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BMC Cancer
Published in: BMC Cancer 1/2016

Open Access 01-12-2016 | Research article

uPAR enhances malignant potential of triple-negative breast cancer by directly interacting with uPA and IGF1R

Authors: Michaela C. Huber, Rebecca Mall, Herbert Braselmann, Annette Feuchtinger, Sara Molatore, Katrin Lindner, Axel Walch, Eva Gross, Manfred Schmitt, Natalie Falkenberg, Michaela Aubele

Published in: BMC Cancer | Issue 1/2016

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Abstract

Background

Due to lack of a targeted therapy for the triple-negative breast cancer (TNBC) patients, it is important to explore this aggressive breast cancer type in more detail and to establish novel therapeutic approaches. TNBC is defined negative for the protein expression of oestrogen receptor (ER), progesterone receptor (PR) and human epidermal growth factor receptor 2 (HER2). One prominent feature of this cancer type is the frequent overexpression of major components of the urokinase-type plasminogen activator system (uPAS) including uPA, its receptor uPAR and the inhibitor PAI-1, which may be valuable as therapeutic targets.

Methods

Direct interactions of uPAR with interactors were demonstrated by immunoprecipitations and proximity ligation assays. For stable knockdowns of target proteins, lentiviral vectors were used and the effects were analysed by immunoblottings and using in vitro cell viability, migration and invasion assays. Immunohistochemical and statistical analyses of biomarkers and clinical parameters were conducted in a TNBC cohort (n = 174).

Results

Direct tumour-promoting interactions of uPAR with uPA and the insulin-like growth factor receptor 1 (IGF1R) were shown in TNBC cells and these interactions were significantly reduced (p = 0.001) when uPAR was downregulated. The combined knockdown of uPAR and uPA or IGF1R additively and significantly reduced cell viability, migration and invasion of the model cell lines. In TNBC tissue, the complexes formed by uPAR with uPA or with IGF1R significantly correlated with the histological grade (p = 0.0019) as well as with cathepsin B and D (p ≤ 0.0001) that are implicated in cell invasion and metastasis.

