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.

  • Original Article
  • Published:

Loss of responsiveness to IGF-I in cells with reduced cathepsin L expression levels

Oncogene volume 27, pages 4973–4985 (2008)Cite this article

Abstract

The lysosomal cysteine proteinase cathepsin L is involved in proteolytic processing of internalized proteins. In transformed cells, where it is frequently overexpressed, its intracellular localization and functions can be altered. Previously, we reported that treatment of highly metastatic, murine carcinoma H-59 cells with small molecule cysteine proteinase inhibitors altered the responsiveness of the type I insulin-like growth factor (IGF-I) receptor and consequently reduced cell invasion and metastasis. To assess more specifically the role of cathepsin L in IGF-I-induced signaling and tumorigenicity, we generated H-59 subclones with reduced cathepsin L expression levels. These clonal lines showed an altered responsiveness to IGF-I in vitro, as evidenced by (i) loss of IGF-I-induced receptor phosphorylation and Shc recruitment, (ii) reduced IGF-I (but not IGF-II)-induced cellular proliferation and migration, (iii) decreased anchorage-independent growth and (iv) reduced plasma membrane levels of IGF-IR. These changes resulted in increased apoptosis in vivo and an impaired ability of the cells to form liver metastases. The results demonstrate that cathepsin L expression levels regulate cell responsiveness to IGF-I and thereby identify a novel function for cathepsin L in the control of the tumorigenic/metastatic phenotype.

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 7
Figure 6

Similar content being viewed by others

References

  • Ahn K, Yeyeodu S, Collette J, Madden V, Arthur J, Li L et al. (2002). An alternate targeting pathway for procathepsin L in mouse fibroblasts. Traffic 3: 147–159.

    Article  CAS  Google Scholar 

  • Authier F, Kouach M, Briand G . (2005). Endosomal proteolysis of insulin-like growth factor-I at its C-terminal D-domain by cathepsin B. FEBS Lett 579: 4309–4316.

    Article  CAS  Google Scholar 

  • Authier F, Metioui M, Bell AW, Mort JS . (1999). Negative regulation of epidermal growth factor signaling by selective proteolytic mechanisms in the endosome mediated by cathepsin B. J Biol Chem 274: 33723–33731.

    Article  CAS  Google Scholar 

  • Authier F, Metioui M, Fabrega S, Kouach M, Briand G . (2002). Endosomal proteolysis of internalized insulin at the C-terminal region of the B chain by cathepsin D. J Biol Chem 277: 9437–9446.

    Article  CAS  Google Scholar 

  • Baserga R, Peruzzi F, Reiss K . (2003). The IGF-1 receptor in cancer biology. Int J Cancer 107: 873–877.

    Article  CAS  Google Scholar 

  • Benavides F, Starost MF, Flores M, Gimenez-Conti IB, Guenet JL, Conti CJ . (2002). Impaired hair follicle morphogenesis and cycling with abnormal epidermal differentiation in nackt mice, a cathepsin L-deficient mutation. Am J Pathol 161: 693–703.

    Article  CAS  Google Scholar 

  • Boike G, Lah T, Sloane BF, Rozhin J, Honn K, Guirguis R et al. (1992). A possible role for cysteine proteinase and its inhibitors in motility of malignant melanoma and other tumour cells. Melanoma Res 1: 333–340.

    Article  CAS  Google Scholar 

  • Brodt P . (1986). Characterization of two highly metastatic variants of Lewis lung carcinoma with different organ specificities. Cancer Res 46: 2442–2448.

    CAS  Google Scholar 

  • Brodt P, Fallavollita L, Khatib AM, Samani AA, Zhang D . (2001). Cooperative regulation of the invasive and metastatic phenotypes by different domains of the type I insulin-like growth factor receptor beta subunit. J Biol Chem 276: 33608–33615.

    Article  CAS  Google Scholar 

  • Brodt P, Reich R, Moroz LA, Chambers AF . (1992). Differences in the repertoires of basement membrane degrading enzymes in two carcinoma sublines with distinct patterns of site-selective metastasis. Biochim Biophys Acta 1139: 77–83.

    Article  CAS  Google Scholar 

  • Buckley DA, Loughran G, Murphy G, Fennelly C, O'Connor R . (2002). Identification of an IGF-1R kinase regulatory phosphatase using the fission yeast Schizosaccharomyces pombe and a GFP tagged IGF-1R in mammalian cells. Mol Pathol 55: 46–54.

