Do GnRH analogues directly affect human endometrial epithelial cell gene expression?

• Published in: Molecular human reproduction 2010-05, Vol.16 (5), p.347-360
• Main Authors: Zhang, Xiaomei, Bocca, Silvina, Franchi, Anahi, Anderson, Sandra, Kaur, Mandeep, Bajic, Vladimir B., Oehninger, Sergio
• Format: Article
• Language: English
• Subjects: Analysis of Variance; Biological and medical sciences; Carrier Proteins - genetics; Cells, Cultured; Embryology: invertebrates and vertebrates. Teratology; endometrium; Endometrium - cytology; Endometrium - drug effects; Endosomal Sorting Complexes Required for Transport - genetics; Epithelial Cells - cytology; Epithelial Cells - drug effects; EREs/PREs identification; Estradiol - pharmacology; Estrogens - pharmacology; Female; Fertility Agents, Female - pharmacology; Fundamental and applied biological sciences. Psychology; gene expression; Gene Expression - drug effects; GnRH; Gonadotropin-Releasing Hormone - analogs & derivatives; Gonadotropin-Releasing Hormone - pharmacology; Hormone Antagonists - pharmacology; Humans; Immunohistochemistry; Insulin-Like Growth Factor Binding Protein 5 - genetics; Ishikawa cells; Leuprolide - pharmacology; Progesterone - pharmacology; Progestins - pharmacology; Response Elements - drug effects; Response Elements - genetics; Reverse Transcriptase Polymerase Chain Reaction; Saposins - genetics; Sorting Nexins; Transcription Factor TFIIIA - genetics; Vesicular Transport Proteins - genetics
• ISSN: 1360-9947; 1460-2407
• DOI: 10.1093/molehr/gaq012

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 E2 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 ERα, ERβ, 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 E2 and/or P4 co-incubation for 24 h also had no effect on gene expression, but P4 significantly increased expression of CHMP1A. E2 + P4 treatment for 4 days, alone or followed by GA, had no effect, but E2 + 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.