IGF-I stimulates ERβ and aromatase expression via IGF1R/PI3K/AKT-mediated transcriptional activation in endometriosis

Estrogen receptor beta (ERβ, encoded by ESR2 gene) and cytochrome P450 aromatase (encoded by CYP19A1 gene) play critical roles in endometriosis, and the levels of insulin-like growth factor-I (IGF-I) in the peritoneal fluid are significantly higher in patients with endometriosis compared with those in normal women. However, the effects and mechanisms of IGF-I on ERβ and aromatase expression remain to be fully elucidated. In this study, human endometriotic stromal cells (ESCs) and endometrial cells (EMs) derived from ovarian endometriomas and eutopic endometrial tissues. ESCs were cultured with IGF-I, signal pathway inhibitors, and siRNAs. ERβ and aromatase expression were measured by real-time PCR and Western, respectively. The binding of c-Jun and CREB to the ESR2 and CYP19A1 promoters was assessed by chromatin immunoprecipitation assay. Animal experiments were performed in a xenograft mouse model. Levels of IGF-I mRNA in ESCs were markedly higher than those in EMs. IGF-I upregulated ERβ and aromatase expression in ESCs after stimulation of the IGF1R/PI3K/AKT pathway. Following IGF-I treatment, a marked increase in c-Jun and CREB phosphorylation occurred, enhancing binding to the ESR2 and CYP19A1 promoters. An IGF1R inhibitor in vivo reduced IGF-I-induced endometriosis graft growth and ERβ and aromatase expression. In conclusion, this is the first report to describe a mechanistic analysis of ERβ and aromatase expression regulated by IGF-I in ESCs. Moreover, an IGF1R inhibitor impeded ectopic lesion growth in nude mice. These findings suggest that an inhibitor of IGF1R might have therapeutic potential as an antiendometriotic drug. Key messages - Level of IGF-I mRNA in ESCs is markedly higher than that in EMs. - IGF-I up-regulates ERβ and aromatase expression via IGF1R/PI3K/AKT pathway. - C-Jun and CREB are recruited to ESR2 or CYP19A1 promoter by IGF-I stimulation. - IGF-1R inhibitors in vivo impede the growth of ectopic lesions in nude mice. Access this article We’re sorry, something doesn't seem to be working properly. Please try refreshing the page. If that doesn't work, please contact support so we can address the problem. Similar content being viewed by others

Olive DL, Schwartz LB (1993) Endometriosis. N Engl J Med 328:1759–1769 Ryan IP, Taylor RN (1997) Endometriosis and infertility: new concepts. Obstet Gynecol Surv 52:365–371 Zeitoun K, Takayama K, Michael MD, Bulun SE (1999) Stimulation of aromatase P450 promoter (II) activity in endometriosis and its inhibition in endometrium are regulated by competitive binding of steroidogenic factor-1 and chicken ovalbumin upstream promoter transcription factor to the same cis-acting element. Mol Endocrinol 13:239–253 Bulun SE, Yang S, Fang Z, Gurates B, Tamura M, Zhou J, Sebastian S (2001) Role of aromatase in endometrial disease. J Steroid Biochem Mol Biol 79:19–25 Xu JN, Zeng C, Zhou Y, Peng C, Zhou YF, Xue Q (2014) Metformin inhibits StAR expression in human endometriotic stromal cells via AMPK-mediated disruption of CREB-CRTC2 complex formation. J Clin Endocrinol Metab 99:2795–2803 Almeida M, Iyer S, Martin-Millan M, Bartell SM, Han L, Ambrogini E, Onal M, Xiong J, Weinstein RS, Jilka RL et al (2013) Estrogen receptor-alpha signaling in osteoblast progenitors stimulates cortical bone accrual. J Clin Invest 123:394–404 Hewitt SC, Harrell JC, Korach KS (2005) Lessons in estrogen biology from knockout and transgenic