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Breast Cancer Research
Published in: Breast Cancer Research 5/2012

Open Access 01-10-2012 | Research article

Ets-1 is a transcriptional mediator of oncogenic nitric oxide signaling in estrogen receptor-negative breast cancer

Authors: Christopher H Switzer, Robert Y-S Cheng, Lisa A Ridnour, Sharon A Glynn, Stefan Ambs, David A Wink

Published in: Breast Cancer Research | Issue 5/2012

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Abstract

Introduction

The Ets-1 transcription factor is a candidate breast cancer oncogene that regulates the expression of genes involved in tumor progression and metastasis. Ets-1 signaling has also been linked to the development of a basal-like breast cancer phenotype. We recently described a nitric oxide (NO)-induced gene signature that is associated with poor disease outcome in estrogen receptor-negative (ER-) breast cancer and contains both stem cell-like and basal-like components. Thus, we examined the role of Ets-1 in NO signaling and NO-induced phenotypes in ER- human breast cancer cells.

Methods

Promoter region analyses were performed on genes upregulated in inducible nitric oxide synthase (NOS2) high expressing tumors for Ets-binding sites. In vitro mechanisms were examined in human basal-like breast cancer cells lines. NO signaling effects were studied using either forced NOS2 expression or the use of a chemical NO-donor, diethlylenetriamine NONOate (DETANO).

Results

Promoter region analysis of genes that are up-regulated in human ER-negative breast tumors with high NOS2 expression revealed that the Ets-binding sequence is the only common promoter element present in all of these genes, indicating that Ets-1 is the key transcriptional factor down-stream of oncogenic NOS2-signaling. Accordingly, both forced NOS2 over-expression and exposure to NO-donors resulted in significant Ets-1 transcriptional activation in ER- breast cancer cells. Functional studies showed that NO activated Ets-1 transcriptional activity via a Ras/MEK/ERK signaling pathway by a mechanism that involved Ras S-nitrosylation. RNA knock-down of Ets-1 suppressed NO-induced expression of selected basal-like breast cancer markers such as P-cadherin, S100A8, IL-8 and αβ-crystallin. Additionally, Ets-1 knock-down reduced NO-mediated cellular proliferation, matrix metalloproteinase and cathepsin B activities, as well as matrigel invasion.

