title: Do GnRH analogues directly affect human endometrial epithelial cell gene expression?
abstract: We examined whether Gonadotrophin-releasing hormone (GnRH) analogues [leuprolide acetate (LA) and ganirelix acetate (GA)] modulate gene expression in Ishikawa cells used as surrogate for human endometrial epithelial cells in vitro. The specific aims were: (i) to study the modulatory effect of GnRH analogues by RT-PCR [in the absence and presence of E(2) and P4, and cyclic adenosine monophosphate (cAMP)] on mRNA expression of genes modulated during the window of implantation in GnRH analogues/rFSH-treated assisted reproductive technology cycles including OPTINEURIN (OPTN), CHROMATIN MODIFYING PROTEIN (CHMP1A), PROSAPOSIN (PSAP), IGFBP-5 and SORTING NEXIN 7 (SNX7), and (ii) to analyze the 5'-flanking regions of such genes for the presence of putative steroid-response elements [estrogen-response elements (EREs) and P4-response element (PREs)]. Ishikawa cells were cytokeratin+/vimentin- and expressed ERalpha, ERbeta, PR and GnRH-R proteins. At 6 and 24 h, neither LA nor GA alone had an effect on gene expression. GnRH analogues alone or following E(2) and/or P4 co-incubation for 24 h also had no effect on gene expression, but P4 significantly increased expression of CHMP1A. E(2) + P4 treatment for 4 days, alone or followed by GA, had no effect, but E(2) + P4 treatment followed by LA significantly decreased IGFBP-5 expression. The addition of 8-Br cAMP did not modify gene expression, with the exception of IGFBP-5 that was significantly increased. The GnRH analogues did not modify intracellular cAMP levels. We identified conserved EREs for OPN, CHMP1A, SNX7 and PSAP and PREs for SNX7. We conclude that GnRH analogues appear not to have major direct effects on gene expression of human endometrial epithelial cells in vitro.
authors.affiliation: Department of Obstetrics and Gynecology, The Jones Institute for Reproductive Medicine, Eastern Virginia Medical School, Norfolk, VA 23507, USA.
authors.forename: Xiaomei
authors.initials: X
authors.lastname: Zhang
authors.forename: Silvina
authors.initials: S
authors.lastname: Bocca
authors.forename: Anahi
authors.initials: A
authors.lastname: Franchi
authors.forename: Sandra
authors.initials: S
authors.lastname: Anderson
authors.forename: Mandeep
authors.initials: M
authors.lastname: Kaur
authors.forename: Vladimir B
authors.initials: VB
authors.lastname: Bajic
authors.forename: Sergio
authors.initials: S
authors.lastname: Oehninger
mesh_terms: D000704:Analysis of Variance; D002352:Carrier Proteins / Q000235:genetics; D018797:Cell Cycle Proteins; D002478:Cells, Cultured; D004717:Endometrium / Q000166:cytology / Q000187:drug effects*; D056827:Endosomal Sorting Complexes Required for Transport / Q000235:genetics; D004847:Epithelial Cells / Q000166:cytology / Q000187:drug effects*; D004958:Estradiol / Q000494:pharmacology; D004967:Estrogens / Q000494:pharmacology; D005260:Female; D005300:Fertility Agents, Female / Q000494:pharmacology; D015870:Gene Expression / Q000187:drug effects*; D007987:Gonadotropin-Releasing Hormone / Q000031:analogs & derivatives* / Q000494:pharmacology; D006727:Hormone Antagonists / Q000494:pharmacology; D006801:Humans; D007150:Immunohistochemistry; D018975:Insulin-Like Growth Factor Binding Protein 5 / Q000235:genetics; D016729:Leuprolide / Q000494:pharmacology*; D026901:Membrane Transport Proteins; D011374:Progesterone / Q000494:pharmacology; D011372:Progestins / Q000494:pharmacology; D020218:Response Elements / Q000187:drug effects / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D049231:Saposins / Q000235:genetics; D058305:Sorting Nexins; D035582:Transcription Factor TFIIIA / Q000235:genetics; D033921:Vesicular Transport Proteins / Q000235:genetics
chemical_list: C443709:CHMP1A protein, human; D002352:Carrier Proteins; D018797:Cell Cycle Proteins; D056827:Endosomal Sorting Complexes Required for Transport; D004967:Estrogens; D005300:Fertility Agents, Female; D006727:Hormone Antagonists; D018975:Insulin-Like Growth Factor Binding Protein 5; D026901:Membrane Transport Proteins; C409460:OPTN protein, human; C483345:PSAP protein, human; D011372:Progestins; D049231:Saposins; D058305:Sorting Nexins; D035582:Transcription Factor TFIIIA; D033921:Vesicular Transport Proteins; D007987:Gonadotropin-Releasing Hormone; D011374:Progesterone; D004958:Estradiol; D016729:Leuprolide; C061018:ganirelix