This Article Full Text Full Text (PDF) All Versions of this Article: 145/3/1248    most recent Author Manuscript (PDF) Purchase Article View Shopping Cart Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Copyright Permission Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Soloff, M. S. Articles by Anderson, G. D. Search for Related Content PubMed PubMed Citation Articles by Soloff, M. S. Articles by Anderson, G. D.

In Situ Analysis of Interleukin-1-Induced Transcription of cox-2 and il-8 in Cultured Human Myometrial Cells

Department of Obstetrics & Gynecology (M.S.S., D.L.C., Y.-J.J., G.D.A.), Sealy Center for Molecular Science (M.S.S.), University of Texas Medical Branch, Galveston, Texas 77555-1062

Address all correspondence and requests for reprints to: Melvyn S. Soloff, Department of Obstetrics and Gynecology, University of Texas Medical Branch, 301 University Boulevard, Galveston, Texas 77555-1062. E-mail: msoloff{at}utmb.edu.

The specific binding of transcription factors to DNA has been shown to be inhibited by chromatin structure and increased by cooperative interactions with other proteins. Consequently, in situ analysis using chromatin immunoprecipitation offers the most accurate view of transcriptional control. Transient transfection studies and in vitro analyses of IL-1-induced cox-2 transcription in a number of cell types have indicated regulation by either nuclear factor B (NF-B) or CCAAT/enhancer binding protein (C/EBPß), or both acting cooperatively. To determine the mechanisms of COX-2 (cyclooxygenase or prostaglandin endoperoxide synthase) induction in cultured human myometrial cells in situ, we examined the cross-linking of the RelA subunit of NF-B and C/EBPß to the cox-2 promoter and flanking sequences. As a control, we inspected the interaction of these transcription factors with the IL-8 gene, which has been shown in other cell types to be activated by the cooperative interaction of NF-B and C/EBPß. Indeed, both transcription factors were cross-linked to the il-8 promoter after IL-1 treatment, but only RelA was cross-linked to cox-2 DNA. The il-8 promoter was also found to physically interact with proteins cross-linked to sites further upstream. IL-1 treatment also increased polymerase II cross-linking to both promoters and increased histone H4 acetylation at specific sites. These results indicate that modification of chromatin structure is part of the response to IL-1 stimulation. Chromatin immunoprecipitation thus provides critical insight into the mechanisms of COX-2 and IL-8 expression in human myometrial cells.

This article has been cited by other articles:

				V. Bezzerri, M. Borgatti, E. Nicolis, I. Lampronti, M. C. Dechecchi, I. Mancini, P. Rizzotti, R. Gambari, and G. Cabrini Transcription Factor Oligodeoxynucleotides to NF-{kappa}B Inhibit Transcription of IL-8 in Bronchial Cells Am. J. Respir. Cell Mol. Biol., July 1, 2008; 39(1): 86 - 96. [Abstract] [Full Text] [PDF]

				Z. Liang, S.R. Sooranna, N. Engineer, M. Tattersall, S. Khanjani, P.R. Bennett, L. Myatt, and M.R. Johnson Prostaglandin F2-alpha receptor regulation in human uterine myocytes Mol. Hum. Reprod., April 1, 2008; 14(4): 215 - 223. [Abstract] [Full Text] [PDF]

				W. E Ackerman IV, T. L.S Summerfield, D. D Vandre, J. M Robinson, and D. A Kniss Nuclear Factor-Kappa B Regulates Inducible Prostaglandin E Synthase Expression in Human Amnion Mesenchymal Cells Biol Reprod, January 1, 2008; 78(1): 68 - 76. [Abstract] [Full Text] [PDF]

				G. Chevillard, A. Derjuga, D. Devost, H. H Zingg, and V. Blank Identification of interleukin-1{beta} regulated genes in uterine smooth muscle cells Reproduction, December 1, 2007; 134(6): 811 - 822. [Abstract] [Full Text] [PDF]

				D. Markovic, M. Vatish, M. Gu, D. Slater, R. Newton, H. Lehnert, and D. K. Grammatopoulos The Onset of Labor Alters Corticotropin-Releasing Hormone Type 1 Receptor Variant Expression in Human Myometrium: Putative Role of Interleukin-1{beta} Endocrinology, July 1, 2007; 148(7): 3205 - 3213. [Abstract] [Full Text] [PDF]

				A. R. Mohan, S. R. Sooranna, T. M. Lindstrom, M. R. Johnson, and P. R. Bennett The Effect of Mechanical Stretch on Cyclooxygenase Type 2 Expression and Activator Protein-1 and Nuclear Factor-{kappa}B Activity in Human Amnion Cells Endocrinology, April 1, 2007; 148(4): 1850 - 1857. [Abstract] [Full Text] [PDF]

