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Title: [miR-144-3p Inhibits the Invasion and Metastasis of Lung Adenocarcinoma Cells  by Targeting IRS1].
Author: Bai J, Hu Y, Chen X, Chen L, Zhang L, Yin C, Li H.
Journal: Zhongguo Fei Ai Za Zhi; 2021 May 20; 24(5):323-330. PubMed ID: 34034455.
Abstract:
BACKGROUND: MicroRNAs (miRNAs) are short non-coding RNAs that regulate gene expression, influence cellular processes, and promote disease development. Variations in miRNA expression have been observed in many diseases, including hepatitis, cardiovascular disease, and cancer. The aim of this study is to investigate the effect of miR-144-3p on the invasion and metastasis of lung adenocarcinoma by targeting recombinant insulin receptor substrate 1 (IRS1). METHODS: The expression of miR-144-3p in patients with lung adenocarcinoma was queried through bioinformatics database. MirTarPathway was used to analyze the KEGG enrichment pathway of miRNA. The expression and plasmid transfection efficiency of miR-144-3p in lung adenocarcinoma cell lines were detected by quantitative reverse transcription polymerase chain reaction (qRT-PCR). Transwell assay was used to detect the changes of cell invasion and migration ability in different groups. Bioinformatics determined the key genes (Hub genes) of miR-144-3p; Double luciferase target assay was used to detect the mutual binding of miR-144 and IRS1. Western blot assay was used to detect the expression of IRS1 in different cell lines and the expression of after overexpression of miR-144. RESULTS: The expression of miR-144-3p in lung adenocarcinoma tissues was decreased, qRT-PCR results indicated that the expression of miR-144-3p in lung adenocarcinoma cell A549 was significantly decreased (P<0.05), and the overexpressed plasmid was successfully transfected (P<0.05). Overexpression of miR-144 decreased the ability of cell migration and invasion (P<0.05). The expression of IRS1 was up-regulated in lung adenocarcinoma tissues. Survival analysis showed that patients with lung adenocarcinoma with high IRS1 expression had a poor prognosis (P<0.05). Double luciferase assay results showed that miR-144 could specifically identify 3'-UTR of IRS1 and inhibit reporter enzyme expression (P<0.05). Western blot indicated that the expression of IRS1 was increased in A549 cells (P<0.05). After overexpression of miR-144, the expression level of IRS1 protein was decreased (P<0.05). Transwell experiment proved that miR-144-3p could inhibit invasion and metastasis of lung adenocarcinoma cells by targeting IRS1 (P<0.05). CONCLUSIONS: MiR-144-3p inhibits the invasion and migration of A549 cells through targeted regulation of IRS1, thus playing an anticancer role in tumors. 【中文题目：miR-144-3p通过靶向调控IRS1抑制肺腺癌 细胞的侵袭和转移】【中文摘要：背景与目的 MicroRNAs（miRNAs）是短的非编码RNA，是能够调控基因表达、影响细胞过程、促进疾病发展的重要分子。在许多疾病中可以观察到miRNA表达的变化，包括肝炎、心血管疾病和癌症。本研究旨在探讨miR-144-3p靶向调控胰岛素受体底物1（insulin receptor substrate 1, IRS1）对肺腺癌细胞侵袭和转移的影响。方法通过生物信息学数据库查询miR-144-3p在肺腺癌患者中的表达情况；利用mirTarPathway对miRNA进行KEGG通路富集分析；定量逆转录聚合酶链式反应（quantitative reverse transcription polymerase chain reaction, qRT-PCR）检测miR-144-3p在肺腺癌细胞系中的表达情况与质粒转染效率；Transwell实验检测不同组细胞侵袭迁移能力的变化。生物信息学确定miR-144-3p的关键基因（Hub基因）；双荧光素酶靶标实验检测miR-144和IRS1的相互结合情况；Western blot实验检测IRS1在不同细胞系的表达及过表达miR-144后IRS1的表达情况。结果 miR-144-3p在肺腺癌组织中表达降低，qRT-PCR结果提示miR-144-3p在肺腺癌细胞A549中的表达显著降低（P<0.05）且过表达质粒转染成功（P<0.05）；过表达miR-144后，细胞的迁移、侵袭和增殖能力下降（P<0.05）；IRS1在肺腺癌组织中表达上调；生存分析表明高表达IRS1的肺腺癌患者预后不良（P<0.05）；双荧光素酶实验结果显示，miR-144可特异识别IRS1的3’-UTR抑制报告酶的表达（P<0.05）；Western blot结果提示IRS1在A549细胞中表达升高（P<0.05），过表达miR-144后，IRS1蛋白表达水平下降（P<0.05）；Transwell实验证明，miR-144-3p可通过靶向调控IRS1抑制肺腺癌细胞的侵袭和转移（P<0.05）。结论 miR-144-3p通过靶向调控IRS1抑制A549细胞的侵袭和迁移能力，进而在肿瘤中发挥抑癌作用。】【中文关键词：肺肿瘤；miR-144-3p；侵袭转移；胰岛素受体底物1】.

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