子宫腺肌病发病关键基因的生物信息学分析及验证

Bioinformatics Analysis and Verification of the Key Genes in Patho-genesis of Adenomyosis

本研究从GEO(Gene Expression Omnibus)数据库获取子宫腺肌病患者在位子宫内膜和正常子宫内膜芯片数据,通过生物信息学方法筛选差异表达基因(differentially expressed genes,DEGs),并对DEGs进行基因本体(gene ontology,GO)和京都基因与基因组百科全书(Kyoto encyclopedia of genes and genomes,KEGG)富集分析,利用蛋白-蛋白相互作用网络(protein-protein interaction,PPI)筛选关键基因,并在组织标本中对筛选出来的关键基因的表达水平进行验证。本研究一共筛选出509个DEGs(P<0.05,倍数>2)。GO富集分析显示,DEGs主要与内质网蛋白定位、翻译起始、核转录mRNA分解代谢、多组织代谢过程等生物学过程相关。KEGG富集分析显示,DEGs主要富集在核糖体信号通路、氧化磷酸化通路、帕金森病信号通路、阿尔茨海默病信号通路和亨廷顿病信号通路等。PPI相互作用网络筛选出10个关键基因为RPL35、RPL22、RPL39L、RPL26L1、RPS25、RPS29、RPS24、SEC61A1、SEC61G和FAU。实时荧光定量PCR(real-time quantitative PCR,RT-qPCR)和蛋白免疫印迹(Western blotting,WB)结果显示,RPL35、RPS25、FAU等3个基因的mRNA和蛋白相对表达量均在病例组低表达,和对照组比较差异有统计学意义(P<0.05)。本研究认为子宫腺肌病发病可能与在位子宫内膜细胞核糖体蛋白质合成障碍有关,FAU低表达可能与在位子宫内膜细胞抵抗细胞凋亡有关。

In this study,microarray data from both eutopic endometrium of adenomyosis patients and endometrium of controls were obtained from the GEO database.Bioinformatics method was used to screen DEGs,and GO enrichment analysis and KEGG enrichment analysis were conducted for these DEGs.Then,PPI network was applied to screens key genes,and the expression levels of selected key genes were verified in tissue samples.In this study,a total of 509 DEGs were screened(P<0.05,fold change>2).GO enrichment analysis showed that DEGs was mainly related to protein localization to endoplasmic reticulum,translation initiations,nuclear transcribed mRNA catabolic process,and multi-organism metabolic process.KEGG enrichment analysis showed that DEGs were mainly concentrated in ribosome signaling pathway,oxidative phosphorylation pathway,Parkinson disease signaling pathway,Alzheimer disease signaling pathway and Huntington disease signaling pathway.The PPI network selected out 10 key genes as RPL35,RPL22,RPL39 L,RPL26 L1,RPS25,RPS29,RPS24,SEC61 A1,SEC61 G and FAU.Results of RT-qPCR and Western blotting showed that the relative expression levels of mRNA and protein of RPL35,RPS25 and FAU were lower in case group,the differences were statistically significant compared with control group(P<0.05).This study suggestes that the occurrence of adenomyosis is related to the disorder of ribosomal protein synthesis in the eutopic endometrium,and that low expression of FAU may be involved in the resistance to cell apoptosis of eutopic endometrium cells.