The Detection Of Differential Expression Genes And The Screen Of Super-enhancers In Ectopic Endometrial Tissues

Objective:we adopt the RNA-Seq to detect the normal tissue and ectopic tissue with endometriosis.On the one hand,we determine the differentially expressed genes and the pathways affect the pathogenesis of ovarian ectopic endometrial tissues.On the other hand,we screen out the significantly high expressed genes as potential genes that may be regulated by related super-enhancers.Than we adopt ChIP-Seq to detetion the ovarian ectopic endometrial tissues with the H3K27 ac cellmarking and conjoint analysis the RNA-Seq data and ChIP-Seq data,authenticating the special super-enhancers near the significantly high expressed genes.Methods:3 surgical ovarian endometriosis specimens(chocolate cyst)taken from patients of endometriosis in childbearing age between 25-40 years old,taking 3 normal endometrial tissues from women of the childbearing age accepted uterectomy(no endometriosis or adenomyosis were testified by postoperative pathology).Each same weight specimen mixed together in each groups.Using Trizol method to extract total RNA,reverse transcripting of RNA to cDNA and build cDNA libruary.Last we adopt the technology of high-throughput sequencing to test the cDNA and do biological analysis.At the same time,we extract the DNA of endometriosis speciments to make protein and DNA cross-linking.Than we attach the H3K27 ac antibody to immune precipitation the target protein.Last we remove the protein crosslinking and high-throughput sequencing were proceeded after purificating the DNA.We identify the special super-enhancers through ROSE algorithm.Results:The RNA-Seq results suggest significantly gene differential expression between the normal endometrium and ectopic endometrial tissue.GO analysis results suggest the differentially expressed genes mainly involved in biological processes of biological adhesion,cell adhesion,anatomical structure development,cell surface receptor signaling pathway and regulation of cell proliferation.The KEGG pathway analysis indicate that the cell adhesion molecules,cytokine-cytokine receptor interaction and ECM-receptor interaction pathways have significant correlation with EMs.The results of ChIP-Seq indicate 81 special super-enhancers in ectopic endometrial tissue and 272 SEs related genes.We put the SEs genes conjoint analysis with differentially expressed genes from RNA-Seq,we get 43 primary genes.we finally identify the NR5A1,GATA4 and FSHR genes have own SUs near their genetic locus.Conclusion:Endometriosis is an multigenic disease.The pathogenesis of this disease involved in many biological processes and cell pathways.Super-enhancers exist near the Differentially expression pathogenic genes of NR5A1,GATA4 and FSHR and may drive their highly expression.