Skip to main content
Key Specialties
Cardiology Diabetology Endocrinology Gastroenterology Geriatrics and Gerontology Gynecology and Obstetrics Hematology Infectious Disease Internal Medicine Nephrology Neurology Oncology Pediatrics Respiratory Medicine Rheumatology Surgery
Congress Coverage
2024 ACC Congress 2023 ESMO Congress 2023 EASD Congress 2023 ERS Congress 2023 ESC Congress
Clinical Collections
Advances in Alzheimer’s

At a glance: The ONWARDS insulin icodec trials

A round-up of the ONWARDS phase 3 clinical trials evaluating the novel weekly insulin icodec in people with diabetes.
ADVANCED SEARCH
Log in
Top
Breast Cancer Research
Published in: Breast Cancer Research 2/2013

Open Access 01-04-2013 | Research article

HIF-1α stimulates aromatase expression driven by prostaglandin E2 in breast adipose stroma

Authors: Nirukshi U Samarajeewa, Fangyuan Yang, Maria M Docanto, Minako Sakurai, Keely M McNamara, Hironobu Sasano, Stephen B Fox, Evan R Simpson, Kristy A Brown

Published in: Breast Cancer Research | Issue 2/2013

Login to get access

Abstract

Introduction

The majority of postmenopausal breast cancers are estrogen-dependent. Tumor-derived factors, such as prostaglandin E2 (PGE2), stimulate CREB1 binding to cAMP response elements (CREs) on aromatase promoter II (PII), leading to the increased expression of aromatase and biosynthesis of estrogens within human breast adipose stromal cells (ASCs). Hypoxia inducible factor-1α (HIF-1α), a key mediator of cellular adaptation to low oxygen levels, is emerging as a novel prognostic marker in breast cancer. We have identified the presence of a consensus HIF-1α binding motif overlapping with the proximal CRE of aromatase PII. However, the regulation of aromatase expression by HIF-1α in breast cancer has not been characterized. This study aimed to characterize the role of HIF-1α in the activation of aromatase PII.

Methods

HIF-1α expression and localization were examined in human breast ASCs using quantitative PCR (QPCR), Western blotting, immunofluorescence and high content screening. QPCR and tritiated water-release assays were performed to assess the effect of HIF-1α on aromatase expression and activity. Reporter assays and chromatin immunoprecipitation (ChIP) were performed to assess the effect of HIF-1α on PII activity and binding. Treatments included PGE2 or DMOG ((dimethyloxalglycine), HIF-1α stabilizer). Double immunohistochemistry for HIF-1α and aromatase was performed on tissues obtained from breast cancer and cancer-free patients.

Results

Results indicate that PGE2 increases HIF-1α transcript and protein expression, nuclear localization and binding to aromatase PII in human breast ASCs. Results also demonstrate that HIF-1α significantly increases PII activity, and aromatase transcript expression and activity, in the presence of DMOG and/or PGE2, and that HIF-1α and CREB1 act co-operatively on PII. There is a significant increase in HIF-1α positive ASCs in breast cancer patients compared to cancer-free women, and a positive association between HIF-1α and aromatase expression.

