaction between miR-340-5p and ZNF503. Results: The expression level of miR-340-5p was frequently found to be lower in NSCLC tissues than in matched pericarcinomatous normal tissues. Overexpression of miR-340-5p significantly inhibited the proliferation and invasion NCI-H1650 (a NSCLC cell line), while inhibition of miR-340-5p stimulated cell growth. Using TargetScan, we predicted that ZNF503 could be a target of miR-340-5p. Further mechanistic studies demonstrated that the forced expression of ZNF503 could partially abrogate the miR-340-5p-mediated decrease in NCI-H1650 cell viability and invasion, suggesting that miR-340-5p suppressed cell growth and invasion in a ZNF503-dependent manner. Conclusion: Our findings indicate that miR-340-5p inhibits NCI-H1650 cell proliferation and invasion by directly targeting ZNF503 and that miR-340-5p can serve as a potential therapeutic target for treating NSCLC.
pmid: 31160893 Cell Mol Biol Lett 影响因子: 3.367 发表日期: 20190101 官网 免费下载