Conclusions

These data show that Ets-1 is a key transcriptional mediator of oncogenic NO signaling that promotes the development of an aggressive disease phenotype in ER- breast cancer in an Ets-1 and Ras-dependent manner, providing novel clues of how NOS2 expression in human breast tumors is functionally linked to poor patient survival.
Appendix
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Literature
1.
Wink DA, Hines HB, Cheng RYS, Switzer CH, Flores-Santana W, Vitek MP, Ridnour LA, Colton CA: Nitric oxide and redox mechanisms in the immune response. J Leukoc Biol. 2011, 89: 873-891. 10.1189/jlb.1010550.CrossRefPubMedPubMedCentral
2.
Eyler CE, Wu Q, Yan K, MacSwords JM, Chandler-Militello D, Misuraca KL, Lathia JD, Forrester MT, Lee J, Stamler JS, Goldman SA, Bredel M, McLendon RE, Sloan AE, Hjelmeland AB, Rich JN: Glioma stem cell proliferation and tumor growth are promoted by nitric oxide synthase-2. Cell. 2011, 146: 53-66. 10.1016/j.cell.2011.06.006.CrossRefPubMedPubMedCentral
3.
Cheng R, Ridnour LA, Glynn SA, Switzer CH, Flores-Santana W, Hussain P, Thomas DD, Ambs S, Harris CC, Wink DA: Nitric Oxide and Cancer: An Overview. Nitric Oxide (NO) and Cancer. Edited by: Bonavida B. 2010, New York: Springer, 3-20.CrossRef
4.
Ekmekcioglu S, Tang CH, Grimm EA: NO news is not necessarily good news in cancer. Curr Cancer Drug Targets. 2005, 5: 103-115. 10.2174/1568009053202072.CrossRefPubMed
5.
Glynn SA, Boersma BJ, Dorsey TH, Yi M, Yfantis HG, Ridnour LA, Martin DN, Switzer CH, Hudson RS, Wink DA, Lee DH, Stephens RM, Ambs S: Increased NOS2 predicts poor survival in estrogen receptor-negative breast cancer patients. J Clin Invest. 2010, 120: 3843-3854. 10.1172/JCI42059.CrossRefPubMedPubMedCentral
6.
Reis-Filho JS, Tutt ANJ: Triple negative tumours: a critical review. Histopathology. 2008, 52: 108-118.CrossRefPubMed
7.
Rakha EA, Reis-Filho JS, Ellis IO: Basal-like breast cancer: a critical review. J Clin Oncol. 2008, 26: 2568-2581. 10.1200/JCO.2007.13.1748.CrossRefPubMed
8.
Switzer CH, Glynn SA, Ridnour LA, Cheng RYS, Vitek MP, Ambs S, Wink DA: Nitric oxide and protein phosphatase 2A provide novel therapeutic opportunities in ER-negative breast cancer. Trends Pharmacol Sci. 2011, 32: 644-651. 10.1016/j.tips.2011.07.001.CrossRefPubMedPubMedCentral
9.
Thomas DD, Ridnour LA, Isenberg JS, Flores-Santana W, Switzer CH, Donzelli S, Hussain P, Vecoli C, Paolocci N, Ambs S, Colton CA, Harris CC, Roberts DD, Wink DA: The chemical biology of nitric oxide: implications in cellular signaling. Free Radic Biol Med. 2008, 45: 18-31. 10.1016/j.freeradbiomed.2008.03.020.CrossRefPubMedPubMedCentral
10.
11.
Fukumura D, Kashiwagi S, Jain RK: The role of nitric oxide in tumour progression. Nat Rev Cancer. 2006, 6: 521-534. 10.1038/nrc1910.CrossRefPubMed
12.
Prueitt RL, Boersma BJ, Howe TM, Goodman JE, Thomas DD, Ying L, Pfiester CM, Yfantis HG, Cottrell JR, Lee DH, Remaley AT, Hofseth LJ, Wink DA, Ambs S: Inflammation and IGF-I activate the Akt pathway in breast cancer. Int J Cancer. 2007, 120: 796-805. 10.1002/ijc.22336.CrossRefPubMed
13.
Thomas DD, Espey MG, Ridnour LA, Hofseth LJ, Mancardi D, Harris CC, Wink DA: Hypoxic inducible factor 1a, extracellular signal-regulated kinase, and p53 are regulated by distinct threshold concentrations of nitric oxide. Proc Natl Acad Sci USA. 2004, 101: 8894-8899. 10.1073/pnas.0400453101.CrossRefPubMedPubMedCentral
14.
Rahman MA, Senga T, Ito S, Hyodo T, Hasegawa H, Hamaguchi M: S-nitrosylation at cysteine 498 of c-Src tyrosine kinase regulates nitric oxide-mediated cell invasion. J Biol Chem. 2010, 285: 3806-3814. 10.1074/jbc.M109.059782.CrossRefPubMed
15.
Lee H-C, An S, Lee H, Woo S-H, Jin H-O, Seo S-K, Choe T-B, Yoo D-H, Lee S-J, Hong Y-J, Park M-J, Rhee C-H, Park I-C, Hong S-I: Activation of epidermal growth factor receptor and its downstream signaling pathway by nitric oxide in response to ionizing radiation. Mol Cancer Res. 2008, 6: 996-1002. 10.1158/1541-7786.MCR-08-0113.CrossRefPubMed
