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Localization of Interferon Regulatory Factor-1 (IRF-1) in Nonpregnant Human Endometrium: Expression of IRF-1 Is Up-Regulated by Prolactin during the Secretory Phase of the Menstrual Cycle

Medical Research Council Reproductive Biology Unit (H.N.J., S.C.B.) and Obstetrics and Gynecology (H.O.D.C.), University of Edinburgh Center for Reproductive Biology, Edinburgh, United Kingdom EH3 9ET; and the Department of Medicine (L.-y.Y.-L.), Baylor College of Medicine, Houston, Texas 77030

Address all correspondence and requests for reprints to: Dr. H. N. Jabbour, Medical Research Council Reproductive Biology Unit, 37 Chalmers Street, Edinburgh, United Kingdom EH3 9ET. E-mail: h.jabbour{at}ed-rbu.mrc.ac.uk

PRL expression in the human uterus is up-regulated during the mid to late secretory phase of the menstrual cycle. This coincides with up-regulation of the expression of the PRL receptor, which is localized primarily to the endometrial glandular epithelial cells. Recent data have demonstrated activation of the Jak (Janus kinase)/Stat (signal transducer and activator of transcription) signaling pathway in the secretory endometrium after stimulation with exogenous PRL. However, the target genes for the action of PRL on the endometrial epithelial cells have not been elucidated. In this study we have investigated the pattern/site of expression of the transcription factor interferon regulatory factor-1 (IRF-1) as well as the effect of exogenous PRL on the transcription of IRF-1 in the human endometrium during the mid to late secretory phase of the menstrual cycle. Expression of the IRF-1 gene was confirmed by RNase protection assays using a 260-bp homologous [-³²P]UTP-labeled IRF-1 complementary ribonucleic acid (RNA) probe and 10 µg total RNA extracted from human endometrium (n = 5) collected between days 19 and 26 of the menstrual cycle. Northern and Western blot analyses were conducted on secretory phase human endometrium (n = 3) using human [-³²P]dCTP-labeled IRF-1 complementary DNA and antihuman IRF-1 antibody. Expression of the IRF-1 gene in the secretory phase endometrium was encoded by a RNA transcript of approximately 2.1 kb and a protein of 48 kDa. Furthermore, expression of the IRF-1 gene in the secretory phase endometrium was localized by immunohistochemistry predominantly to the glandular epithelial cells as has been shown previously for the PRL receptor. To investigate the effect of PRL on expression of IRF-1, human endometrial biopsies (n = 3) collected between days 24–26 of the menstrual cycle were cultured in the presence of cycloheximide with or without 100 ng/mL human PRL for 2 and 4 h. Culture of endometrial tissue with PRL for 2 and 4 h resulted in 2.9 ± 0.3-fold (P < 0.01) and 1.7 ± 0.1-fold induction of expression of the IRF-1 gene, respectively. These data demonstrate the expression of the transcription factor IRF-1 in the glandular epithelium of the endometrium and its regulation by PRL during the secretory phase of the menstrual cycle. Previous observations of the temporal up-regulation of expression of both PRL and PRL receptors in the secretory human endometrium and their localization to the stromal and glandular compartments, respectively, suggest that endometrial PRL mediates transcription of the IRF-1 gene in a paracrine fashion.

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