animals. Annu Rev Physiol 67:285–308 Trukhacheva E, Lin Z, Reierstad S, Cheng YH, Milad M, Bulun SE (2009) Estrogen receptor (ER) beta regulates ERalpha expression in stromal cells derived from ovarian endometriosis. J Clin Endocrinol Metab 94:615–622 Bulun SE, Cheng YH, Pavone ME, Xue Q, Attar E, Trukhacheva E, Tokunaga H, Utsunomiya H, Yin P, Luo X et al (2010) Estrogen receptor-beta, estrogen receptor-alpha, and progesterone resistance in endometriosis. Semin Reprod Med 28:36–43 Sebastian S, Bulun SE (2001) A highly complex organization of the regulatory region of the human CYP19 (aromatase) gene revealed by the Human Genome Project. J Clin Endocrinol Metab 86:4600–4602 Attar E, Tokunaga H, Imir G, Yilmaz MB, Redwine D, Putman M, Gurates B, Attar R, Yaegashi N, Hales DB et al (2009) Prostaglandin E2 via steroidogenic factor-1 coordinately regulates transcription of steroidogenic genes necessary for estrogen synthesis in endometriosis. J Clin Endocrinol Metab 94:623–631 Attar E, Bulun SE (2006) Aromatase and other steroidogenic genes in endometriosis: translational aspects. Hum Reprod Update 12:49–56 Li LC, Yeh CC, Nojima D, Dahiya R (2000) Cloning and characterization of human estrogen receptor beta promoter. Biochem Biophys Res Commun 275:682–689 Hinshelwood MM, Michael MD, Simpson ER (1997) The 5'-flanking region of the ovarian promoter of the bovine CYP19 gene contains a deletion in a cyclic adenosine 3',5'-monophosphate-like responsive sequence. Endocrinology 138:3704–3710 Michael MD, Michael LF, Simpson ER (1997) A CRE-like sequence that binds CREB and contributes to cAMP-dependent regulation of the proximal promoter of the human aromatase P450 (CYP19) gene. Mol Cell Endocrinol 134:147–156 Giudice LC, Dsupin BA, Gargosky SE, Rosenfeld RG, Irwin JC (1994) The insulin-like growth factor system in human peritoneal fluid: its effects on endometrial stromal cells and its potential relevance to endometriosis. J Clin Endocrinol Metab 79:1284–1293 Koutsilieris M, Mastrogamvrakis G, Lembessis P, Sourla A, Miligos S, Michalas S (2001) Increased insulin-like growth factor 1 activity can rescue KLE endometrial-like cells from apoptosis. Mol Med 7:20–26 Mu F, Hankinson SE, Schernhammer E, Pollak MN, Missmer SA (2015) A prospective study of insulin-like growth factor 1, its binding protein 3, and risk of endometriosis. Am J Epidemiol 182:148–156 Kim JG, Suh CS, Kim SH, Choi YM, Moon SY, Lee JY (2000) Insulin-like growth factors (IGFs), IGF-binding proteins (IGFBPs), and IGFBP-3 protease activity in the peritoneal fluid of patients with and without endometriosis. Fertil Steril 73:996–1000 LeRoith D, Roberts CT Jr (1993) Insulin-like growth factors. Ann N Y Acad Sci 692:1–9 Manna PR, Chandrala SP, King SR, Jo Y, Counis R, Huhtaniemi IT, Stocco DM (2006) Molecular mechanisms of insulin-like growth factor-I mediated regulation of the steroidogenic acute regulatory protein in mouse Leydig cells. Mol Endocrinol 20:362–378 Zuloaga R, Fuentes EN, Molina A, Valdes JA (2013) The cAMP response element binding protein (CREB) is activated by insulin-like growth factor-1 (IGF-1) and regulates myostatin gene expression in skeletal myoblast. Biochem Biophys Res Commun 440:258–264 Velarde MC, Aghajanova L, Nezhat CR, Giudice LC (2009) Increased mitogen-activated protein kinase kinase/extracellularly regulated kinase activity in human endometrial stromal fibroblasts of women with endometriosis reduces 3',5'-cyclic adenosine 5'-monophosphate inhibition of cyclin D1. Endocrinology 150:4701–4712 Yin