Involvement of nuclear factor-kappaB in macrophage migration inhibitory factor gene transcription up-regulation induced by interleukin- 1 beta in ectopic endometrial cells.
OBJECTIVE
To investigate the involvement of the nuclear factor (NF)-kappaB in the interleukin (IL)-1 beta-mediated macrophage migration inhibitory factor (MIF) gene activation.


METHODS
Prospective study.


METHODS
Human reproduction research laboratory.


METHODS
Nine women with endometriotic lesions.


METHODS
Endometriotic lesions were obtained during laparoscopic surgery.


METHODS
The MIF protein secretion was analyzed by ELISA, MIF mRNA expression by quantitative real-time polymerase chain reaction (PCR), NF-kappaB translocation into the nucleus by electrophoresis mobility shift assay, I kappaB phosphorylation and degradation by Western blot, and human MIF promoter activity by transient cell transfection.


RESULTS
This study showed a significant dose-dependent increase of MIF protein secretion and mRNA expression, the NF-kappaB translocation into the nucleus, I kappaB phosphorylation, I kappaB degradation, and human MIF promoter activity in endometriotic stromal cells in response to IL-1 beta. Curcumin (NF-kappaB inhibitor) significantly inhibited all these IL-1 beta-mediated effects. Analysis of the activity of deletion constructs of the human MIF promoter and a computer search localized two putative regulatory elements corresponding to NF-kappaB binding sites at positions -2538/-2528 bp and -1389/-1380 bp.


CONCLUSIONS
This study suggests the involvement of the nuclear transcription factor NF-kappaB in MIF gene activation in ectopic endometrial cells in response to IL-1 beta and identifies a possible pathway of endometriosis-associated inflammation and ectopic cell growth.
Veillat
Véronique
V
Unité d'Endocrinologie de la Reproduction, Centre de Recherche, Hôpital Saint-François d'Assise, Centre Hospitalier Universitaire de Québec, Faculté de Médecine, Université Laval, Québec, Canada.
Lavoie
Catherine Herrmann
CH
Metz
Christine N
CN
Roger
Thierry
T
Labelle
Yves
Y
Akoum
Ali
A
D018929:Cell Culture Techniques; D004715:Endometriosis / Q000235:genetics* / Q000473:pathology; D005260:Female; D006801:Humans; D053583:Interleukin-1beta / Q000494:pharmacology*; D008263:Macrophage Migration-Inhibitory Factors / Q000235:genetics*; D016328:NF-kappa B / Q000502:physiology*; D011247:Pregnancy; D011271:Pregnancy, Ectopic / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics; D014158:Transcription, Genetic / Q000187:drug effects*; D015854:Up-Regulation / Q000187:drug effects*
D053583:Interleukin-1beta; D008263:Macrophage Migration-Inhibitory Factors; D016328:NF-kappa B; D012333:RNA, Messenger