The online version of this article (https://doi.org/10.1186/s13058-018-0936-8) contains supplementary material, which is available to authorized users.
The Fos-related antigen 1 (FRA-1) transcription factor promotes tumor cell growth, invasion and metastasis. Phosphorylation of FRA-1 increases protein stability and function. We identify a novel signaling axis that leads to increased phosphorylation of FRA-1, increased extracellular matrix (ECM)-induced breast cancer cell invasion and is prognostic of poor outcome in patients with breast cancer.
While characterizing five breast cancer cell lines derived from primary human breast tumors, we identified BRC-31 as a novel basal-like cell model that expresses elevated FRA-1 levels. We interrogated the functional contribution of FRA-1 and an upstream signaling axis in breast cancer cell invasion. We extended this analysis to determine the prognostic significance of this signaling axis in samples derived from patients with breast cancer.
BRC-31 cells display elevated focal adhesion kinase (FAK), SRC and extracellular signal-regulated (ERK2) phosphorylation relative to luminal breast cancer models. Inhibition of this signaling axis, with pharmacological inhibitors, reduces the phosphorylation and stabilization of FRA-1. Elevated integrin αVβ3 and uPAR expression in these cells suggested that integrin receptors might activate this FAK-SRC-ERK2 signaling. Transient knockdown of urokinase/plasminogen activator urokinase receptor (uPAR) in basal-like breast cancer cells grown on vitronectin reduces FRA-1 phosphorylation and stabilization; and uPAR and FRA-1 are required for vitronectin-induced cell invasion. In clinical samples, a molecular component signature consisting of vitronectin-uPAR-uPA-FRA-1 predicts poor overall survival in patients with breast cancer and correlates with an FRA-1 transcriptional signature.
We have identified a novel signaling axis that leads to phosphorylation and enhanced activity of FRA-1, a transcription factor that is emerging as an important modulator of breast cancer progression and metastasis.
Additional file 1: Oligonucleotides utilized in this manuscript. (XLSX 47 kb)13058_2018_936_MOESM1_ESM.xlsx
Additional file 2: Document 1: Supplemental Figure legends and Methods. (DOCX 86 kb)13058_2018_936_MOESM2_ESM.docx
Additional file 3: Figure S1. Gene Expression of mapk1, mapk3 and fosl1 in human Breast Cancer Cell lines. (PPTX 1395 kb)13058_2018_936_MOESM3_ESM.pptx
Additional file 4: Figure S2. EGFR inhibition is not sufficient to decrease phosphorylation on FRA-1. (PPTX 4508 kb)13058_2018_936_MOESM4_ESM.pptx
Additional file 5: Figure S3. FRA-1 phosphorylation occurs prior to cell spreading. (PPTX 4024 kb)13058_2018_936_MOESM5_ESM.pptx
Additional file 6: Figure S4. Gene Expression of plaur in human Breast Cancer Cell lines. (PPTX 1129 kb)13058_2018_936_MOESM6_ESM.pptx
Additional file 7: Figure S5. Knockdown of plaur or fosl1 does not affect cell proliferation. (PPTX 515 kb)13058_2018_936_MOESM7_ESM.pptx
Additional file 8: Figure S6. Basal-like breast cancer cell lines and patient-derived xenografts (PDXs) that possess elevated FRA-1 phosphorylation display high uPAR and uPA expression. (PPTX 1129 kb)13058_2018_936_MOESM8_ESM.pptx
Dhillon AS, Tulchinsky E. FRA-1 as a driver of tumour heterogeneity: a nexus between oncogenes and embryonic signalling pathways in cancer. Oncogene. 2015; 34(34):4421-8.
Maurus K, Hufnagel A, Geiger F, Graf S, Berking C, Heinemann A, Paschen A, Kneitz S, Stigloher C, Geissinger E, et al. The AP-1 transcription factor FOSL1 causes melanocyte reprogramming and transformation. Oncogene. 2017.
Oliveira-Ferrer L, Kurschner M, Labitzky V, Wicklein D, Muller V, Luers G, Schumacher U, Milde-Langosch K, Schroder C. Prognostic impact of transcription factor Fra-1 in ER-positive breast cancer: contribution to a metastatic phenotype through modulation of tumor cell adhesive properties. J Cancer Res Clin Oncol. 2015;141(10):1715-26.