X, Pavone ME, Lu Z, Wei J, Kim JJ (2012) Increased activation of the PI3K/AKT pathway compromises decidualization of stromal cells from endometriosis. J Clin Endocrinol Metab 97:E35–43 Reverchon M, Cornuau M, Rame C, Guerif F, Royere D, Dupont J (2013) Resistin decreases insulin-like growth factor I-induced steroid production and insulin-like growth factor I receptor signaling in human granulosa cells. Fertil Steril 100(247–255):e241–243 Rizza P, Barone I, Zito D, Giordano F, Lanzino M, De Amicis F, Mauro L, Sisci D, Catalano S, Dahlman Wright K et al (2014) Estrogen receptor beta as a novel target of androgen receptor action in breast cancer cell lines. Breast Cancer Res 16:R21 Reverchon M, Cornuau M, Rame C, Guerif F, Royere D, Dupont J (2012) Chemerin inhibits IGF-1-induced progesterone and estradiol secretion in human granulosa cells. Hum Reprod 27:1790–1800 Ryan IP, Schriock ED, Taylor RN (1994) Isolation, characterization, and comparison of human endometrial and endometriosis cells in vitro. J Clin Endocrinol Metab 78:642–649 Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(−Delta Delta C(T)) Method. Methods 25:402–408 Deb S, Zhou J, Amin SA, Imir AG, Yilmaz MB, Lin Z, Bulun SE (2006) A novel role of sodium butyrate in the regulation of cancer-associated aromatase promoters I.3 and II by disrupting a transcriptional complex in breast adipose fibroblasts. J Biol Chem 281:2585–2597 Bruner KL, Matrisian LM, Rodgers WH, Gorstein F, Osteen KG (1997) Suppression of matrix metalloproteinases inhibits establishment of ectopic lesions by human endometrium in nude mice. J Clin Invest 99:2851–2857 Bruner-Tran KL, Eisenberg E, Yeaman GR, Anderson TA, McBean J, Osteen KG (2002) Steroid and cytokine regulation of matrix metalloproteinase expression in endometriosis and the establishment of experimental endometriosis in nude mice. J Clin Endocrinol Metab 87:4782–4791 Eaton JL, Unno K, Caraveo M, Lu Z, Kim JJ (2013) Increased AKT or MEK1/2 activity influences progesterone receptor levels and localization in endometriosis. J Clin Endocrinol Metab 98:E1871–1879 Stewart CE, Rotwein P (1996) Growth, differentiation, and survival: multiple physiological functions for insulin-like growth factors. Physiol Rev 76:1005–1026 Hawsawi Y, El-Gendy R, Twelves C, Speirs V, Beattie J (2013) Insulin-like growth factor—oestradiol crosstalk and mammary gland tumourigenesis. Biochim Biophys Acta 1836:345–353 Milingos D, Katopodis H, Milingos S, Protopapas A, Creatsas G, Michalas S, Antsaklis A, Koutsilieris M (2006) Insulin-like growth factor-1 isoform mRNA expression in women with endometriosis: eutopic endometrium versus endometriotic cyst. Ann N Y Acad Sci 1092:434–439 Milingos DS, Philippou A, Armakolas A, Papageorgiou E, Sourla A, Protopapas A, Liapi A, Antsaklis A, Mastrominas M, Koutsilieris M (2011) Insulinlike growth factor-1Ec (MGF) expression in eutopic and ectopic endometrium: characterization of the MGF E-peptide actions in vitro. Mol Med 17:21–28 Tsonis AI, Afratis N, Gialeli C, Ellina MI, Piperigkou Z, Skandalis SS, Theocharis AD, Tzanakakis GN, Karamanos NK (2013) Evaluation of the coordinated actions of estrogen receptors with epidermal growth factor receptor and insulin-like growth factor receptor in the expression of cell surface heparan sulfate proteoglycans and cell motility in breast cancer cells. FEBS J 280:2248–2259 Mendoza RA, Enriquez MI, Mejia SM, Moody EE, Thordarson G (2011) Interactions between IGF-I, estrogen receptor-alpha (ERalpha), and ERbeta in regulating growth/apoptosis