Conclusions

Our outcomes show that not only overexpressed biomarkers promote tumourigenesis, but rather their interactions further potentiate tumour progression. This study emphasises the potential of combined approaches targeting uPAR and its interactors with regard to an improved therapy of TNBC.
Appendix
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Literature
1.
2.
Morris GJ, Naidu S, Topham AK, Guiles F, Xu YH, McCue P, et al. Differences in breast carcinoma characteristics in newly diagnosed African-American and Caucasian patients - A single-institution compilation compared with the National Cancer Institute’s Surveillance, Epidemiology, and End Results Database. Cancer. 2007;110:876–84.CrossRefPubMed
3.
Singh P, Alex JM, Bast F. Insulin receptor (IR) and insulin-like growth factor receptor 1 (IGF-1R) signaling systems: novel treatment strategies for cancer. Med Oncol. 2014;31:1.CrossRef
4.
Davison Z, de Blacquiere GE, Westley BR, May FEB. Insulin-like growth factor-dependent proliferation and survival of triple-negative breast cancer cells: implications for therapy. Neoplasia. 2011;13:504–15.CrossRefPubMedPubMedCentral
5.
Nielsen TO, Hsu FD, Jensen K, Cheang M, Karaca G, Hu ZY, et al. Immunohistochemical and clinical characterization of the basal-like subtype of invasive breast carcinoma. Clin Cancer Res. 2004;10:5367–74.CrossRefPubMed
6.
Nielsen TO, Andrews HN, Cheang M, Kucab JE, Hsu FD, Ragaz J, et al. Expression of the insulin-like growth factor I receptor and urokinase plasminogen activator in breast cancer is associated with poor survival: Potential for intervention with 17-allylamino geldanamycin. Cancer Res. 2004;64:286–91.CrossRefPubMed
7.
Lengyel E, Prechtel D, Resau JH, Gauger K, Welk A, Lindemann K, et al. c-Met overexpression in node-positive breast cancer identifies patients with poor clinical outcome independent of Her2/neu. Int J Cancer. 2005;113:678–82.CrossRefPubMed
8.
Duffy MJ, McGowan PM, Crown J. Targeted therapy for triple-negative breast cancer: Where are we? Int J Cancer. 2012;131:2471–77.CrossRefPubMed
9.
Foekens JA, Peters HA, Look MP, Portengen H, Schmitt M, Kramer MD, et al. The urokinase system of plasminogen activation and prognosis in 2780 breast cancer patients. Cancer Res. 2000;60:636–43.PubMed
10.
Riedemann J, Macaulay VM. IGF1R signalling and its inhibition. Endocr Relat Cancer. 2006;13:S33–43.CrossRefPubMed
11.
Bonine-Summers AR, Aakre ME, Brown KA, Arteaga CL, Pietenpol JA, Moses HL, et al. Epidermal growth factor receptor plays a significant role in hepatocyte growth factor mediated biological responses in mammary epithelial cells. Cancer Biol Ther. 2007;6:561–70.CrossRefPubMedPubMedCentral
12.
Law JH, Habibi G, Hu K, Masoudi H, Wang MYC, Stratford AL, et al. Phosphorylated insulin-like growth factor-I/insulin receptor is present in all breast cancer subtypes and is related to poor survival. Cancer Res. 2008;68:10238–46.CrossRefPubMed
13.
Meyer AS, Miller MA, Gertler FB, Lauffenburger DA. The receptor AXL diversifies EGFR signaling and limits the response to EGFR-targeted inhibitors in triple-negative breast cancer cells. Sci Signal. 2013;6:287.CrossRef
14.
Blasi F, Carmeliet P. uPAR: A versatile signalling orchestrator. Nat Rev Mol Cell Bio. 2002;3:932–43.CrossRef
15.
Duffy MJ, Duggan C, Maguire T, Mulcahy K, Elvin P, McDermott E, et al. Urokinase plasminogen activator as a predictor of aggressive disease in breast cancer. Enzyme Protein. 1996;49:85–93.PubMed
16.
Ullrich A, Gray A, Tam AW, Yang-Feng T, Tsubokawa M, Collins C, et al. Insulin-like growth factor I receptor primary structure: comparison with insulin receptor suggests structural determinants that define functional specificity. EMBO J. 1986;5:2503–12.PubMedPubMedCentral
17.
Corkery B, Crown J, Clynes M, O’Donovan N. Epidermal growth factor receptor as a potential therapeutic target in triple-negative breast cancer. Ann Oncol. 2009;20:862–67.CrossRefPubMed
18.
Ferraro DA, Gaborit N, Maron R, Cohen-Dvashi H, Porat Z, Pareja F, et al. Inhibition of triple-negative breast cancer models by combinations of antibodies to EGFR. Proc Natl Acad Sci U S A. 2013;110:1815–20.CrossRefPubMedPubMedCentral
19.