Stinson S, Lackner MR, Adai AT, Yu N, Kim HJ, O'Brien C, Spoerke J, Jhunjhunwala S, Boyd Z, Januario T, et al. TRPS1 targeting by miR-221/222 promotes the epithelial-to-mesenchymal transition in breast cancer. Sci Signal. 2011;4(177):ra41.
Leconet W, Chentouf M, du Manoir S, Chevalier C, Sirvent A, Ait-Arsa I, Busson M, Jarlier M, Radosevic-Robin N, Theillet C et al. Therapeutic activity of anti-AXL antibody against triple-negative breast cancer patient-derived xenografts and metastasis. Clin Cancer Res. 2017;23(11):2806–16. CrossRefPubMed
Chen RH, Juo PC, Curran T, Blenis J. Phosphorylation of c-Fos at the C-terminus enhances its transforming activity. Oncogene. 1996;12(7):1493–502. PubMed
Puiffe ML, Le Page C, Filali-Mouhim A, Zietarska M, Ouellet V, Tonin PN, Chevrette M, Provencher DM, Mes-Masson AM. Characterization of ovarian cancer ascites on cell invasion, proliferation, spheroid formation, and gene expression in an in vitro model of epithelial ovarian cancer. Neoplasia. 2007;9(10):820–9. CrossRefPubMedPubMedCentral
Xue W, Mizukami I, Todd 3rd RF, Petty HR. Urokinase-type plasminogen activator receptors associate with beta1 and beta3 integrins of fibrosarcoma cells: dependence on extracellular matrix components. Cancer Res. 1997;57(9):1682–9. PubMed
Zhang X, Claerhout S, Prat A, Dobrolecki LE, Petrovic I, Lai Q, Landis MD, Wiechmann L, Schiff R, Giuliano M, et al. A renewable tissue resource of phenotypically stable, biologically and ethnically diverse, patient-derived human breast cancer xenograft models. Cancer Res. 2013;73(15):4885–97. CrossRefPubMedPubMedCentral
Pirazzoli V, Ferraris GMS, Sidenius N. Direct evidence of the importance of vitronectin and its interaction with the urokinase receptor in tumor growth. Blood. 2013;121(12):2316-23.
Kadowaki M, Sangai T, Nagashima T, Sakakibara M, Yoshitomi H, Takano S, Sogawa K, Umemura H, Fushimi K, Nakatani Y, et al. Identification of vitronectin as a novel serum marker for early breast cancer detection using a new proteomic approach. J Cancer Res Clin Oncol. 2011;137(7):1105–15. CrossRefPubMed
von Thun A, Preisinger C, Rath O, Schwarz JP, Ward C, Monsefi N, Rodríguez J, Garcia-Munoz A, Birtwistle M, Bienvenut W, et al. Extracellular signal-regulated kinase regulates rhoa activation and tumor cell plasticity by inhibiting guanine exchange factor H1 activity. Mol Cell Biol. 2013;33(22):4526–37. CrossRef
Ellis V, Behrendt N, Dano K. Plasminogen activation by receptor-bound urokinase. A kinetic study with both cell-associated and isolated receptor. J Biol Chem. 1991;266(19):12752–8. PubMed
Riisbro R, Christensen IJ, Piironen T, Greenall M, Larsen B, Stephens RW, Han C, Hoyer-Hansen G, Smith K, Brunner N, et al. Prognostic significance of soluble urokinase plasminogen activator receptor in serum and cytosol of tumor tissue from patients with primary breast cancer. Clin Cancer Res. 2002;8(5):1132–41. PubMed
Ragone C, Minopoli M, Ingangi V, Botti G, Fratangelo F, Pessi A, Stoppelli MP, Ascierto PA, Ciliberto G, Motti ML, et al. Targeting the cross-talk between urokinase receptor and formyl peptide receptor type 1 to prevent invasion and trans-endothelial migration of melanoma cells. J Exp Clin Cancer Res. 2017;36(1):180. CrossRefPubMedPubMedCentral
- Integrin-uPAR signaling leads to FRA-1 phosphorylation and enhanced breast cancer invasion
Matthew G. Annis
Jonathan P. Rennhack
Eran R. Andrechek
Peter M. Siegel
- BioMed Central
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