Department of Thoracic Surgery, Guangzhou Panyu District Central Hospital, No. 8, Fuyu East Road, Qiaonan Street, Panyu District, Guangzhou, 511486 People's Republic of China.
Cell Mol Biol Lett. 2019 May 28;24:34. doi: 10.1186/s11658-019-0161-1. eCollection 2019.
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).
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.
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.
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.
微小 RNA(miRNAs)已被报道在癌细胞过程中发挥关键作用,包括增殖、转移和细胞周期进展。我们旨在鉴定可作为非小细胞肺癌(NSCLC)细胞生长和侵袭抑制剂的 miRNAs。
收集并保存在液氮中的 15 对 NSCLC 组织样本和癌旁正常组织。使用 qPCR 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 转染质粒。使用 CCK-8 测定法测定细胞增殖。使用 Western blot 测定法测量内皮间充质转化标志物的蛋白水平。使用 Transwell 测定法评估细胞侵袭能力。TargetScan 用于预测 miR-340 的靶标。进行双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。
miR-340-5p 的表达水平在 NSCLC 组织中比在匹配的癌旁正常组织中更频繁地发现较低。miR-340-5p 的过表达显着抑制 NCI-H1650(一种 NSCLC 细胞系)的增殖和侵袭,而抑制 miR-340-5p 刺激细胞生长。使用 TargetScan,我们预测 ZNF503 可能是 miR-340-5p 的靶标。进一步的机制研究表明,强制表达 ZNF503 可以部分消除 miR-340-5p 介导的 NCI-H1650 细胞活力和侵袭的降低,表明 miR-340-5p 以 ZNF503 依赖的方式抑制细胞生长和侵袭。
我们的研究结果表明,miR-340-5p 通过直接靶向 ZNF503 抑制 NCI-H1650 细胞增殖和侵袭,miR-340-5p 可作为治疗 NSCLC 的潜在治疗靶标。