血管成形术后再狭窄差异基因分析及机制探索

The analysis of the genetic difference in restenosis after angioplasty aand the exploration of its mechanisms

目的从分子水平探索支架内再狭窄(ISR)的关键基因,以及ISR的发病机制。方法通过基因表达综合(GEO)公共数据库获取GSE46560数据集,利用R语言的limma包分析获得ISR组与对照之间的差异表达基因(DEG),并对外周血的微阵列数据集进行加权基因相关网络分析(WGCNA),以探索ISR相关基因。为了鉴定枢纽基因,对DEG与关键模块中的交集基因进行了功能富集分析、通路分析和蛋白质-蛋白质相互作用(PPI)网络构建。通过蛋白质免疫印迹验证靶基因。结果ISR组和对照组共鉴定出243个DEG,其中有109个上调,有134个下调。WGCNA蓝色模块包含2934个基因,是GSE46560数据集中与ISR相关系数最高的模块。筛选DEG和WGCNA交集基因,有ITPK1、SMG9。GO富集和KEGG分析表明,基因主要富集于蛋白磷酸化、细胞周期调节和细胞增殖,以及细胞衰老和TGF-β信号通路。在Cytoscape中,获得2个枢纽基因,为MCM2、RAD52。结论ITPK1、MCM2、RAD52可能是ISR发病机制特异性相关基因,为ISR的鉴定和治疗提供新的靶点。

Objective From the molecular-level to explore the key genes for in-stent restenosis(ISR)and the pathogenesis of ISR.Methods The GSE46560 dataset was obtained from the Gene Expression Omnibus(GEO)public database,the differentially expressed genes(DEGs)between ISR group and control group were obtained by using limma package analysis in R language,and weighted gene co-expression network analysis(WGCNA)was performed on the microarray dataset of peripheral blood to explore ISR-related genes.To identify hub genes,the functional enrichment analysis,pathway analysis and protein-protein interaction(PPI)network construction were performed on DEGs with intersecting genes in the key modules.Finally,the target genes were verified by Western blot analysis(WB).Results A total of 243 DEGs were identified in the ISR group and control group,of which 109 were up-regulated and 134 were down-regulated.The blue module contained 2934 genes and was the module with the highest correlation coefficient with ISR in the GSE46560 dataset.The screening of DEGs and WGCNA intersecting genes obtained ITPK1 and SMG9.GO enrichment and KEGG analysis showed that genes were mainly enriched in protein phosphorylation,cell cycle regulation and cell proliferation,as well as cellular senescence and TGF-βsignaling pathways.In Cytoscape,2 hub genes were obtained,which were MCM2 and RAD52.Conclusion ITPK1,MCM2 and RAD52 may be the genes that are specifically related to the pathogenesis of ISR,providing new targets for the identification and treatment of ISR.