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BMC Cancer
Published in: BMC Cancer 1/2013

Open Access 01-12-2013 | Research article

Clinical instability of breast cancer markers is reflected in long-term in vitro estrogen deprivation studies

Authors: Jelena Milosevic, Johanna Klinge, Anna-Lena Borg, Theodoros Foukakis, Jonas Bergh, Nicholas P Tobin

Published in: BMC Cancer | Issue 1/2013

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Abstract

Background

Long-term estrogen deprivation models are widely employed in an in vitro setting to recapitulate the hormonal milieu of breast cancer patients treated with endocrine therapy. Despite the wealth information we have garnered from these models thus far, a comprehensive time-course analysis of the estrogen (ER), progesterone (PR), and human epidermal growth factor 2 (HER-2/neu) receptors on the gene and protein level, coupled with expression array data is currently lacking. We aimed to address this knowledge gap in order to enhance our understanding of endocrine therapy resistance in breast cancer patients.

Methods

ER positive MCF7 and BT474 breast cancer cells were grown in estrogen depleted medium for 10 months with the ER negative MDA-MB-231 cell line employed as control. ER, PR and HER-2/neu expression were analysed at defined short and long-term time points by immunocytochemistry (ICC), and quantitative real-time RT-PCR (qRT-PCR). Microarray analysis was performed on representative samples.

Results

MCF7 cells cultured in estrogen depleted medium displayed decreasing expression of ER up to 8 weeks, which was then re-expressed at 10 months. PR was also down-regulated at early time points and remained so for the duration of the study. BT474 cells generally displayed no changes in ER during the first 8 weeks of deprivation, however its expression was significantly decreased at 10 months. PR expression was also down-regulated early in BT474 samples and was absent at later time points. Finally, microarray data revealed that genes and cell processes down-regulated in both cell lines at 6 weeks overlapped with those down-regulated in aromatase inhibitor treated breast cancer patients.

