Identification of hub genes, signaling pathways and immune infiltration of recurrent spontaneous abortion based on bioinformatics analysis with clinical verification

Abstract

ObjectiveThe present study was aimed to identify hub genes associated with recurrent spontaneous abortion (RSA) via both bioinformatics analysis and clinical verification, also to evaluate the related pathways and immune infiltration situation of RSA, for exploring its underlying mechanism. Materials and methodsWe screened candidate hub genes associated with RSA via bioinformatic analysis in the microarray datasets GSE22490 downloaded from the Gene Expression Omnibus (GEO) database. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and Western blotting were used to validate these hub genes. Several kinds of enrichment analysis were carried out to find out the pathways related to RSA. Additionally, CIBERSORT was used for evaluation of local immune Infiltration status of RSA. ResultsThere were 536 differentially expressed genes (DEGs) including 301 upregulated and 235 downregulated genes in RSA group compared with healthy control group. Four hub genes (STAT3, TLR2, TLR4 and CD86) were finally screened out according to the protein–protein interaction (PPI) network analysis, RT-qPCR and Western blotting. Enrichment analysis showed that Toll-like receptor signaling pathway, neutrophil chemotaxis, chemokine signaling pathway and Fc gamma receptor-mediated phagocytosis were strongly associated with RSA. And in immune infiltration analysis, RSA tissue was found containing a higher proportion of monocytes and eosinophils. ConclusionThis study screened out four hub genes and several important pathways changed in the trophoblastic tissue of RSA patients. We also found that monocytes and eosinophils may be involved in RSA. These findings provide theoretical basis for further studies on the mechanisms of RSA.