title: MicroRNA miR-145 inhibits proliferation, invasiveness, and stem cell phenotype of an in vitro endometriosis model by targeting multiple cytoskeletal elements and pluripotency factors.
abstract: OBJECTIVE
  To study the function of miR-145, known to be dysregulated in endometriosis, and to identify its target genes in an in vitro endometriosis model.
  
  
  METHODS
  Experimental laboratory study.
  
  
  METHODS
  University medical centers.
  
  
  METHODS
  Primary endometrial stroma cells were derived from eutopic endometrium of three American Society for Reproductive Medicine stage III endometriosis patients and from ectopic lesions of four patients with deep infiltrating endometriosis.
  
  
  METHODS
  The human endometriotic cell line 12Z and primary eutopic and ectopic endometrial stroma cells were transiently transfected with miR-145 precursors or anti-miR-145 inhibitors and investigated for posttranscriptional regulation of predicted target genes and changes in cell behavior.
  
  
  METHODS
  Predicted target expression was measured by quantitative reverse transcription-polymerase chain reaction and Western blotting, and altered cell behavior was monitored by cell proliferation assays. The 12Z cells were additionally investigated by Matrigel invasion assays, cell cycle analysis, side population analysis, and aldehyde dehydrogenase activity assays.
  
  
  RESULTS
  In all cells investigated, miR-145 overexpression inhibited cell proliferation and induced down-regulation of FASCIN-1, SOX2, and MSI2. In 12Z cells miR-145 upregulation increased Matrigel invasiveness and reduced side population and aldehyde dehydrogenase-1 activity. Additional down-regulated targets in 12Z cells included OCT4, KLF4, PODXL, JAM-A, and SERPINE1/PAI-1. ACTG2 and TAGLN were up-regulated upon pre-miR-145 transfection. JAM-A, FASCIN-1, and PAI-I down-regulation in 12Z cells were confirmed by Western blotting.
  
  
  CONCLUSIONS
  miR-145 inhibits endometriotic cell proliferation, invasiveness, and stemness by targeting multiple pluripotency factors, cytoskeletal elements, and protease inhibitors.
authors: Department of Gynecology and Obstetrics, Münster University Hospital, Münster, Germany.
authors: Marlene
authors: M
authors: Adammek
authors: Burkhard
authors: B
authors: Greve
authors: Nadja
authors: N
authors: Kässens
authors: Cornelia
authors: C
authors: Schneider
authors: Kathrin
authors: K
authors: Brüggemann
authors: Andreas N
authors: AN
authors: Schüring
authors: Anna
authors: A
authors: Starzinski-Powitz
authors: Ludwig
authors: L
authors: Kiesel
authors: Martin
authors: M
authors: Götte
mesh_terms: D000328:Adult; D002352:Carrier Proteins / Q000235:genetics; D015815:Cell Adhesion Molecules / Q000235:genetics; D002460:Cell Line; D049109:Cell Proliferation; D003599:Cytoskeleton / Q000502:physiology*; D004715:Endometriosis / Q000235:genetics* / Q000473:pathology*; D005260:Female; D006801:Humans; D066298:In Vitro Techniques; D000090062:Kruppel-Like Factor 4; D035683:MicroRNAs / Q000235:genetics*; D008840:Microfilament Proteins / Q000235:genetics; D010641:Phenotype; D017395:Plasminogen Activator Inhibitor 1 / Q000235:genetics; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000502:physiology; D012323:RNA Processing, Post-Transcriptional / Q000502:physiology; D016601:RNA-Binding Proteins / Q000235:genetics; D011956:Receptors, Cell Surface / Q000235:genetics; D055748:SOXB1 Transcription Factors / Q000235:genetics; D012720:Severity of Illness Index; D017154:Stromal Cells / Q000166:cytology / Q000502:physiology
chemical_list: D002352:Carrier Proteins; D015815:Cell Adhesion Molecules; C407243:F11R protein, human; C502658:FSCN1 protein, human; C000715467:KLF4 protein, human; D000090062:Kruppel-Like Factor 4; C541010:MIRN145 microRNA, human; C438151:MSI2 protein, human; D035683:MicroRNAs; D008840:Microfilament Proteins; D017395:Plasminogen Activator Inhibitor 1; D016601:RNA-Binding Proteins; D011956:Receptors, Cell Surface; C051321:SERPINE1 protein, human; C527782:SOX2 protein, human; D055748:SOXB1 Transcription Factors