Screening Of Key Genes For Early Recurrent Pregnancy Loss By Integration Analysis

Objective: DNA methylation is an important epigenetic regulatory factor for gene transcription.Recent studies have confirmed that abnormal DNA methylation may be a potential cause of early recurrent pregnancy loss(ERPL),but the potential mechanism has not been fully elucidated.In this study,the integration analysis of transcriptomics and DNA methylation data for early recurrent pregnancy loss can screen the differentially expressed genes in ERPL and explore possible mechanisms.Methods: Transcriptome gene expression profiling data(GSE22490,GSE76862)and gene methylation profiling data(GSE43256,GSE74738)were downloaded from the Gene Expression Omnibus(GEO).GEO2 R was used to analyze methylated differentially expressed genes(MDEGs).The protein-protein interaction network(PPI)was established by STRING,and the function and enrichment analysis were carried out to the network.Transcription factors(TFs)associated with DNA promoter methylation were predicted to explore possible mechanisms of TF-DNA methylation interactions.The function and enrichment analysis were carried out.44 villus tissues of normal early pregnancy and ERPL patients were collected,and the key genes ERCC2,MLH1,PLEK,and FOS were verified by q RT-PCR.Results: 1.After integrated analysis of gene and DNA methylation profiling data,90 hypermethylation-down regulated genes and 49hypomethylation-up regulated genes were identified.These genes were highly expressed in cells and DNA metabolism,immunity,growth and development and other biological enrichment processes.They were showed to play an important role in DNA damage repair,oxytocin signaling pathway and B cell receptor signaling pathway during the enrichment process.The top key genes of PPI network were ERCC2,MLH1,PLEK,FOS.2.Verified key genes by q RT-PCR.Compared with normal early pregnancy group,the expression of MLH1 and ERCC2 was lower in the ERPL group,while that of PLEK and FOS was higher.The former one was statistically significant(p<0.05).3.TFs-methylated-DEGs regulatory networks predicted that transcription factors,such as SP1,POLR2 A,YY1,DRF2 and PKNOX1 were involved in methylation regulation of DNA promoter with hypermethylation-down regulated genes.Transcription factors SP1,POLR2 A,CREM,CREB1 and ZEB2 were involved in the regulation of genes with hypomethylation-up regulated genes.Among these factors,YY1,FOXP3 and P53 may be related to the expression of MLH1 in the ERPL group.Conclusion: In summary,the study identified possible aberrant MDEGs in ERPL,TFs-MDEGs regulatory networks may be related to what resulted in ERPL,and abnormal expression of MLH1 may be related to ERPL...