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

XIAOMEI ZHANG; SILVINA BOCCA; ANAHI FRANCHI; SANDRA ANDERSON; MANDEEP KAUR; VLADIMIR B BAJIC; SERGIO OEHNINGER

MOLECULAR HUMAN REPRODUCTION.

OXFORD UNIV PRESS

Año: 2010 vol. 16 p. 347 - 360

1360-9947

We examined whether 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 absence and presence of E2 and P4, and cAMP) on mRNA expression of genes modulated during the window of implantation in GnRH analogues/rFSH-treated ART 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 presence of putative steroid response elements (EREs and PREs). Results: Ishikawa cells were cytokeratin+/vimentin-, and expressed ERα, ERβ, PR and GnRH-R proteins. At 6 and 24 hours, neither LA nor GA alone had an effect on gene expression. GnRH analogues alone or following E2 and/or P4 co-incubation for 24 hours 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.