miR-218-5p靶向TPX2调节p53通路影响肺腺癌恶性进展  

miR-218-5p Targeting TPX2 Regulates p53 Pathway and Inhibits Malignant Progression of Lung Adenocarcinoma

作　　者：许建功[1] Jiangong XU(Department of Thoracic Surgery,The Third Affiliated Hospital of Xinxiang Medical College,Xinxiang 453003,China)

机构地区：[1]新乡医学院第三附属医院胸外科,新乡453003

出　　处：《中国肺癌杂志》2023年第10期721-731,共11页Chinese Journal of Lung Cancer

摘　　要：背景与目的肺腺癌(lung adenocarcinoma,LUAD)是肺癌的一种主要亚型,其治疗与诊断依然是目前的研究热点。靶向Xklp2靶蛋白(targeting protein for Xenopus kinesin-like protein 2,TPX2)在多种癌细胞中高表达,可能与LUAD的发生发展相关。本研究旨在探究TPX2对LUAD细胞恶性进程的影响以及调控机制。方法通过生物信息学分析技术,对癌症基因组图谱(The Cancer Genome Atlas,TCGA)数据库中LUAD组织中基因TPX2的表达情况进行分析。实时荧光定量聚合酶链式反应(quantitative real-time polymerase chain reaction,qRT-PCR)检测人肺正常细胞系和人LUAD细胞系中TPX2和miR-218-5p的表达水平。蛋白质印迹法(Western blot)检测细胞系中TPX2蛋白表达以及其对p53信号通路关键蛋白表达的影响。使用生物信息学预测并通过双荧光素酶报告基因检测验证TPX2与miR-218-5p的关系,细胞活力检测(cell counting kit-8,CCK-8)、细胞克隆形成、细胞划痕、Transwell实验、流式细胞术检测miR-218-5p和TPX2对LUAD细胞功能的影响。结果LUAD细胞中TPX2显著高表达,敲低TPX2可以抑制LUAD细胞的增殖、迁移、侵袭,促进凋亡并产生G_(2)/M期阻滞,并且促进p53信号通路关键蛋白的表达。miR-218-5p是TPX2的上游调控因子,可以抑制其表达。过表达miR-218-5p能消除TPX2高表达导致的恶性发展,抑制LUAD细胞的增殖、迁移等恶性进程以及促进p53信号通路。结论miR-218-5p靶向抑制TPX2表达,并通过p53发挥抑制LUAD细胞恶性发展的作用。Background and objective Lung adenocarcinoma(LUAD)is a major subtype of lung cancer,and its treatment and diagnosis remain a hot research topic.Targeting protein for Xenopus kinesin-like protein 2(TPX2)is highly expressed in a variety of cancer cells and may be associated with the progression of LUAD.This study aimed to investigate the effect of TPX2 on the malignant progression of LUAD cells and the regulatory mechanisms.Methods The expression of gene TPX2 in LUAD tissues from The Cancer Genome Atlas(TCGA)database was analyzed by bioinformatics analysis techniques.Quantitative real-time polymerase chain reaction(qRT-PCR)was used to detect the expression levels of TPX2 and miR-218-5p in human lung normal cell lines and human LUAD cell lines.Western blot was used to detect TPX2 protein expression in cell lines and its effect on the expression of key proteins in the p53 signaling pathway.The relationship between TPX2 and miR-218-5p was predicted using bioinformatics and verified by dual luciferase reporter gene assay.Cell counting kit-8(CCK-8)assay,cell clone formation,cell scratching,Transwell assay,and flow cytometry were used to detect the effects of miR-218-5p and TPX2 on LUAD cell function.Results TPX2 was significantly overexpressed in LUAD cells,and knockdown of TPX2 inhibited LUAD cell proliferation,migration,and invasion,promoted apoptosis and induced G_(2)/M phase block,and promoted the expression of key proteins in the p53 signaling pathway.miR-218-5p,an upstream regulator of TPX2,could inhibit its expression.Overexpression of miR-218-5p eliminated the malignant development caused by high expression of TPX2,inhibited the malignant processes of LUAD cells such as proliferation and migration as well as promoted the p53 signaling pathway.Conclusion miR-218-5p targets and inhibits TPX2 expression and exerts an inhibitory effect on the malignant progression of LUAD cells via p53.

关 键 词：肺肿瘤 TPX2 miR-218-5p P53 转移 凋亡 

分 类 号：R734.2[医药卫生—肿瘤]