Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Research Article
  • Published:

Effects of blocking urokinase receptor signaling by antisense oligonucleotides in a mouse model of experimental prostate cancer bone metastases

Gene Therapy volume 12, pages 702–714 (2005)Cite this article

Abstract

An important factor implicated in tumor cell predisposition for invasion and metastasis is the malignancy-related upregulation of urokinase plasminogen activator receptor (uPAR). uPAR signals by activating different tyrosine kinases in different cells. We examined the effects of inhibiting uPAR signaling by inhibition of uPAR expression with antisense oligonucleotides (aODNs) in PC3 human prostate cancer cells and evaluated aODN effect in a mouse model of prostate cancer bone metastasis. Following uPAR aODN treatment, PC3 cells exhibited a strong decrease in uPAR expression, evaluated by flow cytometry and by polymerase chain reaction, and of FAK/JNK/Jun phosphorylation. The synthesis of cyclins A, B, D1 and D3 was inhibited, as shown by Western blotting, flow cytometry and polymerase chain reaction, and PC3 cells accumulated in the G2 phase of the cell cycle. PC3 cells' adhesion was unaffected, while proliferation and invasion of Matrigel were impaired. A total of 60 mice were subjected to intracardiac injection of PC3 cells and were randomly assigned to three groups: aODN (treated with 0.5 mg intraperitoneum/mouse/day), dODN (treated with the same amounts of a degenerated ODN) and control (injected with a saline solution). At 28 days after heart injection, mice were subjected to a digital scan of total body radiography, which revealed 80% reduction in mice affected by bone metastasis. The use of uPAR aODNs produced a substantial prophylactic effect against prostate cancer bone metastasis, which has to be ascribed to downregulation of uPAR expression.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7

Similar content being viewed by others

References

  1. Ellis WJ et al. Detection and isolation of prostate cancer cells from peripheral blood and bone marrow. Urology 2003; 61: 277–281.

    Article  Google Scholar 

  2. Andreasen PA, Egelund R, Petersen HH . The plasminogen activation system in tumor growth, invasion, and metastasis. Cell Mol Life Sci 2000; 57: 25–40.

    Article  CAS  Google Scholar 

  3. Mignatti P, Rifkin DB . Nonenzymatic interactions between proteinases and the cell surface: novel roles in normal and malignant cell physiology. Adv Cancer Res 2000; 78: 103–157.

    Article  CAS  Google Scholar 

  4. Del Rosso M et al. Multiple pathways of cell invasion are regulated by multiple families of serine proteases. Clin Exp Metast 2002; 19: 193–207.

    Article  CAS  Google Scholar 

  5. Aguirre Ghiso JA, Kovalski K, Ossowski L . Tumor dormancy induced by down-regulation of urokinase receptor in human carcinoma involves integrin and MAPK signalling. J Cell Biol 1999; 147: 89–104.

    Article  CAS  Google Scholar 

  6. Aguirre Ghiso JA . Inhibition of FAK signalling activated by urokinase receptor induces dormancy in human carcinoma cells in vivo. Oncogene 2002; 21: 2513–2524.

    Article  Google Scholar 

  7. Aguirre Ghiso JA et al. Urokinase receptor and fibronectin regulate the ERK (MAPK) to p38 (MAPK) activity ratios that determine carcinoma cell proliferation or dormancy in vivo. Mol Biol Cell 2001; 12: 863–879.

    Article  CAS  Google Scholar 

  8. Quattrone A et al. Reversion of the invasive phenotype of transformed human fibroblasts by anti-messenger oligonucleotide inhibition of urokinase receptor gene expression. Cancer Res 1995; 55: 90–95.

    CAS  Google Scholar 

  9. Fibbi G et al. Functions of the fibrinolytic system in human Ito cells and its control by basic fibroblast and platelet-derived growth factor. Hepatology 1999; 29: 868–878.

    Article  CAS  Google Scholar 

  10. Aguirre-Ghiso JA, Estrada Y, Lu D, Ossowski L . ERKMAPK activity as a determinant of tumor growth and dormancy; regulation by p38SAPK. Cancer Res 2003; 63: 1684–1695.

    CAS  Google Scholar 

  11. Goltzman D . Mechanisms of the development of osteoblastic metastases. Cancer 1997; 80 (Suppl. 8): 1581–1587.

    Article  CAS  Google Scholar 

  12. Rabbani SA, Xing RH . Role of urokinase (uPA) and its receptor (uPAR) in invasion and metastasis of hormone-dependent malignancies. Int J Oncol 1998; 12: 911–920.

    CAS  PubMed  Google Scholar 

  13. Festuccia C et al. Plasminogen activator system modulates invasive capacity and proliferation in prostatic tumor cells. Clin Exp Metast 1998; 16: 513–528.

    Article  CAS  Google Scholar 

  14. Gavrilov D et al. Expression of urokinase plasminogen activator and receptor in conjunction with the ets family and AP-1 complex transcription factors in high grade prostate cancers. Eur J Cancer 2001; 37: 1033–1040.