Conclusions

This study is the first to identify HIF-1α as a modulator of PII-driven aromatase expression in human breast tumor-associated stroma and provides a novel mechanism for estrogen regulation in obesity-related, post-menopausal breast cancer. Together with our on-going studies on the role of AMP-activated protein kinase (AMPK) in the regulation of breast aromatase, this work provides another link between disregulated metabolism and breast cancer.
Appendix
Available only for authorised users
Literature
1.
Stephenson GD, Rose DP: Breast cancer and obesity: an update. Nutr Cancer. 2003, 45: 1-16. 10.1207/S15327914NC4501_1.CrossRefPubMed
2.
Simpson ER, Brown KA: Obesity, aromatase and breast cancer. Expert Rev Endocrinol Metab. 2011, 6: 383-395. 10.1586/eem.11.35.CrossRef
3.
Agarwal VR, Bulun SE, Leitch M, Rohrich R, Simpson ER: Use of alternative promoters to express the aromatase cytochrome P450 (CYP19) gene in breast adipose tissues of cancer-free and breast cancer patients. J Clin Endocrinol Metab. 1996, 81: 3843-3849. 10.1210/jc.81.11.3843.PubMed
4.
Zhao Y, Agarwal VR, Mendelson CR, Simpson ER: Estrogen biosynthesis proximal to a breast tumor is stimulated by PGE2 via cyclic AMP, leading to activation of promoter II of the CYP19 (aromatase) gene. Endocrinology. 1996, 137: 5739-5742. 10.1210/en.137.12.5739.PubMed
5.
Subbaramaiah K, Morris PG, Zhou XK, Morrow M, Du B, Giri D, Kopelovich L, Hudis CA, Dannenberg AJ: Increased levels of COX-2 and prostaglandin E2 contribute to elevated aromatase expression in inflamed breast tissue of obese women. Cancer Discov. 2012, 2: 356-365. 10.1158/2159-8290.CD-11-0241.CrossRefPubMedPubMedCentral
6.
Sofi M, Young MJ, Papamakarios T, Simpson ER, Clyne CD: Role of CRE-binding protein (CREB) in aromatase expression in breast adipose. Breast Cancer Res Treat. 2003, 79: 399-407. 10.1023/A:1024038632570.CrossRefPubMed
7.
Brown KA, McInnes KJ, Hunger NI, Oakhill JS, Steinberg GR, Simpson ER: Subcellular localization of cyclic AMP-responsive element binding protein-regulated transcription coactivator 2 provides a link between obesity and breast cancer in postmenopausal women. Cancer Res. 2009, 69: 5392-5399. 10.1158/0008-5472.CAN-09-0108.CrossRefPubMed
8.
Zhou JF, Liu WL, Chen G, Lu YP, Ma D: [Molecular mechanism of inhibition of sodium butyrate on activation of tumor associated aromatase promoters]. Ai Zheng. 2002, 21: 1283-1287.PubMed
9.
Subbaramaiah K, Hudis C, Chang SH, Hla T, Dannenberg AJ: EP2 and EP4 receptors regulate aromatase expression in human adipocytes and breast cancer cells. Evidence of a BRCA1 and p300 exchange. J Biol Chem. 2008, 283: 3433-3444.CrossRefPubMed
10.
Vaupel P, Briest S, Hockel M: Hypoxia in breast cancer: pathogenesis, characterization and biological/therapeutic implications. Wien Med Wochenschr. 2002, 152: 334-342. 10.1046/j.1563-258X.2002.02032.x.CrossRefPubMed
11.
Semenza GL: Hypoxia, clonal selection, and the role of HIF-1 in tumor progression. Crit Rev Biochem Mol Biol. 2000, 35: 71-103. 10.1080/10409230091169186.CrossRefPubMed
12.
Generali D, Buffa FM, Berruti A, Brizzi MP, Campo L, Bonardi S, Bersiga A, Allevi G, Milani M, Aguggini S, Papotti M, Dogliotti L, Bottini A, Harris AL, Fox SB: Phosphorylated ERalpha, HIF-1alpha, and MAPK signaling as predictors of primary endocrine treatment response and resistance in patients with breast cancer. J Clin Oncol. 2009, 27: 227-234. 10.1200/JCO.2007.13.7083.CrossRefPubMed
13.
Yamamoto Y, Ibusuki M, Okumura Y, Kawasoe T, Kai K, Iyama K, Iwase H: Hypoxia-inducible factor 1alpha is closely linked to an aggressive phenotype in breast cancer. Breast Cancer Res Treat. 2008, 110: 465-475. 10.1007/s10549-007-9742-1.CrossRefPubMed
14.
Melillo G: Targeting hypoxia cell signaling for cancer therapy. Cancer Metastasis Rev. 2007, 26: 341-352. 10.1007/s10555-007-9059-x.CrossRefPubMed
15.
16.
Wang GL, Jiang BH, Rue EA, Semenza GL: Hypoxia-inducible factor 1 is a basic-helix-loop-helix-PAS heterodimer regulated by cellular O2 tension. Proc Natl Acad Sci USA. 1995, 92: 5510-5514. 10.1073/pnas.92.12.5510.CrossRefPubMedPubMedCentral
17.
Salceda S, Caro J: Hypoxia-inducible factor 1alpha (HIF-1alpha) protein is rapidly degraded by the ubiquitin-proteasome system under normoxic conditions. Its stabilization by hypoxia depends on redox-induced changes. J Biol Chem. 1997, 272: 22642-22647. 10.1074/jbc.272.36.22642.CrossRefPubMed
18.
Huang LE, Gu J, Schau M, Bunn HF: Regulation of hypoxia-inducible factor 1alpha is mediated by an O2-dependent degradation domain via the ubiquitin-proteasome pathway. Proc Natl Acad Sci USA. 1998, 95: 7987-7992. 10.1073/pnas.95.14.7987.CrossRefPubMedPubMedCentral
