子宫肌瘤发病关键基因及信号通路的生物信息学分析与筛选

Identification of key genes and pathways associated with uterine myoma development based on bioinformatics

目的:基于GEO数据库芯片数据应用生物信息学方法进行数据挖掘,探寻子宫肌瘤发病的关键基因及信号通路。方法:从GEO数据库中查询并下载子宫肌瘤(物种为人类)基因表达谱芯片数据,使用R软件进行差异基因的统计学分析,并对筛选出的差异基因进行GO及KEGG富集分析,以及蛋白互作网络分析;应用Cytoscape软件进行信号通路可视化。收集子宫肌瘤及瘤旁组织样本各30例,RT-PCR及Western blot法检测关键基因表达情况,从而对芯片结果进行验证。结果:从GEO数据库中获得了GSE593芯片数据集,筛选出172个差异基因,其中高表达72个,低表达100个。GO富集分析显示,差异基因主要富集于细胞外间隙、细胞外基质、RNA聚合酶Ⅱ启动子转录的负调控、血管生成、促细胞成熟、蛋白质结合等方面;KEGG富集分析显示,差异基因参与的主要信号通路富集于流体剪切应力与动脉粥样硬化信号通路、糖尿病并发症中的AGE-RAGE信号通路、黏着斑激酶信号通路以及EGFR酪氨酸激酶抑制剂耐药性信号通路等。RT-PCR及Western blot结果与芯片结果一致。结论:子宫肌瘤的发病是众多基因及信号通路相互作用的结果,PRKCB在子宫肌瘤的发生发展中可能起着重要作用。

Objective:To investigate the potential genes and pathways associated with uterine myoma through microarray expression profiling data analysis and bioinformatics approaches.Methods:Query and download the gene expression profile microarray data of uterine myoma(species was human)from GEO database,and the biological processes,signaling pathways and network of these genes were statistically analysis using“R”software.30 samples of uterine leiomyoma and 30 samples of adjacent tissues were collected.The expression of key genes was detected by RT-PCR and Western blot,so as to verify the results of the chip.Results:GSE593 chip data were obtained from GEO database.We screened 172 differential genes,of which 72 high expression and 100 low expression.Go enrichment analysis showed that the differential genes were mainly concentrated in extracellular space,extracellular matrix,negative regulation of RNA polymeraseⅡpromoter transcription,angiogenesis,promoting cell maturation,protein binding and so on.KEGG enrichment analysis showed that the main signal pathways involved in differential genes were enriched in fluid shear stress and atherosclerosis signal pathways,AGE-RAGE signal pathways in diabetes complications,focal adhesion kinase signal pathways,and EGFR tyrosine kinase inhibitor resistance signal pathways.The results obtained by RT-PCR and Western blot were consistent with the chip results.Conclusion:The pathogenesis of hysteromyoma is the result of the interaction of many genes and signal pathways.PRKCB may play an important role in the occurrence and development of hysteromyoma.