16.
Hahne JC, Okuducu AF, Kaminski A, Florin A, Soncin F, Wernert N: Ets-1 expression promotes epithelial cell transformation by inducing migration, invasion and anchorage-independent growth. Oncogene. 2005, 24: 5384-5388. 10.1038/sj.onc.1208761.CrossRefPubMed
17.
Furlan A, Vercamer C, Desbiens X, Pourtier A: Ets-1 triggers and orchestrates the malignant phenotype of mammary cancer cells within their matrix environment. J Cell Physiol. 2008, 215: 782-793. 10.1002/jcp.21360.CrossRefPubMed
18.
Turner DP, Findlay VJ, Moussa O, Watson DK: Defining ETS transcription regulatory networks and their contribution to breast cancer progression. J Cell Biochem. 2007, 102: 549-559. 10.1002/jcb.21494.CrossRefPubMed
19.
Mylona EE, Alexandrou PT, Giannopoulou IA, Rafailidis PI, Markaki S, Keramopoulos A, Nakopoulou LL: Study of the topographic distribution of ets-1 protein expression in invasive breast carcinomas in relation to tumor phenotype. Cancer Detect Prev. 2006, 30: 111-117. 10.1016/j.cdp.2006.03.006.CrossRefPubMed
20.
Lincoln DW 2nd, Bove K: The transcription factor Ets-1 in breast cancer. Front Biosci. 2005, 10: 506-511. 10.2741/1546.CrossRefPubMed
21.
Buggy Y, Maguire TM, McGreal G, McDermott E, Hill AD, O'Higgins N, Duffy MJ: Overexpression of the Ets-1 transcription factor in human breast cancer. Br J Cancer. 2004, 91: 1308-1315. 10.1038/sj.bjc.6602128.CrossRefPubMedPubMedCentral
22.
Span PN, Manders P, Heuvel JJ, Thomas CM, Bosch RR, Beex LV, Sweep CG: Expression of the transcription factor Ets-1 is an independent prognostic marker for relapse-free survival in breast cancer. Oncogene. 2002, 21: 8506-8509. 10.1038/sj.onc.1206040.CrossRefPubMed
23.
Bosman JD, Yehiely F, Evans JR, Cryns VL: Regulation of alphaB-crystallin gene expression by the transcription factor Ets1 in breast cancer. Breast Cancer Res Treat. 2010, 119: 63-70. 10.1007/s10549-009-0330-4.CrossRefPubMed
24.
Yang BS, Hauser CA, Henkel G, Colman MS, Van Beveren C, Stacey KJ, Hume DA, Maki RA, Ostrowski MC: Ras-mediated phosphorylation of a conserved threonine residue enhances the transactivation activities of c-Ets1 and c-Ets2. Mol Cell Biol. 1996, 16: 538-547.CrossRefPubMedPubMedCentral
25.
Wasylyk C, Bradford AP, Gutierrez-Hartmann A, Wasylyk B: Conserved mechanisms of Ras regulation of evolutionary related transcription factors, Ets1 and Pointed P2. Oncogene. 1997, 14: 899-913. 10.1038/sj.onc.1200914.CrossRefPubMed
26.
Paumelle R, Tulasne D, Kherrouche Z, Plaza S, Leroy C, Reveneau S, Vandenbunder B, Fafeur V: Hepatocyte growth factor/scatter factor activates the ETS1 transcription factor by a RAS-RAF-MEK-ERK signaling pathway. Oncogene. 2002, 21: 2309-2319. 10.1038/sj.onc.1205297.CrossRefPubMed
27.
Iwasaka C, Tanaka K, Abe M, Sato Y: Ets-1 regulates angiogenesis by inducing the expression of urokinase-type plasminogen activator and matrix metalloproteinase-1 and the migration of vascular endothelial cells. J Cell Physiol. 1996, 169: 522-531. 10.1002/(SICI)1097-4652(199612)169:3<522::AID-JCP12>3.0.CO;2-7.CrossRefPubMed
28.
Chen Z, Fisher RJ, Riggs CW, Rhim JS, Lautenberger JA: Inhibition of vascular endothelial growth factor-induced endothelial cell migration by ETS1 antisense oligonucleotides. Cancer Res. 1997, 57: 2013-2019.PubMed
29.
Sahin A, Velten M, Pietsch T, Knuefermann P, Okuducu AF, Hahne JC, Wernert N: Inactivation of Ets 1 transcription factor by a specific decoy strategy reduces rat C6 glioma cell proliferation and mmp-9 expression. Int J Mol Med. 2005, 15: 771-776.PubMed
30.
Ridnour LA, Windhausen AN, Isenberg JS, Yeung N, Thomas DD, Vitek MP, Roberts DD, Wink DA: Nitric oxide regulates matrix metalloproteinase-9 activity by guanylyl-cyclase-dependent and -independent pathways. Proc Natl Acad Sci USA. 2007, 104: 16898-16903. 10.1073/pnas.0702761104.CrossRefPubMedPubMedCentral
