DNA-binding ability of NF-kappaB is affected differently by ERalpha and ERbeta and its activation results in inhibition of estrogen responsiveness

Estrogenic effects involve interactions between estrogen receptors (ERs), response elements, and nuclear proteins. It is hypothesized that interaction between ER and NF-κB may affect the regulation of responsive genes. Electrophoretic mobility shift assay (EMSA) was performed to assess if the interaction of ERs and NF-κB affect their respective DNA-binding activities, and alkaline phosphatase assay was done to evaluate estrogenic activity. EMSA revealed that ERs inhibit DNA-binding of p50 and p65, whereas p50 did not impair ERα binding. Stimulation with estradiol inhibited DNA binding of NF-κB in ERα-transfected endometrial stromal cells (ESCs). Moreover, activation of NF-κB significantly decreased estrogen responsiveness of Ishikawa cells and ERα-transfected ESC. Our results suggest that ERs downregulate NF-κB-dependent gene activation by directly preventing DNA binding. However, NF-#x03BA;B-mediated inhibition of ER-dependent gene activation may be carried out indirectly rather than through a direct inhibition of ER-DNA binding. These findings offer new insight into the specific role of ERα and could eventually help in developing therapeutics for endometriosis. Similar content being viewed by others

Gruber CJ, Tschugguel W, Schneeberger C, Huber JC. Production and actions of estrogens. N Engl J Med. 2002; 346:340–352. DeMayo FJ, Zhao B, Takamoto N, Tsai SY. Mechanisms of action of estrogen and progesterone. Ann N Y Acad Sci. 2002; 955:48–59. Enmark E, Gustafsson JA. Oestrogen receptors-an overview. J Intern Med. 1999;246:133–138. Salama SA, Nasr AB, Dubey RK, Al-Hendy A. Estrogen metabolite 2-methoxyestradiol induces apoptosis and inhibits cell proliferation and collagen production in rat and human leiomyoma cells: a potential medicinal treatment for uterine fibroids. J Soc Gynecol Investig. 2006;13:542–550. Nilsson S, Gustafsson JA. Estrogen receptor action. Crit Rev Eukaryot Gene Expr. 2002;12:237–257. Sanchez R, Nguyen D, Rocha W, White JH, Mader S. Diversity in the mechanisms of gene regulation by estrogen receptors. Bioessays. 2002;24:244–254. Young SL. Complexity in endometrial estradiol signaling. Reprod Sci. 2007;14:627–628. Bharti AC, Aggarwal BB. Nuclear factor-kappa B and cancer: its role in prevention and therapy. Biochem Pharmacol. 2002;64: 883–888. Wang T, Zhang X, Li JJ. The role of NF-kappaB in the regulation of cell stress responses. Int Immunopharmacol. 2002;2: 1509–1520. Crossin KL. Cell adhesion molecules activate signaling networks that influence proliferation, gene expression, and differentiation. Ann N Y Acad Sci. 2002;961:159–160. Richmond A. NF-kappa B, chemokine gene transcription and tumour growth. Nat Rev Immunol. 2002;2:664–674. Oral E, Arici A. Pathogenesis of endometriosis. Obstet Gynecol Clin North Am. 1997;24:219–233. Ulukus M, Cakmak H, Arici A. The role of endometrium in endometriosis. J Soc Gynecol Investig. 2006;13:467–476. Halme J, Hammond MG, Hulka JF, Raj SG, Talbert LM. Retrograde menstruation in healthy women and in patients with endometriosis. Obstet Gynecol. 1984;64:151–154. Kruitwagen RF, Poels LG, Willemsen WN, Jap PH, Thomas CM, Rolland R. Retrograde seeding of endometrial epithelial cells by uterine-tubal flushing. Fertil Steril. 1991;56:414–420. Arici A, Senturk LM, Seli E, Bahtiyar MO, Kim G. Regulation of monocyte chemotactic protein-1 expression in human endometrial stromal cells by estrogen and progesterone. Biol Reprod. 