Carey LA, Rugo HS, Marcom PK, Mayer EL, Esteva FJ, Ma CX, et al. TBCRC 001: randomized phase II study of cetuximab in combination with carboplatin in stage IV triple-negative breast cancer. J Clin Oncol. 2012;30:2615–23.CrossRefPubMedPubMedCentral
20.
Nabholtz JM, Abrial C, Mouret-Reynier MA, Dauplat MM, Weber B, Gligorov J, et al. Multicentric neoadjuvant phase II study of panitumumab combined with an anthracycline/taxane-based chemotherapy in operable triple-negative breast cancer: identification of biologically defined signatures predicting treatment impact. Ann Oncol. 2014;25:1570–7.CrossRefPubMed
21.
Bernsdorf M, Ingvar C, Jorgensen L, Tuxen MK, Jakobsen EH, Saetersdal A, et al. Effect of adding gefitinib to neoadjuvant chemotherapy in estrogen receptor negative early breast cancer in a randomized phase II trial. Breast Cancer Res Treat. 2011;126:463–70.CrossRefPubMed
22.
Layman RM, Ruppert AS, Lynn M, Mrozek E, Ramaswamy B, Lustberg MB, et al. Severe and prolonged lymphopenia observed in patients treated with bendamustine and erlotinib for metastatic triple negative breast cancer. Cancer Chemother Pharmacol. 2013;71:1183–90.CrossRefPubMedPubMedCentral
23.
Yap TA, Olmos D, Brunetto AT, Tunariu N, Barriuso J, Riisnaes R, et al. Phase I Trial of a Selective c-MET Inhibitor ARQ 197 Incorporating Proof of Mechanism Pharmacodynamic Studies. J Clin Oncol. 2011;29:1271–79.CrossRefPubMed
24.
Yakes FM, Chen J, Tan J, Yamaguchi K, Shi YC, Yu PW, et al. Cabozantinib (XL184), a Novel MET and VEGFR2 Inhibitor, Simultaneously Suppresses Metastasis, Angiogenesis, and Tumor Growth. Mol Cancer Ther. 2011;10:2298–308.CrossRefPubMed
25.
Ho-Yen CM, Jones JL, Kermorgant S. The clinical and functional significance of c-Met in breast cancer: a review. Breast Cancer Res. 2015;17.
26.
Hynes NE, Lane HA. ERBB receptors and cancer: The complexity of targeted inhibitors. Nat Rev Cancer. 2005;5:341–54.CrossRefPubMed
27.
Janicke F, Prechtl A, Thomssen C, Harbeck N, Meisner C, Untch M, et al. Randomized adjuvant chemotherapy trial in high-risk, lymph node-negative breast cancer patients identified by urokinase-type plasminogen activator and plasminogen activator inhibitor type 1. J Nat Cancer Inst. 2001;93:913–20.CrossRefPubMed
28.
Look MP, van Putten WLJ, Duffy MJ, Harbeck N, Christensen IJ, Thomssen C, et al. Pooled analysis of prognostic impact of urokinase-type plasminogen activator and its inhibitor PAI-1 8377 breast cancer patients. J Nat Cancer Inst. 2002;94:116–28.CrossRefPubMed
29.
Guo Y, Higazi AA, Arakelian A, Sachais BS, Cines D, Goldfarb RH, et al. A peptide derived from the nonreceptor binding region of urokinase plasminogen activator (uPA) inhibits tumor progression and angiogenesis and induces tumor cell death in vivo. FASEB J. 2000;14:1400–10.CrossRefPubMed
30.
Setyono-Han B, Sturzebecher J, Schmalix WA, Muehlenweg B, Sieuwerts AM, Timmermans M, et al. Suppression of rat breast cancer metastasis and reduction of primary tumour growth by the small synthetic urokinase inhibitor WX-UKI. Thromb Haemost. 2005;93:779–86.PubMed
31.
Subramanian R, Gondi CS, Lakka SS, Jutla A, Rao JS. siRNA-mediated simultaneous downregulation of uPA and its receptor inhibits angiogenesis and invasiveness triggering apoptosis in breast cancer cells. Int J Oncol. 2006;28:831–9.PubMedPubMedCentral
32.
Li CF, Cao S, Liu Z, Ye X, Chen LZ, Meng SD. RNAi-mediated downregulation of uPAR synergizes with targeting of HER2 through the ERK pathway in breast cancer cells. Int J Cancer. 2010;127:1507–16.CrossRefPubMed
33.
Kunigal S, Lakka SS, Good CS, Estes N, Rao JS. RNAi-mediated downregulation of urokinase plasminogen activator receptor and matrix metalloprotease-9 in human breast cancer cells results in decreased tumor invasion, angiogenesis and growth. Int J Cancer. 2007;121:2307–16.CrossRefPubMedPubMedCentral
34.
Bauer TW, Liu WB, Fan F, Camp ER, Yang A, Somcio RJ, et al. Targeting of urokinase plasminogen activator receptor in human pancreatic carcinoma cells inhibits c-met- and insulin-like growth factor-1 receptor-mediated migration and invasion and orthotopic tumor growth in mice. Cancer Res. 2005;65:7775–81.PubMed
35.
Bauer TW, Fan F, Liu WB, Johnson M, Parikh NU, Parry GC, et al. Insulinlike growth factor-I-mediated migration and invasion of human colon carcinoma cells requires activation of c-Met and urokinase plasminogen activator receptor. Ann Surg. 2005;241:748–58.CrossRefPubMedPubMedCentral