Conclusions

Our data demonstrate that expression of ER, PR, and cell metabolic/proliferative processes are unstable in response to long-term estrogen deprivation in breast cancer cell lines. These results mirror recent clinical findings and again emphasize the utility of LTED models in translational research.
Appendix
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Literature
1.
Jordan VC: The ups and downs of the estrogen receptor. J Clin Oncol. 2003, 21: 3-4. 10.1200/JCO.2003.10.008.CrossRefPubMed
2.
Song RX, Mor G, Naftolin F, McPherson RA, Song J, Zhang Z, Yue W, Wang J, Santen RJ: Effect of long-term estrogen deprivation on apoptotic responses of breast cancer cells to 17beta-estradiol. J Natl Cancer Inst. 2001, 93: 1714-1723. 10.1093/jnci/93.22.1714.CrossRefPubMed
3.
Lippert C, Seeger H, Mueck AO: The effect of endogenous estradiol metabolites on the proliferation of human breast cancer cells. Life Sci. 2003, 72: 877-883. 10.1016/S0024-3205(02)02305-6.CrossRefPubMed
4.
Osborne CK, Schiff R: Estrogen-receptor biology: continuing progress and therapeutic implications. J Clin Oncol. 2005, 23: 1616-1622. 10.1200/JCO.2005.10.036.CrossRefPubMed
5.
Osborne CK: Steroid hormone receptors in breast cancer management. Breast Cancer Res Treat. 1998, 51: 227-238. 10.1023/A:1006132427948.CrossRefPubMed
6.
Kumar V, Green S, Stack G, Berry M, Jin JR, Chambon P: Functional domains of the human estrogen receptor. Cell. 1987, 51: 941-951. 10.1016/0092-8674(87)90581-2.CrossRefPubMed
7.
Kim HJ, Cui X, Hilsenbeck SG, Lee AV: Progesterone receptor loss correlates with human epidermal growth factor receptor 2 overexpression in estrogen receptor-positive breast cancer. Clin Cancer Res. 2006, 12 (3 Pt 2): 1013s-1018s.CrossRefPubMed
8.
Massarweh S, Schiff R: Resistance to endocrine therapy in breast cancer: exploiting estrogen receptor/growth factor signaling crosstalk. Endocr Relat Cancer. 2006, 13 (Suppl 1): S15-S24.CrossRefPubMed
9.
Massarweh S, Schiff R: Unraveling the mechanisms of endocrine resistance in breast cancer: new therapeutic opportunities. Clin Cancer Res. 2007, 13: 1950-1954. 10.1158/1078-0432.CCR-06-2540.CrossRefPubMed
10.
Konecny G, Pauletti G, Pegram M, Untch M, Dandekar S, Aguilar Z, Wilson C, Rong HM, Bauerfeind I, Felber M, Wang HJ, Beryt M, Seshadri R, Hepp H, Slamon DJ: Quantitative association between HER-2/neu and steroid hormone receptors in hormone receptor-positive primary breast cancer. J Natl Cancer Inst. 2003, 95: 142-153. 10.1093/jnci/95.2.142.CrossRefPubMed
11.
12.
Foukakis T, Astrom G, Lindstrom L, Hatschek T, Bergh J: When to order a biopsy to characterise a metastatic relapse in breast cancer. Ann Oncol. 2012, 23 (10): 349-353.CrossRef
13.
Lindstrom LS, Karlsson E, Wilking UM, Johansson U, Hartman J, Lidbrink EK, Hatschek T, Skoog L, Bergh J: Clinically used breast cancer markers such as estrogen receptor, progesterone receptor, and human epidermal growth factor receptor 2 Are unstable throughout tumor progression. J Clin Oncol. 2012, 30: 2601-2608. 10.1200/JCO.2011.37.2482.CrossRefPubMed
14.
Dowsett M, Cuzick J, Ingle J, Coates A, Forbes J, Bliss J, Buyse M, Baum M, Buzdar A, Colleoni M, Coombes C, Snowdon C, Gnant M, Jakesz R, Kaufmann M, Boccardo F, Godwin J, Davies C, Peto R: Meta-analysis of breast cancer outcomes in adjuvant trials of aromatase inhibitors versus tamoxifen. J Clin Oncol. 2010, 28: 509-518. 10.1200/JCO.2009.23.1274.CrossRefPubMed
15.
Gibson L, Lawrence D, Dawson C, Bliss J: Aromatase inhibitors for treatment of advanced breast cancer in postmenopausal women. Cochrane Database Syst Rev. 1996, John Wiley & Sons, Ltd
16.