31.
Keefer LK, Nims RW, Davies KM, Wink DA: "NONOates" (1-substituted diazen-1-ium-1,2-diolates) as nitric oxide donors: convenient nitric oxide dosage forms. Methods Enzymol. 1996, 268: 281-293.CrossRefPubMed
32.
Kolch W: Meaningful relationships: the regulation of the Ras/Raf/MEK/ERK pathway by protein interactions. Biochem J. 2000, 351: 289-305. 10.1042/0264-6021:3510289.CrossRefPubMedPubMedCentral
33.
Lander HM, Hajjar DP, Hempstead BL, Mirza UA, Chait BT, Campbell S, Quilliam LA: A molecular redox switch on p21(ras). Structural basis for the nitric oxide-p21(ras) interaction. J Biol Chem. 1997, 272: 4323-4326. 10.1074/jbc.272.7.4323.CrossRefPubMed
34.
Mirza UA, Chait BT, Lander HM: Monitoring reactions of nitric oxide with peptides and proteins by electrospray ionization-mass spectrometry. J Biol Chem. 1995, 270: 17185-17188. 10.1074/jbc.270.29.17185.CrossRefPubMed
35.
Jaffrey SR, Snyder SH: The biotin switch method for the detection of S-nitrosylated proteins. Sci STKE. 2001, 2001: pl1-PubMed
36.
Wink DA, Nims RW, Darbyshire JF, Christodoulou D, Hanbauer I, Cox GW, Laval F, Laval J, Cook JA, Krishna MC: Reaction kinetics for nitrosation of cysteine and glutathione in aerobic nitric oxide solutions at neutral pH. Insights into the fate and physiological effects of intermediates generated in the NO/O2 reaction. Chem Res Toxicol. 1994, 7: 519-525. 10.1021/tx00040a007.CrossRefPubMed
37.
Haklai R, Weisz MG, Elad G, Paz A, Marciano D, Egozi Y, Ben-Baruch G, Kloog Y: Dislodgment and accelerated degradation of Ras. Biochemistry. 1998, 37: 1306-1314. 10.1021/bi972032d.CrossRefPubMed
38.
Lindemann RK, Braig M, Ballschmieter P, Guise TA, Nordheim A, Dittmer J: Protein kinase Calpha regulates Ets1 transcriptional activity in invasive breast cancer cells. Int J Oncol. 2003, 22: 799-805.PubMed
39.
Kolch W, Heidecker G, Kochs G, Hummel R, Vahidi H, Mischak H, Finkenzeller G, Marme D, Rapp UR: Protein kinase C[alpha] activates RAF-1 by direct phosphorylation. Nature. 1993, 364: 249-252. 10.1038/364249a0.CrossRefPubMed
40.
Vetter M, Blumenthal SG, Lindemann RK, Manns J, Wesselborg S, Thomssen C, Dittmer J: Ets1 is an effector of protein kinase C[alpha] in cancer cells. Oncogene. 2005, 24: 650-661. 10.1038/sj.onc.1208234.CrossRefPubMed
41.
42.
Behrens P, Rothe M, Wellmann A, Krischler J, Wernert N: The Ets-1 transcription factor is up-regulated together with MMP 1 and MMP 9 in the stroma of pre-invasive breast cancer. J Pathol. 2001, 194: 43-50. 10.1002/path.844.CrossRefPubMed
43.
Sasaki H, Yukiue H, Moiriyama S, Kobayashi Y, Nakashima Y, Kaji M, Kiriyama M, Fukai I, Yamakawa Y, Fujii Y: Clinical significance of matrix metalloproteinase-7 and Ets-1 gene expression in patients with lung cancer. J Surg Res. 2001, 101: 242-247. 10.1006/jsre.2001.6279.CrossRefPubMed
44.
Duffy MJ, Maguire TM, Hill A, McDermott E, O'Higgins N: Metalloproteinases: role in breast carcinogenesis, invasion and metastasis. Breast Cancer Res. 2000, 2: 252-257. 10.1186/bcr65.CrossRefPubMedPubMedCentral
45.
Sørlie T, Perou CM, Tibshirani R, Aas T, Geisler S, Johnsen H, Hastie T, Eisen MB, van de Rijn M, Jeffrey SS, Thorsen T, Quist H, Matese JC, Brown PO, Botstein D, Lønning PE, Børresen-Dale A-L: Gene expression patterns of breast carcinomas distinguish tumor subclasses with clinical implications. Proc Natl Acad Sci USA. 2001, 98: 10869-10874. 10.1073/pnas.191367098.CrossRefPubMedPubMedCentral
46.
Charafe-Jauffret E, Ginestier C, Monville F, Finetti P, Adelaide J, Cervera N, Fekairi S, Xerri L, Jacquemier J, Birnbaum D, Bertucci F: Gene expression profiling of breast cell lines identifies potential new basal markers. Oncogene. 2006, 25: 2273-2284. 10.1038/sj.onc.1209254.CrossRefPubMed
47.