of MCF-7 human breast cancer cells. J Endocrinol 208:1–9 Sirianni R, Capparelli C, Chimento A, Panza S, Catalano S, Lanzino M, Pezzi V, Ando S (2012) Nandrolone and stanozolol upregulate aromatase expression and further increase IGF-I-dependent effects on MCF-7 breast cancer cell proliferation. Mol Cell Endocrinol 363:100–110 Rice S, Pellatt L, Ramanathan K, Whitehead SA, Mason HD (2009) Metformin inhibits aromatase via an extracellular signal-regulated kinase-mediated pathway. Endocrinology 150:4794–4801 Sirianni R, Chimento A, Malivindi R, Mazzitelli I, Ando S, Pezzi V (2007) Insulin-like growth factor-I, regulating aromatase expression through steroidogenic factor 1, supports estrogen-dependent tumor Leydig cell proliferation. Cancer Res 67:8368–8377 LeRoith D, Werner H, Beitner-Johnson D, Roberts CT Jr (1995) Molecular and cellular aspects of the insulin-like growth factor I receptor. Endocr Rev 16:143–163 Shazand K, Baban S, Prive C, Malette B, Croteau P, Lagace M, Racine JB, Hugo P (2004) FOXO1 and c-jun transcription factors mRNA are modulated in endometriosis. Mol Hum Reprod 10:871–877 Hull ML, Escareno CR, Godsland JM, Doig JR, Johnson CM, Phillips SC, Smith SK, Tavare S, Print CG, Charnock-Jones DS (2008) Endometrial-peritoneal interactions during endometriotic lesion establishment. Am J Pathol 173:700–715 Ohlsson Teague EM, Van der Hoek KH, Van der Hoek MB, Perry N, Wagaarachchi P, Robertson SA, Print CG, Hull LM (2009) MicroRNA-regulated pathways associated with endometriosis. Mol Endocrinol 23:265–275 Yang Y, Yee D (2012) Targeting insulin and insulin-like growth factor signaling in breast cancer. J Mammary Gland Biol Neoplasia 17:251–261 Tang H, Liao Y, Xu L, Zhang C, Liu Z, Deng Y, Jiang Z, Fu S, Chen Z, Zhou S (2013) Estrogen and insulin-like growth factor 1 synergistically promote the development of lung adenocarcinoma in mice. Int J Cancer 133:2473–2482 Yang Y, Yee D (2014) IGF-I regulates redox status in breast cancer cells by activating the amino acid transport molecule xC. Cancer Res 74:2295–2305 Zhou WD, Yang HM, Wang Q, Su DY, Liu FA, Zhao M, Chen QH, Chen QX (2010) SB203580, a p38 mitogen-activated protein kinase inhibitor, suppresses the development of endometriosis by down-regulating proinflammatory cytokines and proteolytic factors in a mouse model. Hum Reprod 25:3110–3116 Acknowledgments We appreciate Prof. Yu Qi and Prof. Ding-Fang Bu for their generous advices to the study. This work was supported by grants from the National Natural Science Foundation of China (Grant No. 81270674) and the Natural Science Foundation of Beijing, China (Grant No.7132204). Author information Authors and Affiliations Corresponding author Ethics declarations The cell experimental procedures were approved by the institutional review board of the First Hospital of Peking University (No. 2014[789] and No. 2014[790]), and signed informed consents for use of the samples were obtained from each patient. The First Hospital of Peking University Animal Care Committee approved the use of mice for this study (No. J201403). Conflict of interest The authors declare that they have no competing interests. Rights and permissions About this article Cite this article Zhou, Y., Zeng, C., Li, X. et al. IGF-I stimulates ERβ and aromatase expression via IGF1R/PI3K/AKT-mediated transcriptional activation in endometriosis. J Mol Med 94, 887–897 (2016). https://doi.org/10.1007/s00109-016-1396-1 Received: Revised: Accepted: Published: Issue date: DOI: https://doi.org/10.1007/s00109-016-1396-1