				D. B. Hardy, B. A. Janowski, D. R. Corey, and C. R. Mendelson Progesterone Receptor Plays a Major Antiinflammatory Role in Human Myometrial Cells by Antagonism of Nuclear Factor-{kappa}B Activation of Cyclooxygenase 2 Expression Mol. Endocrinol., November 1, 2006; 20(11): 2724 - 2733. [Abstract] [Full Text] [PDF]

				D. B. Zaragoza, R. R. Wilson, B. F. Mitchell, and D. M. Olson The Interleukin 1beta-Induced Expression of Human Prostaglandin F2alpha Receptor Messenger RNA in Human Myometrial-Derived ULTR Cells Requires the Transcription Factor, NFkappaB Biol Reprod, November 1, 2006; 75(5): 697 - 704. [Abstract] [Full Text] [PDF]

				M. S. Soloff, M. G. Izban, D. L. Cook Jr, Y.-J. Jeng, and R. C. Mifflin Interleukin-1-induced NF-{kappa}B recruitment to the oxytocin receptor gene inhibits RNA polymerase II-promoter interactions in cultured human myometrial cells Mol. Hum. Reprod., October 1, 2006; 12(10): 619 - 624. [Abstract] [Full Text] [PDF]

				M. D. Ashworth, J. W. Ross, J. Hu, F. J. White, D. R. Stein, U. DeSilva, G. A. Johnson, T. E. Spencer, and R. D. Geisert Expression of Porcine Endometrial Prostaglandin Synthase During the Estrous Cycle and Early Pregnancy, and Following Endocrine Disruption of Pregnancy Biol Reprod, June 1, 2006; 74(6): 1007 - 1015. [Abstract] [Full Text] [PDF]

				J. C. Condon, D. B. Hardy, K. Kovaric, and C. R. Mendelson Up-Regulation of the Progesterone Receptor (PR)-C Isoform in Laboring Myometrium by Activation of Nuclear Factor-{kappa}B May Contribute to the Onset of Labor through Inhibition of PR Function Mol. Endocrinol., April 1, 2006; 20(4): 764 - 775. [Abstract] [Full Text] [PDF]

				T. M Lindstrom and P. R Bennett The role of nuclear factor kappa B in human labour Reproduction, November 1, 2005; 130(5): 569 - 581. [Abstract] [Full Text] [PDF]

				N. R. Chapman, I. Smyrnias, D. O. C. Anumba, G. N. Europe-Finner, and S. C. Robson Expression of the GTP-Binding Protein (G{alpha}s) Is Repressed by the Nuclear Factor {kappa}B RelA Subunit in Human Myometrium Endocrinology, November 1, 2005; 146(11): 4994 - 5002. [Abstract] [Full Text] [PDF]

				Z. Strakova, P. Mavrogianis, X. Meng, J. M. Hastings, K. S. Jackson, P. Cameo, A. Brudney, O. Knight, and A. T. Fazleabas In Vivo Infusion of Interleukin-1{beta} and Chorionic Gonadotropin Induces Endometrial Changes that Mimic Early Pregnancy Events in the Baboon Endocrinology, September 1, 2005; 146(9): 4097 - 4104. [Abstract] [Full Text] [PDF]

				T. M Lindstrom and P. R Bennett 15-Deoxy-{Delta}12,14-Prostaglandin J2 Inhibits Interleukin-1{beta}-Induced Nuclear Factor-{kappa}B in Human Amnion and Myometrial Cells: Mechanisms and Implications J. Clin. Endocrinol. Metab., June 1, 2005; 90(6): 3534 - 3543. [Abstract] [Full Text] [PDF]

				N. R. Chapman, G. N. Europe-Finner, and S. C. Robson Expression and Deoxyribonucleic Acid-Binding Activity of the Nuclear Factor {kappa}B Family in the Human Myometrium during Pregnancy and Labor J. Clin. Endocrinol. Metab., November 1, 2004; 89(11): 5683 - 5693. [Abstract] [Full Text] [PDF]

				M. S. Soloff, Y.-J. Jeng, M. Ilies, S. L. Soloff, M. G. Izban, T. G. Wood, N. I. Panova, G. V.N. Velagaleti, and G. D. Anderson Immortalization and characterization of human myometrial cells from term-pregnant patients using a telomerase expression vector Mol. Hum. Reprod., September 1, 2004; 10(9): 685 - 695. [Abstract] [Full Text] [PDF]