19.
Semenza GL, Jiang BH, Leung SW, Passantino R, Concordet JP, Maire P, Giallongo A: Hypoxia response elements in the aldolase A, enolase 1, and lactate dehydrogenase A gene promoters contain essential binding sites for hypoxia-inducible factor 1. J Biol Chem. 1996, 271: 32529-32537. 10.1074/jbc.271.51.32529.CrossRefPubMed
20.
Arany Z, Huang LE, Eckner R, Bhattacharya S, Jiang C, Goldberg MA, Bunn HF, Livingston DM: An essential role for p300/CBP in the cellular response to hypoxia. Proc Natl Acad Sci USA. 1996, 93: 12969-12973. 10.1073/pnas.93.23.12969.CrossRefPubMedPubMedCentral
21.
Harris AL: Hypoxia--a key regulatory factor in tumour growth. Nat Rev Cancer. 2002, 2: 38-47. 10.1038/nrc704.CrossRefPubMed
22.
Liu XH, Kirschenbaum A, Lu M, Yao S, Dosoretz A, Holland JF, Levine AC: Prostaglandin E2 induces hypoxia-inducible factor-1alpha stabilization and nuclear localization in a human prostate cancer cell line. J Biol Chem. 2002, 277: 50081-50086. 10.1074/jbc.M201095200.CrossRefPubMed
23.
Fukuda R, Kelly B, Semenza GL: Vascular endothelial growth factor gene expression in colon cancer cells exposed to prostaglandin E2 is mediated by hypoxia-inducible factor 1. Cancer Res. 2003, 63: 2330-2334.PubMed
24.
Michael MD, Kilgore MW, Morohashi K, Simpson ER: Ad4BP/SF-1 regulates cyclic AMP-induced transcription from the proximal promoter (PII) of the human aromatase P450 (CYP19) gene in the ovary. J Biol Chem. 1995, 270: 13561-13566. 10.1074/jbc.270.22.13561.CrossRefPubMed
25.
Kondo K, Klco J, Nakamura E, Lechpammer M, Kaelin WG: Inhibition of HIF is necessary for tumor suppression by the von Hippel-Lindau protein. Cancer Cell. 2002, 1: 237-246. 10.1016/S1535-6108(02)00043-0.CrossRefPubMed
26.
Ackerman GE, Smith ME, Mendelson CR, MacDonald PC, Simpson ER: Aromatization of androstenedione by human adipose tissue stromal cells in monolayer culture. J Clin Endocrinol Metab. 1981, 53: 412-417. 10.1210/jcem-53-2-412.CrossRefPubMed
27.
Brown KA, Hunger NI, Docanto M, Simpson ER: Metformin inhibits aromatase expression in human breast adipose stromal cells via stimulation of AMP-activated protein kinase. Breast Cancer Res Treat. 2010, 123: 591-596. 10.1007/s10549-010-0834-y.CrossRefPubMed
28.
29.
Hong Y, Li H, Ye J, Miki Y, Yuan YC, Sasano H, Evans DB, Chen S: Epitope characterization of an aromatase monoclonal antibody suitable for the assessment of intratumoral aromatase activity. PLoS One. 2009, 4: e8050-10.1371/journal.pone.0008050.CrossRefPubMedPubMedCentral
30.
Messeguer X, Escudero R, Farre D, Nunez O, Martinez J, Alba MM: PROMO: detection of known transcription regulatory elements using species-tailored searches. Bioinformatics. 2002, 18: 333-334. 10.1093/bioinformatics/18.2.333.CrossRefPubMed
31.
Farré D, Roset R, Huerta M, Adsuara JE, Roselló L, Albà MM, Messeguer X: Identification of patterns in biological sequences at the ALGGEN server: PROMO and MALGEN. Nucleic Acids Res. 2003, 31: 3651-3653. 10.1093/nar/gkg605.CrossRefPubMedPubMedCentral
32.
Messeguer X, Escudero R, Farré D, Nuñez O, Martínez J, Albà MM: PROMO: detection of known transcription regulatory elements using species-tailored searches. Bioinformatics. 2002, 18: 333-334. 10.1093/bioinformatics/18.2.333.CrossRefPubMed
33.
Zhong H, De Marzo AM, Laughner E, Lim M, Hilton DA, Zagzag D, Buechler P, Isaacs WB, Semenza GL, Simons JW: Overexpression of hypoxia-inducible factor 1alpha in common human cancers and their metastases. Cancer Res. 1999, 59: 5830-5835.PubMed
34.
35.
Subbaramaiah K, Howe LR, Port ER, Brogi E, Fishman J, Liu CH, Hla T, Hudis C, Dannenberg AJ: HER-2/neu status is a determinant of mammary aromatase activity in vivo: evidence for a cyclooxygenase-2-dependent mechanism. Cancer Res. 2006, 66: 5504-5511. 10.1158/0008-5472.CAN-05-4076.CrossRefPubMed
36.
Huang SP, Wu MS, Shun CT, Wang HP, Hsieh CY, Kuo ML, Lin JT: Cyclooxygenase-2 increases hypoxia-inducible factor-1 and vascular endothelial growth factor to promote angiogenesis in gastric carcinoma. J Biomed Sci. 2005, 12: 229-241. 10.1007/s11373-004-8177-5.CrossRefPubMed
37.
Ji R, Chou CL, Xu W, Chen XB, Woodward DF, Regan JW: EP1 prostanoid receptor coupling to G i/o up-regulates the expression of hypoxia-inducible factor-1 alpha through activation of a phosphoinositide-3 kinase signaling pathway. Mol Pharmacol. 2010, 77: 1025-1036. 10.1124/mol.110.063933.CrossRefPubMedPubMedCentral
38.
