Bioinformatic analysis and validation of ferroptosis in unexplained recurrent spontaneous abortion

Objective:To explore the the role of key genes of ferroptosis in the occurrence and development of unexplained recurrent spontaneous abortion(UR-SA), and to preliminarily identify the potential bio-markers.Methods:The GSE26787 data set was downloaded from Gene Expression Synthesis (GEO)database, and the differentially expressed genes(DEGs)were screened by GEO2R, obtaining ferroptosis-related genes from FerrDb V2 database.The ferroptosisrelated genes in URSA were obtained by intersection of DEGs and ferroptosis genes.The gene ontology(GO)en-richment and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis of ferroptosis-related genes in URSA were performed using DAVID database.The protein interaction network was analyzed using String data-base and Cytascape software to screen the Hub genes.Real-time fluorescence quantitative polymerase chain reac-tion(RT-qPCR)was used to detect the mRNA expression levels of Hub genes in the decidua tissues of patients in the induced abortion group and the URSA group.Results:A total of 55 ferroptosis-related genes in URSA were screened.GO functional enrichment analysis found that ferroptosis-related genes in URSA were mainly enriched in the regulation of macroautophagy, positive regulation of transcription from RNA polymerase Ⅱpromoter, inte-gral components of the membrane and enzyme and p53 receptor binding.The enrichment analysis of KEGG path-way showed that the most obvious enrichment of ferroptosis-related genes in URSA was the FoxO signaling path-way.String database and Cytoscape software were used to screen the key genes of ferroptosis EGFR、SRC、KRAS、MDM2 in URSA.The results of RT-qPCR showed that the expressions of EGFR, SRC and MDM2 in the URSA group were lower than those in the induced abortion group(all P< 0.05).There was no statistically signifi-cant difference in the expression of KRAS between URSA group and induced abortion group (P> 0.05).Conclusion: Ferroptosis-related genes EGFR, SRC and MDM2 can be used as potential biomarkers for diagnosis and treatment of URSA.