title: Menstrual effluent induces epithelial-mesenchymal transitions in mesothelial cells.
abstract: 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 alpha-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.
authors.affiliation: Department of Obstetrics & Gynaecology, Research Institute Growth and Development (GROW), Academisch ziekenhuis Maastricht and Maastricht University, Maastricht, The Netherlands.
authors.forename: A Y
authors.initials: AY
authors.lastname: Demir
authors.forename: P G
authors.initials: PG
authors.lastname: Groothuis
authors.forename: A W
authors.initials: AW
authors.lastname: Nap
authors.forename: C
authors.initials: C
authors.lastname: Punyadeera
authors.forename: A F P M
authors.initials: AF
authors.lastname: de Goeij
authors.forename: J L H
authors.initials: JL
authors.lastname: Evers
authors.forename: G A J
authors.initials: GA
authors.lastname: Dunselman
mesh_terms: D000199:Actins / Q000378:metabolism; D015820:Cadherins / Q000235:genetics / Q000378:metabolism; D002450:Cell Communication; D002465:Cell Movement; D002478:Cells, Cultured; D017077:Culture Media, Conditioned / Q000494:pharmacology; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D004847:Epithelial Cells / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D005260:Female; D019833:Genistein / Q000494:pharmacology; D006801:Humans; D007211:Indoles / Q000494:pharmacology; D007633:Keratins / Q000378:metabolism; D008598:Menstruation; D009852:Omentum / Q000187:drug effects / Q000473:pathology* / Q000503:physiopathology; D010641:Phenotype; D012333:RNA, Messenger / Q000378:metabolism; D000071250:Snail Family Transcription Factors; D019810:Sodium Azide / Q000494:pharmacology; D013449:Sulfonamides / Q000494:pharmacology; D014018:Tissue Distribution; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D014404:Tubulin / Q000378:metabolism; D015854:Up-Regulation; D014746:Vimentin / Q000235:genetics / Q000378:metabolism
chemical_list: D000199:Actins; D015820:Cadherins; D017077:Culture Media, Conditioned; D004268:DNA-Binding Proteins; D007211:Indoles; D012333:RNA, Messenger; C416927:SU 6656; D000071250:Snail Family Transcription Factors; D013449:Sulfonamides; D014157:Transcription Factors; D014404:Tubulin; D014746:Vimentin; D007633:Keratins; D019810:Sodium Azide; D019833:Genistein