Richards JA, Brueggemeier RW: Prostaglandin E2 regulates aromatase activity and expression in human adipose stromal cells via two distinct receptor subtypes. J Clin Endocrinol Metab. 2003, 88: 2810-2816. 10.1210/jc.2002-021475.CrossRefPubMed
39.
Kumar P, Mendelson CR: Estrogen-related receptor gamma (ERRgamma) mediates oxygen-dependent induction of aromatase (CYP19) gene expression during human trophoblast differentiation. Mol Endocrinol. 2011, 25: 1513-1526. 10.1210/me.2011-1012.CrossRefPubMedPubMedCentral
40.
Dimova EY, Jakubowska MM, Kietzmann T: CREB binding to the hypoxia-inducible factor-1 responsive elements in the plasminogen activator inhibitor-1 promoter mediates the glucagon effect. Thromb Haemost. 2007, 98: 296-303.PubMed
41.
Choi JH, Cho HK, Choi YH, Cheong J: Activating transcription factor 2 increases transactivation and protein stability of hypoxia-inducible factor 1alpha in hepatocytes. Biochem J. 2009, 424: 285-296. 10.1042/BJ20090371.CrossRefPubMed
42.
Choi JH, Park MJ, Kim KW, Choi YH, Park SH, An WG, Yang US, Cheong J: Molecular mechanism of hypoxia-mediated hepatic gluconeogenesis by transcriptional regulation. FEBS Lett. 2005, 579: 2795-2801. 10.1016/j.febslet.2005.03.097.CrossRefPubMed
43.
Freedman SJ, Sun ZY, Poy F, Kung AL, Livingston DM, Wagner G, Eck MJ: Structural basis for recruitment of CBP/p300 by hypoxia-inducible factor-1 alpha. Proc Natl Acad Sci USA. 2002, 99: 5367-5372. 10.1073/pnas.082117899.CrossRefPubMedPubMedCentral
44.
Chhabra A, Fernando H, Watkins G, Mansel RE, Jiang WG: Expression of transcription factor CREB1 in human breast cancer and its correlation with prognosis. Oncol Rep. 2007, 18: 953-958.PubMed
45.
Sofi M, Young MJ, Papamakarios T, Simpson ER, Clyne CD: Role of CRE-binding protein (CREB) in aromatase expression in breast adipose. Breast Cancer Res Treat. 2003, 79: 399-407. 10.1023/A:1024038632570.CrossRefPubMed
46.
Bonneterre J, Buzdar A, Nabholtz JM, Robertson JF, Thurlimann B, von Euler M, Sahmoud T, Webster A, Steinberg M: Anastrozole is superior to tamoxifen as first-line therapy in hormone receptor positive advanced breast carcinoma. Cancer. 2001, 92: 2247-2258. 10.1002/1097-0142(20011101)92:9<2247::AID-CNCR1570>3.0.CO;2-Y.CrossRefPubMed
47.
Mouridsen H, Gershanovich M, Sun Y, Perez-Carrion R, Boni C, Monnier A, Apffelstaedt J, Smith R, Sleeboom HP, Janicke F, Pluzanska A, Dank M, Becquart D, Bapsy PP, Salminen E, Snyder R, Lassus M, Verbeek JA, Staffler B, Chaudri-Ross HA, Dugan M: Superior efficacy of letrozole versus tamoxifen as first-line therapy for postmenopausal women with advanced breast cancer: results of a phase III study of the International Letrozole Breast Cancer Group. J Clin Oncol. 2001, 19: 2596-2606.PubMed
48.
Nabholtz JM, Buzdar A, Pollak M, Harwin W, Burton G, Mangalik A, Steinberg M, Webster A, von Euler M: Anastrozole is superior to tamoxifen as first-line therapy for advanced breast cancer in postmenopausal women: results of a North American multicenter randomized trial. Arimidex Study Group. J Clin Oncol. 2000, 18: 3758-3767.PubMed
49.
Mouridsen HT: Incidence and management of side effects associated with aromatase inhibitors in the adjuvant treatment of breast cancer in postmenopausal women. Curr Med Res Opin. 2006, 22: 1609-1621. 10.1185/030079906X115667.CrossRefPubMed
50.
Henry NL, Azzouz F, Desta Z, Li L, Nguyen AT, Lemler S, Hayden J, Tarpinian K, Yakim E, Flockhart DA, Stearns V, Hayes DF, Storniolo AM: Predictors of aromatase inhibitor discontinuation as a result of treatment-emergent symptoms in early-stage breast cancer. J Clin Oncol. 2012, 30: 936-942. 10.1200/JCO.2011.38.0261.CrossRefPubMedPubMedCentral
51.
Henry NL, Giles JT, Ang D, Mohan M, Dadabhoy D, Robarge J, Hayden J, Lemler S, Shahverdi K, Powers P, Li L, Flockhart D, Stearns V, Hayes DF, Storniolo AM, Clauw DJ: Prospective characterization of musculoskeletal symptoms in early stage breast cancer patients treated with aromatase inhibitors. Breast Cancer Res Treat. 2008, 111: 365-372. 10.1007/s10549-007-9774-6.CrossRefPubMed
52.
Onnis B, Rapisarda A, Melillo G: Development of HIF-1 inhibitors for cancer therapy. J Cell Mol Med. 2009, 13: 2780-2786. 10.1111/j.1582-4934.2009.00876.x.CrossRefPubMedPubMedCentral
Metadata
Title
HIF-1α stimulates aromatase expression driven by prostaglandin E2 in breast adipose stroma
Authors
Nirukshi U Samarajeewa
Fangyuan Yang
Maria M Docanto
Minako Sakurai
Keely M McNamara
Hironobu Sasano
Stephen B Fox
Evan R Simpson
Kristy A Brown
Publication date
01-04-2013
Publisher
BioMed Central
Published in
Breast Cancer Research / Issue 2/2013
Electronic ISSN: 1465-542X
DOI
https://doi.org/10.1186/bcr3410

