肺腺癌ceRNA调控网络构建及AFAP1-AS1功能验证  

Foundation of ceRNA networks and functional validation of AFAP1-AS1 in lung adenocarcinoma

作　　者：王会新 李倩 侯晓雯 史新竹 冯旭[1] WANG Huixin;LI Qian;HOU Xiaowen;SHI Xinzhu;FENG Xu(Statistics Teaching and Research Office of the School of Public Health,Shenyang Medical College,Shenyang,110034,P.R.China;Epidemiology Teaching and Research Office of the School of Public Health,Shenyang Medical College,Shenyang,110034,P.R.China)

机构地区：[1]沈阳医学院公共卫生学院统计教研室,沈阳110034 [2]沈阳医学院公共卫生学院流行病学教研室,沈阳110034

出　　处：《中国胸心血管外科临床杂志》2024年第4期576-584,共9页Chinese Journal of Clinical Thoracic and Cardiovascular Surgery

基　　金：辽宁省科学技术计划项目(2019-ZD-0324);沈阳医学院研究生科技创新基金项目(Y20210509)。

摘　　要：目的基于生物信息学方法构建肺腺癌(lung adenocarcinoma,LUAD)特异性长链非编码RNA(long non-coding RNA,lncRNA)相关的竞争性内源RNA(competing endogenous RNA,ceRNA)调控网络,并分析肌动蛋白纤维相关蛋白1-反义RNA1(actinfilament-associated protein 1-antisense RNA1,AFAP1-AS1)在LUAD中的功能机制,以期为LUAD治疗靶点研究提供新的方向。方法从基因表达综合性基因库(GEO)中下载LUAD的基因芯片,并使用GEO2R在线软件筛选LUAD与正常组织之间具有差异表达的lncRNA和mRNA,并通过在线数据库预测其靶基因,构建ceRNA网络,并进行富集分析。在细胞实验中,对AFAP1-AS1进行基因敲除并构建siRNA,通过细胞转染法转染LUAD细胞A549。用CCK8、Transwell、划痕实验和流式细胞术实验检测细胞增殖、侵袭、迁移和凋亡的能力。结果在LUAD的芯片中共鉴定出6个差异表达lncRNA和494个差异表达的mRNA。ceRNA网络共涉及6个lncRNA、22个miRNA和55个mRNA。富集分析发现mRNA与癌症相关通路有关。在细胞实验中,敲除AFAP1-AS1可抑制细胞增殖、侵袭和迁移,且AFAP1-AS1促进细胞凋亡。结论本研究构建了一个lncRNA介导的ceRNA网络,这可能有助于进一步研究LUAD的作用机制。此外,通过细胞实验,本研究发现AFAP1-AS1有潜力作为LUAD的治疗靶点。Objective A competing endogenous RNA(ceRNA)regulatory network associated with long noncoding RNA(lncRNA)specific for lung adenocarcinoma(LUAD)was constructed based on bioinformatics methods,and the functional mechanism of actinfilament-associated protein 1-antisense RNA1(AFAP1-AS1)in LUAD was analyzed,in order to provide a new direction for the study of LUAD therapeutic targets.Methods The gene chip of LUAD was downloaded from the Gene Expression Omnibus(GEO),and lncRNA and mRNA with differential expression between LUAD and normal tissues were screened using GEO2R online software,and their target genes were predicted by online databases to construct ceRNA networks and perform enrichment analysis.In cell experiments,AFAP1-AS1 was genetically knocked down and siRNA was constructed and transfected into LUAD cells A549 by cell transfection.CCK8,transwell,scratch assay and flow cytometry were used to detect the ability of cells to proliferate,invade,migrate and apoptosis.Results A total of 6 differentially expressed lncRNA and 494 differentially expressed mRNA were identified in the microarray of LUAD.The ceRNA network involved a total of 6 lncRNA,22 miRNA,and 55 mRNA.Enrichment analysis revealed that mRNA was associated with cancer-related pathways.In cell assays,knockdown of AFAP1-AS1 inhibited cell proliferation,invasion,and migration,and AFAP1-AS1 promoted apoptosis.Conclusion In this study,we construct a lncRNA-mediated ceRNA network,which may help to further investigate the mechanism of action of LUAD.In addition,through cellular experiments,AFAP1-AS1 is found to have potential as a therapeutic target for LUAD.

关 键 词：生物信息学 肺腺癌 竞争性内源RNA 长链非编码RNA 肌动蛋白纤维相关蛋白1-反义RNA1 

分 类 号：R734.2[医药卫生—肿瘤]