    Article  CAS  Google Scholar 

  • Chambers AF, Colella R, Denhardt DT, Wilson SM . (1992). Increased expression of cathepsins L and B and decreased activity of their inhibitors in metastatic, ras-transformed NIH 3T3 cells. Mol Carcinog 5: 238–245.

    Article  CAS  Google Scholar 

  • Chow JC, Condorelli G, Smith RJ . (1998). Insulin-like growth factor-I receptor internalization regulates signaling via the Shc/mitogen-activated protein kinase pathway, but not the insulin receptor substrate-1 pathway. J Biol Chem 273: 4672–4680.

    Article  CAS  Google Scholar 

  • Clague MJ, Urbe S . (2001). The interface of receptor trafficking and signalling. J Cell Sci 114: 3075–3081.

    CAS  Google Scholar 

  • Collette J, Bocock JP, Ahn K, Chapman RL, Godbold G, Yeyeodu S et al. (2004). Biosynthesis and alternate targeting of the lysosomal cysteine protease cathepsin L. Int Rev Cytol 241: 1–51.

    Article  Google Scholar 

  • Denley A, Wallace JC, Cosgrove LJ, Forbes BE . (2003). The insulin receptor isoform exon 11-(IR-A) in cancer and other diseases: a review. Horm Metab Res 35: 778–785.

    Article  CAS  Google Scholar 

  • Desbuquois B, Chauvet G, Kouach M, Authier F . (2003). Cell itinerary and metabolic fate of proinsulin in rat liver: in vivo and in vitro studies. Endocrinology 144: 5308–5321.

    Article  CAS  Google Scholar 

  • Di Guglielmo GM, Baass PC, Ou WJ, Posner BI, Bergeron JJ . (1994). Compartmentalization of SHC, GRB2 and mSOS, and hyperphosphorylation of Raf-1 by EGF but not insulin in liver parenchyma. EMBO J 13: 4269–4277.

    Article  CAS  Google Scholar 

  • Gocheva V, Joyce JA . (2007). Cysteine cathepsins and the cutting edge of cancer invasion. Cell Cycle 6: 60–64.

    Article  CAS  Google Scholar 

  • Gocheva V, Zeng W, Ke D, Klimstra D, Reinheckel T, Peters C et al. (2006). Distinct roles for cysteine cathepsin genes in multistage tumorigenesis. Genes Dev 20: 543–556.

    Article  CAS  Google Scholar 

  • Gordon PV, Paxton JB, Kuemmerle JF, Fox NS . (2005). A 14-kDa cathepsin L-derived carboxyl IGFBP-2 fragment is sequestered by cultured rat ileal crypt cells. Am J Physiol Gastrointest Liver Physiol 289: G79–G87.

    Article  CAS  Google Scholar 

  • Goulet B, Sansregret L, Leduy L, Bogyo M, Weber E, Chauhan SS et al. (2007). Increased expression and activity of nuclear cathepsin L in cancer cells suggests a novel mechanism of cell transformation. Mol Cancer Res 5: 899–907.

    Article  CAS  Google Scholar 

  • Guan J, Williams C, Skinner S, Mallard E, Gluckman P . (1996). The effects of insulin-like growth factor (IGF)-1, IGF-2, and des-IGF-1 on neuronal loss after hypoxic–ischemic brain injury in adult rats: evidence for a role for IGF binding proteins. Endocrinology 137: 893–898.

    Article  CAS  Google Scholar 

  • Ishidoh K, Kominami E . (1998). Gene regulation and extracellular functions of procathepsin L. Biol Chem 379: 131–135.

    CAS  Google Scholar 

  • Ishidoh K, Kominami E . (2002). Processing and activation of lysosomal proteinases. Biol Chem 383: 1827–1831.

    Article  CAS  Google Scholar 

  • Iwata Y, Mort JS, Tateishi H, Lee ER . (1997). Macrophage cathepsin L, a factor in the erosion of subchondral bone in rheumatoid arthritis. Arthritis Rheum 40: 499–509.

    Article  CAS  Google Scholar 

  • Kirschke H, Eerola R, Hopsu-Havu VK, Bromme D, Vuorio E . (2000). Antisense RNA inhibition of cathepsin L expression reduces tumorigenicity of malignant cells. Eur J Cancer 36: 787–795.