Rody A, Karn T, Liedtke C, Pusztai L, Ruckhaeberle E, Hanker L, Gaetje R, Solbach C, Ahr A, Metzler D, Schmidt M, Muller V, Holtrich U, Kaufmann M: A clinically relevant gene signature in triple negative and basal-like breast cancer. Breast Cancer Res. 2011, 13: R97-10.1186/bcr3035.CrossRefPubMedPubMedCentral
48.
Sitterding SM, Wiseman WR, Schiller CL, Luan C, Chen F, Moyano JV, Watkin WG, Wiley EL, Cryns VL, Diaz LK: AlphaB-crystallin: a novel marker of invasive basal-like and metaplastic breast carcinomas. Ann Diagn Pathol. 2008, 12: 33-40. 10.1016/j.anndiagpath.2007.02.004.CrossRefPubMed
49.
Withana NP, Blum G, Sameni M, Slaney C, Anbalagan A, Olive MB, Bidwell BN, Edgington L, Wang L, Moin K, Sloane BF, Anderson RL, Bogyo MS, Parker BS: Cathepsin B inhibition limits bone metastasis in breast cancer. Cancer Res. 2012, 72: 1199-1209. 10.1158/0008-5472.CAN-11-2759.CrossRefPubMedPubMedCentral
50.
Nomura T, Katunuma N: Involvement of cathepsins in the invasion, metastasis and proliferation of cancer cells. J Med Invest. 2005, 52: 1-9. 10.2152/jmi.52.1.CrossRefPubMed
51.
Ridnour LA, Thomas DD, Switzer C, Flores-Santana W, Isenberg JS, Ambs S, Roberts DD, Wink DA: Molecular mechanisms for discrete nitric oxide levels in cancer. Nitric Oxide. 2008, 19: 73-76. 10.1016/j.niox.2008.04.006.CrossRefPubMedPubMedCentral
52.
von Lintig FC, Dreilinger AD, Varki NM, Wallace AM, Casteel DE, Boss GR: Ras activation in human breast cancer. Breast Cancer Res Treat. 2000, 62: 51-62. 10.1023/A:1006491619920.CrossRefPubMed
53.
Sanchez-Munoz A, Gallego E, de Luque V, Perez-Rivas LG, Vicioso L, Ribelles N, Lozano J, Alba E: Lack of evidence for KRAS oncogenic mutations in triple-negative breast cancer. BMC Cancer. 2010, 10: 136-10.1186/1471-2407-10-136.CrossRefPubMedPubMedCentral
54.
Deora AA, Hajjar DP, Lander HM: Recruitment and activation of Raf-1 kinase by nitric oxide-activated Ras. Biochemistry. 2000, 39: 9901-9908. 10.1021/bi992954b.CrossRefPubMed
55.
Marozkina NV, Wei C, Yemen S, Wallrabe H, Nagji AS, Liu L, Morozkina T, Jones DR, Gaston B: S-Nitrosoglutathione reductase in human lung cancer. Am J Respir Cell Mol Biol. 2012, 46: 63-70. 10.1165/rcmb.2011-0147OC.CrossRefPubMedPubMedCentral
56.
Pervin S, Singh R, Hernandez E, Wu G, Chaudhuri G: Nitric oxide in physiologic concentrations targets the translational machinery to increase the proliferation of human breast cancer cells: involvement of mammalian target of rapamycin/eIF4E pathway. Cancer Res. 2007, 67: 289-299. 10.1158/0008-5472.CAN-05-4623.CrossRefPubMed
57.
Ekmekcioglu S, Ellerhorst J, Smid CM, Prieto VG, Munsell M, Buzaid AC, Grimm EA: Inducible nitric oxide synthase and nitrotyrosine in human metastatic melanoma tumors correlate with poor survival. Clin Cancer Res. 2000, 6: 4768-4775.PubMed
58.
Wei W, Li B, Hanes MA, Kakar S, Chen X, Liu L: S-Nitrosylation from GSNOR deficiency impairs DNA repair and promotes hepatocarcinogenesis. Sci Transl Med. 2: 19ra13-
59.
Yamaguchi E, Nakayama T, Nanashima A, Matsumoto K, Yasutake T, Sekine I, Nagayasu T: Ets-1 proto-oncogene as a potential predictor for poor prognosis of lung adenocarcinoma. Tohoku J Exp Med. 2007, 213: 41-50. 10.1620/tjem.213.41.CrossRefPubMed
60.
Rothhammer T, Hahne JC, Florin A, Poser I, Soncin F, Wernert N, Bosserhoff AK: The Ets-1 transcription factor is involved in the development and invasion of malignant melanoma. Cell Mol Life Sci. 2004, 61: 118-128. 10.1007/s00018-003-3337-8.CrossRefPubMed
Metadata
Title
Ets-1 is a transcriptional mediator of oncogenic nitric oxide signaling in estrogen receptor-negative breast cancer
Authors
Christopher H Switzer
Robert Y-S Cheng
Lisa A Ridnour
Sharon A Glynn
Stefan Ambs
David A Wink
Publication date
01-10-2012
Publisher
BioMed Central
Published in
Breast Cancer Research / Issue 5/2012
Electronic ISSN: 1465-542X
DOI
https://doi.org/10.1186/bcr3319

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