1999;61:85–90. Kelly RW, King AE, Critchley HO. Cytokine control in human endometrium. Reproduction. 2001;121:3–19. Gielen SC, Santegoets LA, Kuhne LC, et al. Genomic and nongenomic effects of estrogen signaling in human endometrial cells: involvement of the growth factor receptor signaling downstream AKT pathway. Reprod Sci. 2007;14:646–654. Guzeloglu Kayisli O, Kayisli UA, Luleci G, Arici A. In vivo and in vitro regulation of Akt activation in human endometrial cells is estrogen dependent. Biol Reprod. 2004;71:714–721. Kalaitzidis D, Gilmore TD. Transcription factor cross-talk: the estrogen receptor and NF-kappaB. Trends Endocrinol Metab. 2005;16:46–52. Arici A, Head JR, MacDonald PC, Casey ML. Regulation of interleukin-8 gene expression in human endometrial cells in culture. Mol Cell Endocrinol. 1993;94:195–204. Littlefield BA, Gurpide E, Markiewicz L, McKinley B, Hochberg RB. A simple and sensitive microtiter plate estrogen bioassay based on stimulation of alkaline phosphatase in Ishikawa cells: estrogenic action of delta 5 adrenal steroids. Endocrinology. 1990;127:2757–2762. Lenardo MJ, Kuang A, Gifford A, Baltimore D. NF-kappa B protein purification from bovine spleen: nucleotide stimulation and binding site specificity. Proc Natl Acad Sci U S A. 1988;85:8825–8829. Osborn L, Kunkel S, Nabel GJ. Tumor necrosis factor alpha and interleukin 1 stimulate the human immunodeficiency virus enhancer by activation of the nuclear factor kappa B. Proc Natl Acad Sci U S A. 1989;86:2336–2340. Wierman ME, Kohrt WM. Vascular and metabolic effects of sex steroids: new insights into clinical trials. Reprod Sci. 2007;14:300–314. Singh RR, Kumar R. Steroid hormone receptor signaling in tumorigenesis. J Cell Biochem. 2005;96:490–505. Blobel GA, Sieff CA, Orkin SH. Ligand-dependent repression of the erythroid transcription factor GATA-1 by the estrogen receptor. Mol Cell Biol. 1995;15:3147–3153. Galien R, Evans HF, Garcia T. Involvement of CCAAT/ enhancer-binding protein and nuclear factor-kappa B binding sites in interleukin-6 promoter inhibition by estrogens. Mol Endocrinol. 1996;10:713–722. Turner RT, Riggs BL, Spelsberg TC. Skeletal effects of estrogen. Endocr Rev. 1994;15:275–300. McKay LI, Cidlowski JA. Molecular control of immune/ inflammatory responses: interactions between nuclear factor-kappa B and steroid receptor-signaling pathways. Endocr Rev. 1999;20:435–459. Caldenhoven E, Liden J, Wissink S, et al. Negative cross-talk between RelA and the glucocorticoid receptor: a possible mechanism for the anti-inflammatory action of glucocorticoids. Mol Endocrinol. 1995;9:401–412. Scheinman RI, Gualberto A, Jewell CM, Cidlowski JA, Baldwin AS Jr. Characterization of mechanisms involved in transrepression of NF-kappa B by activated glucocorticoid receptors. Mol Cell Biol. 1995;15:943–953. Ray P, Ghosh SK, Zhang DH, Ray A. Repression of interleukin-6 gene expression by 17 beta-estradiol: inhibition of the DNA-binding activity of the transcription factors NF-IL6 and NF-kappa B by the estrogen receptor. FEBS Lett. 1997;409:79–85. Boyce BF, Xing L, Franzoso G, Siebenlist U. Required and nonessential functions of nuclear factor-kappa B in bone cells. Bone. 1999;25:137–139. Stein B, Yang MX. Repression of the interleukin-6 promoter by estrogen receptor is mediated by NF-kappa B and C/EBP beta. Mol Cell Biol. 1995;15:4971–4979. Nakamura H, Kimura T, Ogita K, et al. Alteration of the timing of implantation by in vivo gene transfer: delay of implantation by suppression of nuclear factor kappaB activity and partial rescue by leukemia inhibitory factor. Biochem Biophys Res Commun. 