Song RX, Santen RJ, Kumar R, Adam L, Jeng MH, Masamura S, Yue W: Adaptive mechanisms induced by long-term estrogen deprivation in breast cancer cells. Mol Cell Endocrinol. 2002, 193: 29-42. 10.1016/S0303-7207(02)00093-X.CrossRefPubMed
17.
Martin LA, Farmer I, Johnston SR, Ali S, Marshall C, Dowsett M: Enhanced estrogen receptor (ER) alpha, ERBB2, and MAPK signal transduction pathways operate during the adaptation of MCF-7 cells to long term estrogen deprivation. J Biol Chem. 2003, 278: 30458-30468. 10.1074/jbc.M305226200.CrossRefPubMed
18.
Yue W, Wang JP, Conaway MR, Li Y, Santen RJ: Adaptive hypersensitivity following long-term estrogen deprivation: involvement of multiple signal pathways. J Steroid Biochem Mol Biol. 2003, 86: 265-274. 10.1016/S0960-0760(03)00366-2.CrossRefPubMed
19.
Yue W, Wang JP, Conaway M, Masamura S, Li Y, Santen RJ: Activation of the MAPK pathway enhances sensitivity of MCF-7 breast cancer cells to the mitogenic effect of estradiol. Endocrinology. 2002, 143: 3221-3229. 10.1210/en.2002-220186.CrossRefPubMed
20.
Yue W, Fan P, Wang J, Li Y, Santen RJ: Mechanisms of acquired resistance to endocrine therapy in hormone-dependent breast cancer cells. J Steroid Biochem Mol Biol. 2007, 106: 102-110. 10.1016/j.jsbmb.2007.05.008.CrossRefPubMedPubMedCentral
21.
Al Saleh S, Al Mulla F, Luqmani YA: Estrogen receptor silencing induces epithelial to mesenchymal transition in human breast cancer cells. PLoS One. 2011, 6: e20610-10.1371/journal.pone.0020610.CrossRefPubMedPubMedCentral
22.
Chan CM, Martin L-A, Johnston SR, Ali S, Dowsett M: Molecular changes associated with the acquisition of oestrogen hypersensitivity in MCF-7 breast cancer cells on long-term oestrogen deprivation. J Steroid Biochem Mol Biol. 2002, 81: 333-341. 10.1016/S0960-0760(02)00074-2.CrossRefPubMed
23.
Miller TW, Hennessy BT, González-Angulo AM, Fox EM, Mills GB, Chen H, Higham C, García-Echeverría C, Shyr Y, Arteaga CL: Hyperactivation of phosphatidylinositol-3 kinase promotes escape from hormone dependence in estrogen receptor–positive human breast cancer. J Clin Investig. 2010, 120: 2406-2413. 10.1172/JCI41680.CrossRefPubMedPubMedCentral
24.
Aguilar H, Solé X, Bonifaci N, Serra-Musach J, Islam A, López-Bigas N, Méndez-Pertuz M, Beijersbergen RL, Lázaro C, Urruticoechea A, Pujana MA: Biological reprogramming in acquired resistance to endocrine therapy of breast cancer. Oncogene. 2010, 29: 6071-6083. 10.1038/onc.2010.333.CrossRefPubMed
25.
Hatzikirou H, Basanta D, Simon M, Schaller K, Deutsch A: “Go or grow”: the key to the emergence of invasion in tumour progression?. Math Med Biol. 2010, 29: 49-65.CrossRefPubMed
26.
Miller WR, Larionov AA, Renshaw L, Anderson TJ, White S, Murray J, Murray E, Hampton G, Walker JR, Ho S, Krause A, Evans DB, Dixon JM: Changes in breast cancer transcriptional profiles after treatment with the aromatase inhibitor, letrozole. Pharmacogenet Genomics. 2007, 17: 813-826. 10.1097/FPC.0b013e32820b853a.CrossRefPubMed
27.
Jeng MH, Shupnik MA, Bender TP, Westin EH, Bandyopadhyay D, Kumar R, Masamura S, Santen RJ: Estrogen receptor expression and function in long-term estrogen-deprived human breast cancer cells. Endocrinology. 1998, 139: 4164-4174. 10.1210/en.139.10.4164.PubMed
28.
Magnani L, Stoeck A, Zhang X, Lánczky A, Mirabella AC, Wang T-L, Gyorffy B, Lupien M: Genome-wide reprogramming of the chromatin landscape underlies endocrine therapy resistance in breast cancer. Proc Natl Acad Sci USA. 2013, 110: E1490-E1499. 10.1073/pnas.1219992110.CrossRefPubMedPubMedCentral
29.
Amir E, Miller N, Geddie W, Freedman O, Kassam F, Simmons C, Oldfield M, Dranitsaris G, Tomlinson G, Laupacis A, Tannock IF, Clemons M: Prospective study evaluating the impact of tissue confirmation of metastatic disease in patients with breast cancer. J Clin Oncol. 2011, 30: 587-592.CrossRefPubMed