Other articles of this Issue 2/2013

Breast Cancer Research 2/2013 Go to the issue

Research article

Pharmacologic reversion of epigenetic silencing of the PRKD1promoter blocks breast tumor cell invasion and metastasis

Viewpoint

Closing escape routes: inhibition of IL-8 signaling enhances the anti-tumor efficacy of PI3K inhibitors

Research article

Arthritis augments breast cancer metastasis: role of mast cells and SCF/c-Kit signaling

Editorial

It takes two to tango in the microenvironment!

Research article

Parity induces differentiation and reduces Wnt/Notch signaling ratio and proliferation potential of basal stem/progenitor cells isolated from mouse mammary epithelium

Research article

Oestrogen increases the activity of oestrogen receptor negative breast cancer stem cells through paracrine EGFR and Notch signalling
Webinar | 19-02-2024 | 17:30 (CET)

Keynote webinar | Spotlight on antibody–drug conjugates in cancer

Watch now

Antibody–drug conjugates (ADCs) are novel agents that have shown promise across multiple tumor types. Explore the current landscape of ADCs in breast and lung cancer with our experts, and gain insights into the mechanism of action, key clinical trials data, existing challenges, and future directions.

Dr. Véronique Diéras
Prof. Fabrice Barlesi
Developed by: Springer Medicine
Image Credits