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miR-330-5p 通过抑制 NOB1 表达抑制非小细胞肺癌细胞生长。

Inhibition of NOB1 by microRNA-330-5p overexpression represses cell growth of non-small cell lung cancer.

机构信息

Department of Thoracic Surgery, The Second Affiliated Hospital of Medical School, Xi'an Jiaotong University, Xi'an, Shaanxi 710004, P.R. China.

Institute of Hepatobiliary Surgery, Southwest Hospital, Third Military Medical University, Chongqing 400038, P.R. China.

出版信息

Oncol Rep. 2017 Oct;38(4):2572-2580. doi: 10.3892/or.2017.5927. Epub 2017 Aug 28.

DOI:10.3892/or.2017.5927
PMID:28849232
Abstract

MicroRNAs (miRNAs) play critical roles in the development and progression of various cancers, including non-small-cell lung cancer (NSCLC). Studies have suggested that miR-330-5p is involved in the progression of several cancers. However, the role of miR-330-5p in NSCLC remains unclear. We investigated the effect on and mechanism of miR-330-5p in the progression of NSCLC. We found that miR-330-5p was significantly downregulated in NSCLC tissues and cell lines as detected by real-time quantitative polymerase chain reaction (RT-qPCR). The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), bromodeoxyuridine (BrdU), colony formation and cell cycle assays showed that overexpression of miR-330-5p markedly inhibited cell growth. Annexin V-FITC/PI and caspase-3 activity assays showed that overexpression of miR-330-5p significantly promoted cell apoptosis of NSCLC cells. Bioinformatics analysis and dual-luciferase reporter assays confirmed NIN/RPN12 binding protein 1 (NOB1) as a target gene of miR-330-5p. RT-qPCR and Western blot analysis showed that overexpression of miR-330-5p inhibited the expression of NOB1 as well as cyclin D1 and cyclin-dependent kinase 4 in NSCLC cells. Moreover, overexpression of NOB1 markedly reversed the miR‑330-5p-mediated inhibitory effect on NSCLC cell growth. Correlation analysis showed that miR‑330-5p expression was inversely correlated with NOB1 mRNA expression in NSCLC tissues. Taken together, our results indicate that miR-330-5p inhibits NSCLC cell growth through downregulation of NOB1 expression. Our study suggests that miR-330-5p may serve as a potential therapeutic target for the treatment of NSCLC.

摘要

微小 RNA(miRNAs)在各种癌症的发展和进展中发挥着关键作用,包括非小细胞肺癌(NSCLC)。研究表明,miR-330-5p 参与了几种癌症的进展。然而,miR-330-5p 在 NSCLC 中的作用尚不清楚。我们研究了 miR-330-5p 在 NSCLC 进展中的作用及其机制。我们通过实时定量聚合酶链反应(RT-qPCR)发现 miR-330-5p 在 NSCLC 组织和细胞系中显著下调。3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐(MTT)、溴脱氧尿苷(BrdU)、集落形成和细胞周期检测表明,miR-330-5p 的过表达显著抑制了细胞生长。Annexin V-FITC/PI 和 caspase-3 活性检测表明,miR-330-5p 的过表达显著促进了 NSCLC 细胞的凋亡。生物信息学分析和双荧光素酶报告基因检测证实 NIN/RPN12 结合蛋白 1(NOB1)是 miR-330-5p 的靶基因。RT-qPCR 和 Western blot 分析表明,miR-330-5p 的过表达抑制了 NSCLC 细胞中 NOB1 以及细胞周期蛋白 D1 和细胞周期蛋白依赖性激酶 4 的表达。此外,NOB1 的过表达显著逆转了 miR-330-5p 对 NSCLC 细胞生长的抑制作用。相关性分析表明,miR-330-5p 的表达与 NSCLC 组织中 NOB1 mRNA 的表达呈负相关。综上所述,我们的研究结果表明,miR-330-5p 通过下调 NOB1 的表达抑制 NSCLC 细胞的生长。我们的研究表明,miR-330-5p 可能作为治疗 NSCLC 的潜在治疗靶点。

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