The Exploration Of The Relationship Between Estrogen Metabolic Key Genes StAR, NR5A1 And DNA Methylation In Endometriosis

Object:This article explored the relationship between estrogen metabolic key genes St AR, NR5A1 and DNA Methylation in endometriosis, providing new insight to explore the pathogenesis of endometriosis.Method:Firstly, we measured the expressions of DNMTs, TETs, St AR, NR5A1 m RNA in ectopic and eutopic endometrial tissues from endometriosis patients and control endometrial tissues from infertile women by real-time PCR and detected the expressions of DNMT1, DNMT3 A protein by immunohistochemistry. Then through the primary endometriosis stromal cells model, we used demethylating agent 5-aza-2'-deoxycytidine(ADC) as positive control and explored the m RNA expression by real-time PCR and DNA methylation status of St AR, NR5A1 promoter region by bisulfite DNA sequencing after LXA4, E2, IL-1? treated. Finally we detected the effect of LXA4, E2, IL-1? on the m RNA expression of DNMTs, TETs by real-time PCR to explore the probably mechanism of the changes of DNA methylation status.Result:St AR, NR5A1 m RNA were up-regulated significantly and its DNA methylation level were obvious down-regulated in endometriosis, compared with control endometrial and eutopic tissue of patients. There was a negative correlation between St AR, NR5A1 m RNA expression and their DNA methylation level. The Spearman' correlation coefficient of St AR, NR5A1 was-0.7820 and-0.7369 separately. Treating the eutopic endometrial stromal cells with demethylating agent ADC could enhance the m RNA expression of St AR, NR5A1. LXA4, E2, but not IL-1? could promote the m RNA expression of St AR in varying degrees. However the three agents all had not effect on DNA methylation level of their promoter region. Exogenous LXA4, E2 decreased the level of DNA methylation of NR5A1 promotor region while IL-1? had not influence on NR5A1 DNA methylation level. In addition, our research found that DNMT1, 3B and TET1, TET2, TET3 m RNA all decreased in ectopic endometrial tissue, especially the DNMT 1 and 3B. Treated the eutopic endometrial stromal cells with different concentration of LXA4, E2 lead to abnormal expression of some DNMTs and TETs; DNMTs and TETs m RNA expression were all up-regulate after 1 n M LXA4 was treated; DNMT1 TET2 m RNA expression were up-regulated but DNMT3 A, DNMT3 B, TET3 m RNA were down-regulated in eutopic endometrial after 300 n M E2 was treated.Conclusion:There are aberrant DNA methylation status in endometriosis. Administering E2 and LXA4 can destroy the balance between DNMTs and TETs, alter DNA methylation status of St AR, NR5A1 promotor region, augment St AR, NR5A1 m RNA expression, and then enhance the biosynthesis of E2, promoting the development and progression of endometriosis.