Effect of GnRH analogs on cell proliferation, apoptosis and expression of Bcl-2, Bax, Fas and FasL in epithelial endometrial cultures from patients with endometriosis.

BILOTAS M; BARAÑAO RI; BUQUET R.; SUELDO C.; TESONE M.; MERESMAN GF

HUMAN REPRODUCTION

Año: 2007 vol. 22 p. 644 - 653

0268-1161

BACKGROUND: Our purpose was to evaluate the effect of the GnRH agonist, Leuprolide Acetate (LA), and the GnRH antagonist, Antide, on apoptosis and apoptosis-related proteins expression in endometrial epithelial cultures (EEC) from patients with endometriosis and controls. 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 assesed by the acridine orange-ethidium bromide technique and protein expression was evaluated by Western blot and immunocytochemistry.  RESULTS: LA 100 ng/ml and 1000 ng/ml  increased the %ApC in EEC from patients with endometriosis (p<0.05) and controls (p<0.05; p<0.01). Antide 10-5M increased the %ApC in EEC from patients with endometriosis and controls (p<0.01). Bax expression increased after treatment with LA, Antide and LA+Antide (p<0.05; p<0.001; p<0.001). Bcl-2 expression decreased after exposure to LA and Antide (p<0.001; p<0.01). FasL expression incresed after LA, Antide and LA+Antide treatments (p<0.01; p<0.001; p<0.01). No significant changes were observed on Fas expression. CONCLUSIONS: GnRH analogues enhance apoptosis by increasing the expression of pro-apoptotic proteins Bax and FasL and diminishing the expression of the anti-apoptotic protein Bcl-2.