30.
Thompson AM, Jordan LB, Quinlan P, Anderson E, Skene A, Dewar JA, Purdie CA: Prospective comparison of switches in biomarker status between primary and recurrent breast cancer: the breast recurrence in tissues study (BRITS). Breast Cancer Res. 2010, 12: R92-10.1186/bcr2771.CrossRefPubMedPubMedCentral
31.
Niikura N, Liu J, Hayashi N, Mittendorf EA, Gong Y, Palla SL, Tokuda Y, Gonzalez-Angulo AM, Hortobagyi GN, Ueno NT: Loss of human epidermal growth factor receptor 2 (HER2) expression in metastatic sites of HER2-overexpressing primary breast tumors. J Clin Oncol. 2011, 30: 593-599.CrossRefPubMedPubMedCentral
32.
Weigel MT, Ghazoui Z, Dunbier A, Pancholi S, Dowsett M, Martin LA: Preclinical and clinical studies of estrogen deprivation support the PDGF/Abl pathway as a novel therapeutic target for overcoming endocrine resistance in breast cancer. Breast Cancer Res. 2012, 14: R78-10.1186/bcr3191.CrossRefPubMedPubMedCentral
33.
Chen J, Imanaka N, Chen J, Griffin JD: Hypoxia potentiates notch signaling in breast cancer leading to decreased E-cadherin expression and increased cell migration and invasion. Br J Cancer. 2009, 102: 351-360.CrossRefPubMedPubMedCentral
34.
Bolos V, Mira E, Poveda BM, Luxan G, Canamero M, Martinez-A C, Manes S, de la Pompa JL: Notch activation stimulates migration of breast cancer cells and promotes tumor growth. Breast Cancer Res. 2013, 15: R54-10.1186/bcr3447.CrossRefPubMedPubMedCentral
35.
Scherbakov AM, Andreeva OE, Shatskaya VA, Krasil’nikov MA: The relationships between snail1 and estrogen receptor signaling in breast cancer cells. J Cell Biochem. 2012, 113: 2147-2155. 10.1002/jcb.24087.CrossRefPubMed
36.
Cui X, Schiff R, Arpino G, Osborne CK, Lee AV: Biology of progesterone receptor loss in breast cancer and its implications for endocrine therapy. J Clin Oncol. 2005, 23: 7721-7735. 10.1200/JCO.2005.09.004.CrossRefPubMed
37.
Cui X, Zhang P, Deng W, Oesterreich S, Lu Y, Mills GB, Lee AV: Insulin-like growth factor-I inhibits progesterone receptor expression in breast cancer cells via the phosphatidylinositol 3-kinase/Akt/mammalian target of rapamycin pathway: progesterone receptor as a potential indicator of growth factor activity in breast cancer. Mol Endocrinol. 2003, 17: 575-588. 10.1210/me.2002-0318.CrossRefPubMed
38.
Dowsett M, Harper-Wynne C, Boeddinghaus I, Salter J, Hills M, Dixon M, Ebbs S, Gui G, Sacks N, Smith I: HER-2 amplification impedes the antiproliferative effects of hormone therapy in estrogen receptor-positive primary breast cancer. Cancer Res. 2001, 61: 8452-8458.PubMed
39.
Oesterreich S, Zhang P, Guler RL, Sun X, Curran EM, Welshons WV, Osborne CK, Lee AV: Re-expression of estrogen receptor alpha in estrogen receptor alpha-negative MCF-7 cells restores both estrogen and insulin-like growth factor-mediated signaling and growth. Cancer Res. 2001, 61: 5771-5777.PubMed
40.
McClelland RA, Barrow D, Madden TA, Dutkowski CM, Pamment J, Knowlden JM, Gee JM, Nicholson RI: Enhanced epidermal growth factor receptor signaling in MCF7 breast cancer cells after long-term culture in the presence of the pure antiestrogen ICI 182,780 (faslodex). Endocrinology. 2001, 142: 2776-2788. 10.1210/en.142.7.2776.PubMed
Metadata
Title
Clinical instability of breast cancer markers is reflected in long-term in vitro estrogen deprivation studies
Authors
Jelena Milosevic
Johanna Klinge
Anna-Lena Borg
Theodoros Foukakis
Jonas Bergh
Nicholas P Tobin
Publication date
01-12-2013
Publisher
BioMed Central
Published in
BMC Cancer / Issue 1/2013
Electronic ISSN: 1471-2407
DOI
https://doi.org/10.1186/1471-2407-13-473

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