    Article  CAS  Google Scholar 

  15. Miyake H et al. Elevation of serum levels of urokinase-type plasminogen activator and its receptor is associated with disease progression and prognosis in patients with prostate cancer. Prostate 1999; 39: 123–129.

    Article  CAS  Google Scholar 

  16. Magdolen V et al. Inhibition of the tumor-associated urokinase-type plasminogen activation system: effects of high-level synthesis of soluble urokinase receptor in ovarian and breast cancer cells in vitro and in vivo. Recent Results Cancer Res 2003; 162: 43–63.

    Article  CAS  Google Scholar 

  17. Kobayashi H et al. Bikunin plus paclitaxel markedly reduces tumor burden and ascites in mouse model of ovarian cancer. Int J Cancer 2004; 110: 134–139.

    Article  CAS  Google Scholar 

  18. D'Alessio S et al. Antisense oligodeoxynucleotides for urokinase-plasminogen activator receptor have anti-invasive and anti-proliferative effects in vitro and inhibit spontaneous metastases of human melanoma in mice. Int J Cancer 2004; 110: 125–133.

    Article  CAS  Google Scholar 

  19. Yang YM et al. C-Jun NH(2)-terminal kinase mediates proliferation and tumor growth of human prostate carcinoma. Clin Cancer Res 2003; 9: 391–401.

    CAS  Google Scholar 

  20. Shaulian E, Karin M . AP-1 in cell proliferation and survival. Oncogene 2001; 20: 2390–2400.

    Article  CAS  Google Scholar 

  21. Schwabe RF et al. c-Jun-N-terminal kinase drives cyclin D1 expression and proliferation during liver regeneration. Hepatology 2003; 37: 824–832.

    Article  CAS  Google Scholar 

  22. Bakiri L, Lallemand D, Bossy-Wetzel E, Yaniv M . Cell cycle-dependent variations in c-Jun and JunB phosphorylation: a role in the control of cyclin D1 expression. EMBO J 2000; 19: 2056–2068.

    Article  CAS  Google Scholar 

  23. Oktay M et al. Integrin-mediated activation of focal adhesion kinase is required for signaling to Jun NH2-terminal kinase and progression through the G1 phase of the cell cycle. J Cell Biol 1999; 145: 1461–1469.

    Article  CAS  Google Scholar 

  24. Bohuslav J et al. Urokinase plasminogen activator receptor, beta 2-integrins, and Src-kinases within a single receptor complex of human monocytes. J Exp Med 1995; 181: 1381–1390.

    Article  CAS  Google Scholar 

  25. Xue W, Kindzelskii AL, Todd III RF, Petty HR . Physical association of complement receptor type 3 and urokinase-type plasminogen activator receptor in neutrophil membranes. J Immunol 1994; 152: 4630–4640.

    CAS  PubMed  Google Scholar 

  26. Wei Y et al. Regulation of integrin function by the urokinase receptor. Science 1996; 273: 1551–1555.

    Article  CAS  Google Scholar 

  27. Montuori N et al. The cleavage of the urokinase receptor regulates its multiple functions. J Biol Chem 2002; 277: 46932–46939.

    Article  CAS  Google Scholar 

  28. Koshelnick Y et al. Urokinase receptor is associated with the components of the JAK1/STAT1 signaling pathway and leads to activation of this pathway upon receptor clustering in the human kidney epithelial tumor cell line TCL-598. J Biol Chem 1997; 272: 28563–28567.

    Article  CAS  Google Scholar 

  29. Behrendt N et al. A urokinase receptor-associated protein with specific collagen binding properties. J Biol Chem 2000; 275: 1993–2002.

    Article  CAS  Google Scholar 

  30. Ossowski L, Aguirre-Ghiso JA . Urokinase receptor and integrin partnership: coordination of signaling for cell adhesion, migration and growth. Curr Opin Cell Biol 2000; 12: 613–620.

    Article  CAS  Google Scholar 

  31. Blasi F, Carmeliet P . uPAR: a versatile signalling orchestrator. Nat Rev Mol Cell Biol 2002; 3: 932–943.

    Article  CAS  Google Scholar 

  32. Lucia MS et al. Workgroup I: rodent models of prostate cancer. Prostate 1998; 36: 49–55.

    Article  CAS  Google Scholar 

  33. Soos G et al. Heterotypic growth of human prostate carcinoma in the femurs of nude mice: an osseous metastatic model. Int J Cancer 1996; 66: 280–281.

    Article  CAS  Google Scholar 

  34. Shevrin DH, Kukreja SC, Ghosh L, Lad TE . Development of skeletal metastasis by human prostate cancer in athymic nude mice. Clin Exp Metast 1988; 6: 401–409.

    Article  CAS  Google Scholar 

  35. Wu TT et al. Establishing human prostate cancer cell xenografts in bone: induction of osteoblastic reaction by prostate-specific antigen-producing tumors in athymic and SCID/bg mice using LNCaP and lineage-derived metastatic sublines. Int J Cancer 1998; 77: 887–894.

