title: Regulatory effects of estetrol on the endothelial plasminogen pathway and endothelial cell migration.
abstract: BACKGROUND
  Estetrol (E4) is a natural estrogen produced solely during human pregnancy. E4 is suitable for clinical use since it acts as a selective estrogen receptor modulator. In clinical trials E4 has been seen to have little or no effect on coagulation. Hence, it is interesting to investigate whether E4 alters endothelial-dependent fibrinolysis.
  
  
  OBJECTIVE
  We studied the effects of E4 on the fibrinolytic system and whether this could influence the ability of endothelial cells to migrate. In addition, we compared the effects of E4 with those of 17β-estradiol (E2).
  
  
  METHODS
  Human umbilical vein endothelial cells (HUVEC) were obtained from healthy women. Expression of plasminogen-activator inhibitor-1 (PAI-1), urokinase-type plasminogen activator (u-PA) and tissue plasminogen activator (t-PA) proteins was evaluated by Western blot analysis. Endothelial cell migration was studied by razor-scrape horizontal and multiwell insert systems assays.
  
  
  RESULTS
  E4 increased the expression of t-PA, u-PA and PAI-1 in HUVEC, but less so than did equimolar amounts of E2. The effects of E4 on t-PA, u-PA and PAI-1 were mediated by the induction of the early-immediate genes c-Jun and c-Fos. E4 in combination with E2 antagonized the effects induced by pregnancy-like E2 concentrations but did not impair the effects of postmenopausal-like E2 levels. We also found that the increased synthesis of PAI-1, u-PA and t-PA induced by E2 and E4 is important for horizontal and three-dimensional migration of HUVEC.
  
  
  CONCLUSIONS
  These results support the hypothesis that E4 acts as an endogenous selective estrogen receptor modulator (SERM), controlling the fibrinolytic system and endothelial cell migration.
authors.affiliation: Molecular and Cellular Gynecological Endocrinology Laboratory (MCGEL), Department of Clinical and Experimental Medicine, University of Pisa, Pisa, PI, Italy.
authors.forename: Maria Magdalena
authors.initials: MM
authors.lastname: Montt-Guevara
authors.affiliation: Molecular and Cellular Gynecological Endocrinology Laboratory (MCGEL), Department of Clinical and Experimental Medicine, University of Pisa, Pisa, PI, Italy.
authors.forename: Giulia
authors.initials: G
authors.lastname: Palla
authors.affiliation: Molecular and Cellular Gynecological Endocrinology Laboratory (MCGEL), Department of Clinical and Experimental Medicine, University of Pisa, Pisa, PI, Italy.
authors.forename: Stefania
authors.initials: S
authors.lastname: Spina
authors.affiliation: Molecular and Cellular Gynecological Endocrinology Laboratory (MCGEL), Department of Clinical and Experimental Medicine, University of Pisa, Pisa, PI, Italy.
authors.forename: Guja
authors.initials: G
authors.lastname: Bernacchi
authors.affiliation: Molecular and Cellular Gynecological Endocrinology Laboratory (MCGEL), Department of Clinical and Experimental Medicine, University of Pisa, Pisa, PI, Italy.
authors.forename: Elena
authors.initials: E
authors.lastname: Cecchi
authors.affiliation: Molecular and Cellular Gynecological Endocrinology Laboratory (MCGEL), Department of Clinical and Experimental Medicine, University of Pisa, Pisa, PI, Italy.
authors.forename: Adrian Esteban
authors.initials: AE
authors.lastname: Campelo
authors.affiliation: Molecular and Cellular Gynecological Endocrinology Laboratory (MCGEL), Department of Clinical and Experimental Medicine, University of Pisa, Pisa, PI, Italy.
authors.forename: Jorge Eduardo
authors.initials: JE
authors.lastname: Shortrede
authors.affiliation: Molecular and Cellular Gynecological Endocrinology Laboratory (MCGEL), Department of Clinical and Experimental Medicine, University of Pisa, Pisa, PI, Italy.
authors.forename: Alessio
authors.initials: A
authors.lastname: Canu
authors.affiliation: Molecular and Cellular Gynecological Endocrinology Laboratory (MCGEL), Department of Clinical and Experimental Medicine, University of Pisa, Pisa, PI, Italy. Electronic address: tommaso.simoncini@med.unipi.it.
authors.forename: Tommaso
authors.initials: T
authors.lastname: Simoncini
mesh_terms: D015153:Blotting, Western; D002465:Cell Movement / Q000187:drug effects*; D002478:Cells, Cultured; D042783:Endothelial Cells; D004730:Endothelium, Vascular / Q000187:drug effects; D004953:Estetrol / Q000494:pharmacology*; D004958:Estradiol / Q000494:pharmacology; D004967:Estrogens / Q000494:pharmacology; D005260:Female; D005342:Fibrinolysis / Q000187:drug effects*; D061307:Human Umbilical Vein Endothelial Cells / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D017395:Plasminogen Activator Inhibitor 1 / Q000187:drug effects* / Q000378:metabolism; D020845:Selective Estrogen Receptor Modulators / Q000494:pharmacology*; D010959:Tissue Plasminogen Activator / Q000187:drug effects* / Q000378:metabolism; D014568:Urokinase-Type Plasminogen Activator / Q000187:drug effects* / Q000378:metabolism
chemical_list: D004967:Estrogens; D017395:Plasminogen Activator Inhibitor 1; C051321:SERPINE1 protein, human; D020845:Selective Estrogen Receptor Modulators; D004958:Estradiol; D010959:Tissue Plasminogen Activator; D014568:Urokinase-Type Plasminogen Activator; D004953:Estetrol
keywords: Estetrol; Estrogen; HUVEC; PAI-1; t-PA; u-PA