title: Gonadotropin releasing hormone antagonists suppress aromatase and anti-Müllerian hormone expression in human granulosa cells.
abstract: OBJECTIVE
  To investigate the effects of a gonadotropin-releasing hormone antagonist (GnRH-ANT) on the expression of anti-Müllerian Hormone (AMH) and aromatase (via the exon CYP19IIa promoter), in cultured human granulosa cells (hGCs) and the human granulosa cell line (HGL5).
  
  
  METHODS
  Primary cell cultures of hGCs and culture of HGL5 cells.
  
  
  METHODS
  Academic center.
  
  
  METHODS
  Women undergoing IVF because of male factor, tubal infertility, or donor eggs.
  
  
  METHODS
  hGCs and HGL5 cells were treated with a GnRH-ANT (1 nM and 1 μM) alone or in combination with cAMP (1 mM). Media was collected and stored at -80°C until assayed.
  
  
  METHODS
  mRNA levels of CYP19 IIa, AMH, steroidogenic factor 1 (SF-1) and liver receptor homologue-1 (LRH-1) were determined by quantitative polymerase chain reaction. ELISA was used to determined estradiol (E(2)) levels in the culture media. Pooled results from triplicate experiments were analyzed using one-way analysis of variance with Student-Newman-Keuls multiple-comparison methods.
  
  
  RESULTS
  The GnRH-ANT decreased the expressions of CYP19 IIa, AMH, SF-1, and LRH-1. cAMP induced aromatase and AMH expression. Cotreatment with cAMP and GnRH-ANT caused a dose-dependent suppression of AMH and CYP19 IIa mRNA. A GnRH agonist (GnRH-A) increased the mRNA expressions of CYP 19 IIa and AMH. The GnRH-ANT decreased E(2) production in cultured hGCs.
  
  
  CONCLUSIONS
  GnRH-ANTs, in addition to their central suppressive effects on the pituitary, may have a direct effect on ovarian granulosa cells with inhibition of aromatase and AMH expression. Furthermore, the inhibitory effect could be mediated via suppression of SF-1 and LRH-1, and may play a role in estrogen-mediated ovarian folliculogenesis.
authors.affiliation: Department of Obstetrics and Gynecology, the University of Texas Southwestern Medical Center, Dallas, Texas, USA.
authors.forename: Nurit
authors.initials: N
authors.lastname: Winkler
authors.forename: Orhan
authors.initials: O
authors.lastname: Bukulmez
authors.forename: Daniel B
authors.initials: DB
authors.lastname: Hardy
authors.forename: Bruce R
authors.initials: BR
authors.lastname: Carr
mesh_terms: D000328:Adult; D054304:Anti-Mullerian Hormone / Q000378:metabolism*; D001141:Aromatase / Q000378:metabolism*; D002460:Cell Line; D002478:Cells, Cultured; D000242:Cyclic AMP / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004958:Estradiol / Q000378:metabolism; D005260:Female; D007987:Gonadotropin-Releasing Hormone / Q000819:agonists / Q000031:analogs & derivatives* / Q000037:antagonists & inhibitors* / Q000494:pharmacology; D006107:Granulosa Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism*; D006801:Humans; D016729:Leuprolide / Q000494:pharmacology; D012333:RNA, Messenger / Q000378:metabolism; D018160:Receptors, Cytoplasmic and Nuclear / Q000378:metabolism; D054339:Steroidogenic Factor 1 / Q000378:metabolism
chemical_list: C082631:NR5A2 protein, human; D012333:RNA, Messenger; D018160:Receptors, Cytoplasmic and Nuclear; D054339:Steroidogenic Factor 1; D007987:Gonadotropin-Releasing Hormone; D004958:Estradiol; D054304:Anti-Mullerian Hormone; D000242:Cyclic AMP; D001141:Aromatase; D016729:Leuprolide; C062876:cetrorelix