Menstrual effluent induces epithelial–mesenchymal transitions in mesothelial cells

• Published in: Human reproduction (Oxford) Vol. 19; no. 1; pp. 21 - 29
• Main Authors: Demir, A.Y., Groothuis, P.G., Nap, A.W., Punyadeera, C., Goeij, A.F.P.M.de, Evers, J.L.H., Dunselman, G.A.J.
• Format: Journal Article
• Language: English
• Published: Oxford Oxford University Press 01.01.2004; Oxford Publishing Limited (England)
• Subjects: Actins - metabolism; Biological and medical sciences; Cadherins - genetics; Cadherins - metabolism; Cell Communication; Cell Movement; Cells, Cultured; Culture Media, Conditioned - pharmacology; DNA-Binding Proteins - genetics; DNA-Binding Proteins - metabolism; Embryology: invertebrates and vertebrates. Teratology; Epithelial Cells - drug effects; Epithelial Cells - metabolism; Epithelial Cells - pathology; E‐cadherin/endometriosis/mesothelial cells/Snail/vimentin; Female; Fundamental and applied biological sciences. Psychology; Genistein - pharmacology; Humans; Indoles - pharmacology; Keratins - metabolism; Menstruation; Omentum - drug effects; Omentum - pathology; Omentum - physiopathology; Phenotype; RNA, Messenger - metabolism; Snail Family Transcription Factors; Sodium Azide - pharmacology; Sulfonamides - pharmacology; Tissue Distribution; Transcription Factors - genetics; Transcription Factors - metabolism; Tubulin - metabolism; Up-Regulation; Vimentin - genetics; Vimentin - metabolism; E‐cadherin/endometriosis/mesothelial cells/Snail/vimentin; Epithelial cell; Vertebrata; E-cadherin/endometriosis/mesothelial cells/Snail/vimentin; Mammalia
• ISSN: 0268-1161; 1460-2350; 1460-2350
• DOI: 10.1093/humrep/deh042

BACKGROUND: Menstrual effluent affects mesothelial cell (MC) morphology. We evaluated whether these changes were consistent with epithelial–mesenchymal transitions (EMT). METHODS: Monolayer cultures of MC were incubated overnight in conditioned media, prepared from cells isolated form menstrual effluent, with or without kinase and ATP inhibitors. Changes in cell morphology were monitored using time‐lapse video microscopy and immunohistochemistry. Effects on the expression of EMT‐associated molecules were evaluated using real‐time RT–PCR and/or Western blot analysis. RESULTS: Incubation in conditioned media disrupted cell–cell contacts, and increased MC motility. The changes were reversible. During the changes the distribution of cytokeratins, fibrillar actin and α‐tubulin changed. Sodium azide, an inhibitor of ATP production, and Genistein, a general tyrosine kinase inhibitor, antagonized these effects. Wortmannin, a phosphatidylinositol 3‐kinase inhibitor, and SU6656, an Src tyrosine kinase inhibitor, only partially antagonized the effect. The expression of Snail and vimentin was markedly up‐regulated, whereas the expression of E‐cadherin was decreased and cytokeratins were altered. CONCLUSIONS: In MC, menstrual effluent initiates a reversible, energy‐dependent transition process from an epithelial to a mesenchymal phenotype. Involvement of the (Src) tyrosine kinase signalling pathway and the changes in the expression of cytokeratins, Snail, vimentin and E‐cadherin demonstrate that the morphological changes are EMT.