Hum. Reprod. Advance Access originally published online on November 8, 2006
Human Reproduction 2007 22(3):644-653; doi:10.1093/humrep/del423
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Effect of GnRH analogues on apoptosis and expression of Bcl-2, Bax, Fas and FasL proteins in endometrial epithelial cell cultures from patients with endometriosis and controls
1 Instituto de Biología y Medicina Experimental (IBYME) 2 Departamento de Ginecología, Hospital de Clínicas ‘José de San Martín’ and 3 Centro de Estudios en Ginecología y Reproducción (CEGyR), Buenos Aires, Argentina
4 To whom correspondence should be addressed at: Instituto de Biología y Medicina Experimental (IBYME), C1428ADN Buenos Aires, Argentina. E-mail: mabilotas{at}dna.uba.ar
BACKGROUND: Our purpose was to evaluate the effect of the GnRH agonist (GnRHa), leuprolide acetate (LA), and the GnRH antagonist (GnRHant), Antide, on apoptosis and expression of apoptosis-related proteins in endometrial epithelial cell (EEC) cultures from patients with endometriosis and controls (infertile women without endometriosis). METHODS: Biopsy specimens of eutopic endometrium were obtained from 22 patients with endometriosis and from 14 women that served as controls. Apoptosis was examined in EEC after incubation with LA and Antide. Bax, Bcl-2, Fas and FasL expression was evaluated after exposure to LA, Antide or a combination of both. The percentage of apoptotic cells (%ApC) was assessed by the acridine orange–ethidium bromide technique, and protein expression was evaluated by western blot and immunocytochemistry. RESULTS: LA 100 and 1000 ng/ml increased the %ApC in EEC from patients with endometriosis (both P < 0.05) and controls (p < 0.05 and P < 0.01, respectively). Antide 10–5 M increased the %ApC in EEC from patients with endometriosis and controls (P < 0.01). In EEC from women with endometriosis, Bax expression increased after treatment with LA, Antide and LA + Antide (P < 0.05, P < 0.001 and P < 0.001), whereas Bcl-2 expression decreased after exposure to LA and Antide (P < 0.001 and P < 0.01). FasL expression increased after LA, Antide and LA + Antide treatments (P < 0.01, P < 0.001 and P < 0.01). No significant changes were observed on Fas expression. CONCLUSIONS: GnRH analogues enhanced apoptosis in EEC, and this was accompanied by an increase in expression of the pro-apoptotic proteins Bax and FasL and a decrease in expression of the anti-apoptotic protein Bcl-2.
Key words: apoptosis/endometriosis/GnRH analogues
Submitted on March 28, 2006; resubmitted on August 16, 2006; resubmitted on September 8, 2006; accepted on September 18, 2006.
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