Stanford A型主动脉夹层相关基因KIF20A的共表达网络构建及作用靶点分析

Construction of co-expression network and target analysis of Stanford type A aortic dissection associated gene KIF20A

目的筛选Stanford A型主动脉夹层中的差异表达基因并构建基因共表达网络,以深入解析该疾病的发病机制并筛选潜在的新型分子标志物。方法在GEO数据库获取Stanford A型主动脉夹层相关的基因表达谱芯片,采用R语言limma包筛选差异表达的mRNA(P<0.05、|log2FC|>1),并选取主动脉夹层发病相关的关键差异基因进行深入研究,继而通过实时荧光定量PCR、Western blot检测关键差异基因及其所编码蛋白在Stanford A型主动脉夹层血管组织中的表达情况。利用生物信息学构建该基因的基因共表达网络,并应用R语言ggplot2等包对共表达基因进行功能和通路富集分析,明确该共表达网络所参与的生物学功能。进一步通过对共表达基因进行蛋白-蛋白互作(PPI)网络分析,筛选该关键差异基因的直接相互作用基因,并验证其相关性。最终建立主动脉夹层相关的基因互作图谱,探讨主动脉夹层发病的可能分子机制。两组间比较采用独立样本t检验。结果通过WALD检验筛选出969个在Stanford A型主动脉夹层中差异表达的基因。共表达网络分析发现KIF20A可能作为关键基因在A型主动脉夹层的发病过程中发挥作用,实时荧光定量PCR、Western blot结果显示其在患者组织中表达上调。针对KIF20A的基因共表达网络共纳入53个基因,相关性分析显示其中20个基因表达与KIF20A呈正相关,33个基因表达呈负相关。功能富集分析发现KIF20A共表达网络基因主要富集在上皮、组织等极性的建立和调控、线粒体功能以及精氨酸和脯氨酸代谢等生物学过程。PPI网络分析发现CDK1与KIF20A相互作用,且2个基因的表达量呈正相关(r=0.83,P<0.001)。结论KIF20A基因在Stanford A型主动脉夹层组织中升高,可能通过与CDK1相互作用调控一系列基因表达,影响上皮、组织等的极性建立和调控、线粒体功能等生物学进程,介导主动脉夹层的发生发展。

Objective To further explore the pathogenesis of Stanford Type A aortic dissection and screen potential novel molecular biomarkers by identifying differentially expressed genes in Stanford Type A aortic dissection and constructing a gene co-expression network.Methods Gene expression arrays of Stanford Type A aortic dissection were obtained from the GEO database.The R limma package was used to screen differentially expressed mRNAs(P<0.05,|log2FC|>1).Key differentially expressed genes involved in the pathogenesis of aortic dissection were selected for comprehensive investigation.Subsequently,the expression of key differentially expressed genes and the coded protein were validated by quantitative real-time PCR and Western blot in vascular wall tissues from Stanford Type A aortic dissection patients and control group.The gene co-expression network of the selected gene was constructed by bioinformatic analysis;To clarify the biological functions of the co-expression network,the function and pathway enrichment analysis of the co-expressed genes were performed by ggplot2 package in R language.Further,a protein-protein interaction(PPI)network of these co-expressed genes was constructed to identify the genes interacting with the hub gene directly and validate their correlation.By integrating gene co-expression and PPI networks,we established the gene interaction mechanisms relevant to aortic dissection and explored the specific molecular mechanisms underlying its pathogenesis.Independent sample t test was used for comparison between the two groups.Results A total of 969 differentially expressed genes were identified in Stanford Type A aortic dissection through the application of the WALD test.Based on co-expression network analysis,we selected the upregulated gene KIF20A for further investigation in aortic dissection,and subsequently validated its elevated expression levels in clinical samples.There were 53 genes in the co-expression network of KIF20A,with 20 genes positively correlated and 33 negatively correlated with KIF20A expression.Functional enrichment analysis of this network was mainly enriched in biological processes such as polarity regulation of epithelial cells,mitochondrial function,and arginine and proline metabolism.Further analysis showed that CDK1 directly interacted with KIF20A,and the expression levels of the two genes were positively correlated(r=0.83,P<0.001).Conclusion KIF20A is significantly elevated in Stanford Type A aortic dissection tissues and potentially interacts with CDK1,regulating the expression of a series of genes affecting biological processes such as polarity establishment and regulation of epithelium and tissue,mitochondrial function,and thus mediating the occurrence and development of aortic dissection.