36.
Gallicchio MA, Kaun C, Wojta J, Binder B, Bach LA. Urokinase type plasminogen activator receptor is involved in insulin-like growth factor-induced migration of rhabdomyosarcoma cells in vitro. J Cell Physiol. 2003;197:131–8.CrossRefPubMed
37.
Ludyga N, Anastasov N, Rosemann M, Seiler J, Lohmann N, Braselmann H, et al. Effects of Simultaneous Knockdown of HER2 and PTK6 on Malignancy and Tumor Progression in Human Breast Cancer Cells. Mol Cancer Res. 2013;11:381–92.CrossRefPubMed
38.
D’Alessio S, Gerasi L, Blasi F. uPAR-deficient mouse keratinocytes fail to produce EGFR-dependent laminin-5, affecting migration in vivo and in vitro. J Cell Sci. 2008;121:3922–32.CrossRefPubMed
39.
Falkenberg N, Anastasov N, Hofig I, Bashkueva K, Lindner K, Hofler H, et al. Additive impact of HER2-/PTK6-RNAi on interactions with HER3 or IGF-1R leads to reduced breast cancer progression in vivo. Mol Oncol. 2015;9:282–94.CrossRefPubMed
40.
Falkenberg N, Anastasov N, Schaub A, Radulovic V, Schmitt M, Magdolen V, et al. Secreted uPAR isoform 2 (uPAR7b) is a novel direct target of miR-221. Oncotarget. 2015;6:8103–14.CrossRefPubMedPubMedCentral
41.
Luther T, Magdolen V, Albrecht S, Kasper M, Riemer C, Kessler H, et al. Epitope-mapped monoclonal antibodies as tools for functional and morphological analyses of the human urokinase receptor in tumor tissue. Am J Pathol. 1997;150:1231–44.PubMedPubMedCentral
42.
Ludyga N, Anastasov N, Gonzalez-Vasconcellos I, Ram M, Hofler H, Aubele M. Impact of protein tyrosine kinase 6 (PTK6) on human epidermal growth factor receptor (HER) signalling in breast cancer. Mol Biosyst. 2011;7:1603–12.CrossRefPubMed
43.
Molatore S, Kiermaier E, Jung CB, Lee M, Pulz E, Hofler H, et al. Characterization of a naturally-occurring p27 mutation predisposing to multiple endocrine tumors. Mol Cancer. 2010;9:116.CrossRefPubMedPubMedCentral
44.
Ludyga N, Grunwald B, Azimzadeh O, Englert S, Hofler H, Tapio S, et al. Nucleic acids from long-term preserved FFPE tissues are suitable for downstream analyses. Virchows Arch. 2012;460:131–40.CrossRefPubMed
45.
Elston CW, Ellis IO. Pathological prognostic factors in breast cancer. I. The value of histological grade in breast cancer: experience from a large study with long-term follow-up. Histopathology. 1991;19:403–10.CrossRefPubMed
46.
Aubele M, Auer G, Walch AK, Munro A, Atkinson MJ, Braselmann H, et al. PTK (protein tyrosine kinase)-6 and HER2 and 4, but not HER1 and 3 predict long-term survival in breast carcinomas. Br J Cancer. 2007;96:801–7.CrossRefPubMedPubMedCentral
47.
Aubele M, Spears M, Ludyga N, Braselmann H, Feuchtinger A, Taylor KJ, et al. In situ quantification of HER2-protein tyrosine kinase 6 (PTK6) protein-protein complexes in paraffin sections from breast cancer tissues. Br J Cancer. 2010;103:663–7.CrossRefPubMedPubMedCentral
48.
Lehmann BD, Bauer JA, Chen X, Sanders ME, Chakravarthy AB, Shyr Y, et al. Identification of human triple-negative breast cancer subtypes and preclinical models for selection of targeted therapies. J Clin Invest. 2011;121:2750–67.CrossRefPubMedPubMedCentral
49.
Maguire TM, Shering SG, Duggan CM, McDermott EW, O’Higgins NJ, Duffy MJ. High levels of cathepsin B predict poor outcome in patients with breast cancer. Int J Biol Markers. 1998;13:139–44.PubMed
50.
Wolff C, Malinowsky K, Berg D, Schragner K, Schuster T, Walch A, et al. Signalling networks associated with urokinase-type plasminogen activator (uPA) and its inhibitor PAI-1 in breast cancer tissues: new insights from protein microarray analysis. J Pathol. 2011;223:54–63.CrossRefPubMed
Metadata
Title
uPAR enhances malignant potential of triple-negative breast cancer by directly interacting with uPA and IGF1R
Authors
Michaela C. Huber
Rebecca Mall
Herbert Braselmann
Annette Feuchtinger
Sara Molatore
Katrin Lindner
Axel Walch
Eva Gross
Manfred Schmitt
Natalie Falkenberg
Michaela Aubele
Publication date
01-12-2016
Publisher
BioMed Central
Published in
BMC Cancer / Issue 1/2016
Electronic ISSN: 1471-2407
DOI
https://doi.org/10.1186/s12885-016-2663-9

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