摘要： 目的 利用生物信息学分析和验证妊娠期糖尿病的关键基因,明确妊娠期糖尿病的发病机制。方法 利用基因表达图谱 (GEO) 数据库在线分析工具GEOR2对数据集GSE49524进行分析,筛选妊娠期糖尿病的差异表达基因,并进行基因本体 (GO) 和京都基因和基因组数据库 (KEGG) 富集分析。利用String和Cytoscape 软件绘制蛋白-蛋白相互作用 (PPI) 网络,筛选妊娠期糖尿病关键基因。利用GSE103552数据集验证关键基因;ELISA法和实时PCR检测妊娠期糖尿病患者 (妊娠糖尿病组) 与正常妊娠患者 (对照组) 外周血COL4A1、P4HA2蛋白和mRNA表达水平。结果 共鉴定出差异表达基因99个,其中表达上调69个,表达下调30个。GO和KEGG富集分析显示,差异表达基因在AGE-RAGE信号通路、TGF-β信号通路;蛋白质消化吸收、胶原三聚体复合物、血小板衍生生长因子、细胞外基质组织结构、内质网等方面显著富集。PPI鉴定出与妊娠期糖尿病相关的5个关键基因为COL4A1、P4HA2、COL1A1、COL3A1和COL5A2。在数据集GSE103552中COL4A1和P4HA2表达差异显著 (均P < 0.05);实时PCR和ELISA法检测结果显示,与对照组比较,妊娠期糖尿病组COL4A1和P4HA2 mRNA及蛋白表达水平增高 (均P < 0.05)。结论 与妊娠期糖尿病相关的5个关键基因为COL4A1、P4HA2、COL1A1、COL3A1和COL5A2。COL4A1和P4HA2表达上调可能促进了妊娠期糖尿病的发生。

基于生物信息学的妊娠期糖尿病关键基因的筛选及验证

王玲玲¹, 谢守军¹, 李爽¹, 郑华川², 任春丽³, 曹秀艳⁴

1. 承德医学院附属医院检验科, 河北 承德 067000;
2. 承德医学院附属医院中心实验室, 河北 承德 067000;
3. 承德医学院附属医院产科, 河北 承德 067000;
4. 承德市双滦区人民医院检验科, 河北 承德 067000

收稿日期:2022-03-11出版日期:2022-10-30发布日期:2022-10-14
通讯作者:郑华川E-mail:285996164@qq.com
作者简介:王玲玲(1989-),女,硕士研究生.
基金资助:河北省医学科学研究重点课题计划(20170874);河北省科技计划(162777102D);承德市科技计划(201804A016)


关键词: 妊娠期糖尿病, 差异表达基因, 生物信息学, 筛选, 验证
Abstract: Objective To analyze and verify the hub genes of gestational diabetes mellitus (GDM) using bioinformatics and to clarify the pathogenesis of GDM.Methods The Gene Expression Omnibus database online analysis tool GEOR2 was used to analyze the GSE49524 dataset and to screen the differentially expression genes in gestational diabetes mellitus.Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed.The protein-protein interaction (PPI) network was mapped using String and Cytoscape software to identify hub genes.The GSE103552 dataset was applied to verify these results.ELISA and real-time PCR were used to detect the protein and mRNA expression levels of COL4A1 and P4HA2 in the peripheral blood of patients with GDM (GDM group) and healthy pregnant patients (control group).Results In total,99 differentially expression genes were identified,of which 69 were up-regulated and 30 were down-regulated.GO and KEGG enrichment analyses showed that differentially expression genes were significantly enriched in the AGE-RAGE,TGF-β,protein digestion and absorption pathways,platelet-derived growth factors,extracellular matrix structure,collagen trimer complex,and endoplasmic reticulum lumen.The PPI network identified five key genes associated with GDM,namely COL4A1,P4HA2,COL1A1,COL3A1,and COL5A2.COL4A1 and P4HA2 were significantly differentially expressed in the validation dataset GSE103552(P < 0.05).The results of real-time PCR and ELISA showed that,compared with the control group,the mRNA and protein expression levels of COL4A1 and P4HA2 were higher in the GDM group (both P < 0.05).Conclusion The five hub genes associated with GDM were COL4A1,P4HA2,COL1A1,COL3A1,and COL5A2.Up-regulation of COL4A1 and P4HA2 may promote the development of GDM.
Key words: gestational diabetes mellitus, differentially expression gene, bioinformatics, screening, validation
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