title: Estrogen metabolites in the release of inflammatory mediators from human amnion-derived cells.
abstract: OBJECTIVE
  Human amnion-derived cells have been used as in vitro models to test the release of inflammatory mediators, such as arachidonic acid (AA) and prostaglandin E(2) (PGE(2)). We compared estrogen metabolites for their ability to induce AA release, to influence PGE(2) production and to interact toward intracellular estrogen receptors (ERs).
  
  
  METHODS
  Metabolite effects on AA and PGE(2) release were examined by radiolabelled substrate incorporation and by colorimetric enzyme immunoassays, respectively. [(3)H]17-β-estradiol binding displacements were performed on Ro-20-1724 treated whole cells.
  
  
  RESULTS
  In WISH cells, estrone, 2-hydroxy-estrone and estriol induced a rapid dose dependent release of AA that was not inhibited by cycloheximide. Estrone and 2-hydroxy-estrone showed biphasic dose-response curves of PGE(2), whereas estriol and 16-α-hydroxy-estrone increased PGE(2) levels at high concentrations. 2-methoxy-estrone, 4-hydroxy-estradiol and 4-hydroxy-estrone did not significantly affect PGE(2) release. 2-methoxy-estradiol and 2-hydroxy-estradiol decreased the PGE(2) release. Effects of metabolites on PGE(2) were inhibited by cycloheximide and by the ER antagonist tamoxifen. In AV3 cells PGE(2) production was poorly detectable. On Ro-20-1724 treated WISH cells the K(i) of 17-β-estradiol was 29.2 ± 5.4 nM. Estrone, 2-methoxy-estrone and 2-methoxy-estradiol showed similar affinity values. The hydroxyl substituent at position 2, 4 and 16 decreased or markedly increased the affinity for estradiol or estrone derivatives, respectively.
  
  
  CONCLUSIONS
  The estrogen metabolites induced nongenomic effects on AA release from WISH cells. The influence on PGE(2) release was detectable only on WISH cells. These effects appeared genomic and mediated by intracellular ERs, whose properties seemed strongly dependent on intracellular cAMP levels.
authors.affiliation: Department of Biology, General Physiology sect., University of Ferrara, via L. Borsari, 46 - 44121 Ferrara, Italy.
authors.forename: Barbara
authors.initials: B
authors.lastname: Pavan
authors.forename: Guglielmo
authors.initials: G
authors.lastname: Paganetto
authors.forename: Alessandro
authors.initials: A
authors.lastname: Dalpiaz
authors.forename: Carla
authors.initials: C
authors.lastname: Biondi
authors.forename: Laura
authors.initials: L
authors.lastname: Lunghi
mesh_terms: D000650:Amnion / Q000187:drug effects* / Q000378:metabolism; D016718:Arachidonic Acid / Q000378:metabolism*; D001667:Binding, Competitive; D018929:Cell Culture Techniques; D002460:Cell Line; D015232:Dinoprostone / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004847:Epithelial Cells / Q000187:drug effects* / Q000378:metabolism; D045166:Estradiol Congeners / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology*; D006801:Humans; D018836:Inflammation Mediators / Q000378:metabolism*; D015394:Molecular Structure
chemical_list: D045166:Estradiol Congeners; D018836:Inflammation Mediators; D016718:Arachidonic Acid; D015232:Dinoprostone