    Article  CAS  Google Scholar 

  36. Arguello F, Baggs RB, Frantz CN . A murine model of experimental metastasis to bone and bone marrow. Cancer Res 1988; 48: 6876–6881.

    CAS  PubMed  Google Scholar 

  37. Wetterwald A et al. Optical imaging of cancer metastasis to bone marrow: a mouse model of minimal residual disease. Am J Pathol 2002; 160: 1143–1153.

    Article  Google Scholar 

  38. Corey E et al. Establishment and characterization of osseous prostate cancer models: intra-tibial injection of human prostate cancer cells. Prostate 2002; 52: 20–33.

    Article  Google Scholar 

  39. Shetty S, Kumar A, Johnson AR, Idell S . Regulation of mesothelial cell mitogenesis by antisense oligonucleotides for the urokinase receptor. Antisense Res Dev 1995; 5: 307–314.

    Article  CAS  Google Scholar 

  40. Kook YH, Adamski J, Zelent A, Ossowski L . The effect of antisense inhibition of urokinase receptor in human squamous cell carcinoma on malignancy. EMBO J 1994; 13: 3983–3991.

    Article  CAS  Google Scholar 

  41. Go Y et al. Inhibition of in vivo tumorigenicity and invasiveness of a human glioblastoma cell line transfected with antisense uPAR vectors. Clin Exp Metasts 1997; 15: 440–446.

    Article  CAS  Google Scholar 

  42. Fibbi G et al. Urokinase-dependent angiogenesis in vitro and diacylglycerol production are blocked by antisense oligonucleotides against the urokinase receptor. Lab Invest 1998; 78: 1109–1119.

    CAS  Google Scholar 

  43. Annunziato F et al. MDC and ELC attract human thymocytes in different stages of development and are produced by distinct subsets of medullary epithelial cells: possible implications for negative selection. J Immunol 2000; 165: 238–246.

    Article  CAS  Google Scholar 

  44. Romagnani P et al. Interferon-inducible protein 10, monokine induced by interferon γ, and interpheron-inducible T-cell α chemoattractant are produced by thymic epithelial cells and attract T-cell receptor (TCR) αβ1 CD8+ single-positive T cells, TCRγδ1 T cells, and natural killer-type cells in human thymus. Blood 2001; 97: 601–607.

    Article  CAS  Google Scholar 

  45. Romagnani P et al. Cell cycle-dependent expression of CXC chemokine receptor 3 by endothelial cells mediates angiostatic activity. J Clin Invest 2001; 107: 23–33.

    Article  Google Scholar 

  46. Arguello F, Baggs RB, Frantz CN . A murine model of experimental metastasis to bone and bone marrow. Cancer Res 1988; 48: 6876–6881.

    CAS  PubMed  Google Scholar 

  47. Agrawal S, Temsamani J, Tang JY . Pharmacokinetics, biodistribution, and stability of oligodeoxynucleotide phosphorothioates in mice. Proc Natl Acad Sci USA 1991; 88: 7595–7599.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This study was supported by grants from University of Florence, and Ente Cassa di Risparmio of Florence. We acknowledge the skilful technical assistance of Mr Marco Cutri of the Department of Experimental Pathology and Oncology (University of Florence) with computer programs.

Author information

Authors and Affiliations

  1. Department of Experimental Pathology and Oncology, University of Florence, Italy

    F Margheri, S D'Alessio, S Serratí, M Pucci, G Fibbi & M Del Rosso

  2. Department of Internal Medicine, University of Florence, Florence, Italy

    F Annunziato, L Cosmi, F Liotta & R Angeli

  3. Department of Experimental Medicine, University of L'Aquila Medical School, Italy

    A Angelucci, G L Gravina, N Rucci, M Bologna & A Teti

  4. Isis Pharmaceuticals, Carlsbad Research Center, Carlsbad, CA, USA

    B Monia

Authors
  1. F Margheri
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. S D'Alessio
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. S Serratí
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. M Pucci
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. F Annunziato
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. L Cosmi
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. F Liotta
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. R Angeli
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. A Angelucci
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. G L Gravina
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. N Rucci
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. M Bologna
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. A Teti
    View author publications

    You can also search for this author in PubMed Google Scholar

  14. B Monia
    View author publications

    You can also search for this author in PubMed Google Scholar

  15. G Fibbi
    View author publications

    You can also search for this author in PubMed Google Scholar

  16. M Del Rosso
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Margheri, F., D'Alessio, S., Serratí, S. et al. Effects of blocking urokinase receptor signaling by antisense oligonucleotides in a mouse model of experimental prostate cancer bone metastases. Gene Ther 12, 702–714 (2005). https://doi.org/10.1038/sj.gt.3302456

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.gt.3302456

Keywords

This article is cited by

Search

Advanced search

Quick links