2004;321:886–892. King AE, Critchley HO, Kelly RW. The NF-kappaB pathway in human endometrium and first trimester decidua. Mol Hum Reprod. 2001;7:175–183. Gonzalez-Ramos R, Donnez J, Defrere S, et al. Nuclear factor-kappa B is constitutively activated in peritoneal endometriosis. Mol Hum Reprod. 2007;13:503–509. Harris HA. Estrogen receptor-beta: recent lessons from in vivo studies. Mol Endocrinol. 2007;21:1–13. Khan KN, Masuzaki H, Fujishita A, et al. Interleukin-6- and tumour necrosis factor alpha-mediated expression of hepatocyte growth factor by stromal cells and its involvement in the growth of endometriosis. Hum Reprod. 2005;20: 2715–2723. Luk J, Seval Y, Kayisli UA, Ulukus M, Ulukus CE, Arici A. Regulation of interleukin-8 expression in human endometrial endothelial cells: a potential mechanism for the pathogenesis of endometriosis. J Clin Endocrinol Metab. 2005;90: 1805–1811. Sha G, Wu D, Zhang L, et al. Differentially expressed genes in human endometrial endothelial cells derived from eutopic endometrium of patients with endometriosis compared with those from patients without endometriosis. Hum Reprod. 2007; 22:3159–3169. Bukulmez O, Hardy DB, Carr BR, Word RA, Mendelson CR. Inflammatory status influences aromatase and steroid receptor expression in endometriosis. Endocrinology. 2008; 149:1190–1204. Jackson KS, Brudney A, Hastings JM, Mavrogianis PA, Kim JJ, Fazleabas AT. The altered distribution of the steroid hormone receptors and the chaperone immunophilin FKBP52 in a baboon model of endometriosis is associated with progesterone resistance during the window of uterine receptivity. Reprod Sci. 2007;14:137–150. Xue Q, Lin Z, Cheng YH, et al. Promoter methylation regulates estrogen receptor 2 in human endometrium and endometriosis. Biol Reprod. 2007;77:681–687. Sharma RV, Gurjar MV, Bhalla RC. Selected contribution: estrogen receptor-alpha gene transfer inhibits proliferation and NF-kappaB activation in VSM cells from female rats. J Appl Physiol. 2001;91:2400–2406. Speir E, Yu ZX, Takeda K, Ferrans VJ, Cannon RO, 3rd. Antioxidant effect of estrogen on cytomegalovirus-induced gene expression in coronary artery smooth muscle cells. Circulation. 2000;102:2990–2996. Hsu SM, Chen YC, Jiang MC. 17-Beta-estradiol inhibits tumor necrosis factor-alpha-induced nuclear factor-kappa B activation by increasing nuclear factor-kappa B p105 level in MCF-7 breast cancer cells. Biochem Biophys Res Commun. 2000;279:47–52. Lee YW, Eum SY, Nath A, Toborek M. Estrogen-mediated protection against HIV Tat protein-induced inflammatory pathways in human vascular endothelial cells. Cardiovasc Res. 2004;63:139–148. Dodel RC, Du Y, Bales KR, Gao F, Paul SM. Sodium salicylate and 17beta-estradiol attenuate nuclear transcription factor NF-kappaB translocation in cultured rat astroglial cultures following exposure to amyloid A beta(1-40) and lipopolysaccharides. J Neurochem. 1999;73:1453–1460. Author information Authors and Affiliations Corresponding author Additional information This study was supported by a training grant to GH (Ha-3375/1-1) from the Deutsche Forschungsgemeinschaft (DFG). Rights and permissions About this article Cite this article Guzeloglu-Kayisli, O., Halis, G., Taskiran, S. et al. DNA-Binding Ability of NF-κB is Affected Differently by ERα and ERβ and Its Activation Results in Inhibition of Estrogen Responsiveness. Reprod. Sci. 15, 493–505 (2008). https://doi.org/10.1177/1933719108317583 Published: Issue date: DOI: https://doi.org/10.1177/1933719108317583