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MicroRNAs have been extensively investigated in relation to the epithelial\u2013mesenchymal transition in a range of malignancies. However, there is a paucity of research on extracellular vesicles derived from the eutopic endometrium of adenomyosis and their encapsulated microRNAs. In this study, we investigated the role of microRNA-25-3p derived from extracellular vesicles in inducing macrophage polarization and promoting the epithelial\u2013mesenchymal transition in endometrial epithelial cells of patients with adenomyosis and controls. We obtained eutopic endometrial samples and isolated extracellular vesicles from the culture supernatant of primary endometrial cells. Real-time quantitative PCR analysis demonstrated that microRNA-25-3p was highly expressed in extracellular vesicles, as well as in macrophages stimulated by extracellular vesicles from eutopic endometrium of adenomyosis; and macrophages transfected with microRNA-25-3p exhibited elevated levels of M2 markers, while displaying reduced levels of M1 markers. After co-culture with the above polarized macrophages, endometrial epithelial cells expressed higher levels of N-cadherin and Vimentin, and lower protein levels of E-cadherin and Cytokeratin 7. It was revealed that microRNA-25-3p encapsulated in extracellular vesicles from eutopic endometrial cells could induce macrophage polarization toward M2, and the polarized macrophages promote epithelial\u2013mesenchymal transition in epithelial cells. However, in vitro experiments revealed no significant disparity in the migratory capacity of endometrial epithelial cells between the adenomyosis group and the control group. Furthermore, it was observed that microRNA-25-3p-stimulated polarized macrophages also facilitated the epithelial\u2013mesenchymal transition and migration of endometrial epithelial cells within the control group. Thus, the significance of microRNA-25-3p-induced polarized macrophages in promoting the development of adenomyosis is unclear, and macrophage infiltration alone may be adequate for this process. We emphasize the specificity of the local eutopic endometrial microenvironment and postulate its potential significance in the pathogenesis of adenomyosis.<\/jats:p>","DOI":"10.1093\/molehr\/gaae010","type":"journal-article","created":{"date-parts":[[2024,2,26]],"date-time":"2024-02-26T06:49:31Z","timestamp":1708930171000},"source":"Crossref","is-referenced-by-count":12,"title":["Extracellular vesicle-encapsulated miR-25-3p promotes epithelial\u2013mesenchymal transition and migration of endometrial epithelial cells by inducing macrophage polarization"],"prefix":"10.1093","volume":"30","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-4271-5253","authenticated-orcid":false,"given":"Yue","family":"Hu","sequence":"first","affiliation":[{"name":"Department of Gynecology and Obstetrics, Qilu Hospital (Qingdao), Cheeloo College of Medicine, Shandong University , Qingdao, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6471-9266","authenticated-orcid":false,"given":"Ming","family":"Yuan","sequence":"additional","affiliation":[{"name":"Department of Obstetrics and Gynecology, Shandong Provincial Hospital, Shandong University , Jinan, Shandong, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5819-2007","authenticated-orcid":false,"given":"Lei","family":"Cheng","sequence":"additional","affiliation":[{"name":"Department of Gynecology and Obstetrics, Qilu Hospital (Qingdao), Cheeloo College of Medicine, Shandong University , Qingdao, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1856-1856","authenticated-orcid":false,"given":"Le","family":"Xu","sequence":"additional","affiliation":[{"name":"Department of Obstetrics and Gynecology, Shandong Provincial Hospital, Shandong University , Jinan, 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