The Role And Significance Of14-3-3ζ、ER-beta And SF-1Gene Promoter Region Methylation In The Progression Of Human Adenomyosis

Objective:Multiple studies have implicated that aberrant DNA methylation and proteinexpression in certain genes might be involved in the pathogenesis of humanadenomyosis. Based on our previous work, we speculated that the genes encoding for14-3-3ζ, ER-beta and SF-1might play certain roles in the development of humanadenomyosis. Therefore, the present study plans to investigate the potential roles ofthe promoter methylation and protein expression changes of these genes in thedevelopment of adenomyosis.Methods:Selected the ectopic endometrial tissues of patients with adenomyosis andeutopic endometrial tissues of volunteers without endometriosis, respectively. Theendometrial stromal cells (ESCs) were isolated via enzyme digestion, filtration anddifferential velocity adherent techniques. RT-PCR, WB and methylation-specific PCR(MSP) were used to detect the expression of14-3-3ζ, ER-beta and SF-1and thepromoter methylation situation, respectively. Then CCK-8, Transwell and flowcytometry (FCM) assays were used to detect the potentially functional effects of thesegenes in adenomyosis.Results:Compared with control ESCs:(1) The RT-PCR and WB results showed that theprotein expressions of14-3-3ζ, ER-beta and SF-1were significantly higher in theectopic ESCs;(2) The MSP results showed decreased methylation in the promoterCpG island of14-3-3ζ, ER-beta and SF-1in the ectopic ESCs;(3) Functional assaysshowed that ectopic ESCs showed increased proliferation, invasion and migration, thelower in apoptosis.Conclusion:All of the CpG islands in the gene promoter regions of14-3-3ζ, ER-beta andSF-1presented decreased methylation in the stromal cells of adenomyosis; which wasin accordance with the results that up-regulated protein expression were observed inthese genes. Combined with functional assays between ectopic and control ESCs, we speculated that the aberration of14-3-3ζ, ER-beta and SF-1might be associated withthe pathogenesis of adenomyosis. Nevertheless, further studies would be necessary totest this hypothesis.