    Article  CAS  Google Scholar 

  • Krueger S, Kellner U, Buehling F, Roessner A . (2001). Cathepsin L antisense oligonucleotides in a human osteosarcoma cell line: effects on the invasive phenotype. Cancer Gene Ther 8: 522–528.

    Article  CAS  Google Scholar 

  • Levicar N, Dewey RA, Daley E, Bates TE, Davies D, Kos J et al. (2003). Selective suppression of cathepsin L by antisense cDNA impairs human brain tumor cell invasion in vitro and promotes apoptosis. Cancer Gene Ther 10: 141–151.

    Article  CAS  Google Scholar 

  • Lin F-T, Daaka Y, Lefkowitz RJ . (1998). Beta-arrestins regulate mitogenic signaling and clathrin-mediated endocytosis of the insulin-like growth factor I receptor. J Biol Chem 273: 31640–31643.

    Article  CAS  Google Scholar 

  • Long L, Navab R, Brodt P . (1998). Regulation of the Mr 72 000 type IV collagenase by the type I insulin-like growth factor receptor. Cancer Res 58: 3243–3247.

    CAS  Google Scholar 

  • Long L, Rubin R, Baserga R, Brodt P . (1995). Loss of the metastatic phenotype in murine carcinoma cells expressing an antisense RNA to the insulin-like growth factor receptor. Cancer Res 55: 1006–1009.

    CAS  Google Scholar 

  • Meyer GE, Shelden E, Kim B, Feldman EL . (2001). Insulin-like growth factor I stimulates motility in human neuroblastoma cells. Oncogene 20: 7542–7550.

    Article  CAS  Google Scholar 

  • Mohamed MM, Sloane BF . (2006). Cysteine cathepsins: multifunctional enzymes in cancer. Nat Rev Cancer 6: 764–775.

    Article  CAS  Google Scholar 

  • Nam T, Moralez A, Clemmons D . (2002). Vitronectin binding to IGF binding protein-5 (IGFBP-5) alters IGFBP-5 modulation of IGF-I actions. Endocrinology 143: 30–36.

    Article  CAS  Google Scholar 

  • Nam TJ, Busby Jr WH, Rees C, Clemmons DR . (2000). Thrombospondin and osteopontin bind to insulin-like growth factor (IGF)-binding protein-5 leading to an alteration in IGF-I-stimulated cell growth. Endocrinology 141: 1100–1106.

    Article  CAS  Google Scholar 

  • Navab R, Chevet E, Authier F, Di Guglielmo GM, Bergeron JJ, Brodt P . (2001). Inhibition of endosomal insulin-like growth factor-I processing by cysteine proteinase inhibitors blocks receptor-mediated functions. J Biol Chem 276: 13644–13649.

    Article  CAS  Google Scholar 

  • Navab R, Mort JS, Brodt P . (1997). Inhibition of carcinoma cell invasion and liver metastases formation by the cysteine proteinase inhibitor E-64. Clin Exp Metastasis 15: 121–129.

    Article  CAS  Google Scholar 

  • Nguyen DT, Kebache S, Fazel A, Wong HN, Jenna S, Emadali A et al. (2004). Nck-dependent activation of extracellular signal-regulated kinase-1 and regulation of cell survival during endoplasmic reticulum stress. Mol Biol Cell 15: 4248–4260.

    Article  CAS  Google Scholar 

  • Nip J, Rabbani SA, Shibata HR, Brodt P . (1995). Coordinated expression of the vitronectin receptor and the urokinase-type plasminogen activator receptor in metastatic melanoma cells. J Clin Invest 95: 2096–2103.

    Article  CAS  Google Scholar 

  • Nomura T, Katunuma N . (2005). Involvement of cathepsins in the invasion, metastasis and proliferation of cancer cells. J Med Invest 52: 1–9.

    Article  Google Scholar 

  • Ou WJ, Cameron PH, Thomas DY, Bergeron JJ . (1993). Association of folding intermediates of glycoproteins with calnexin during protein maturation. Nature 364: 771–776.

    Article  CAS  Google Scholar 

  • Portnoy D, Erickson A, Kochan J, Ravetch J, Unkeless J . (1986). Cloning and characterization of a mouse cysteine proteinase. J Biol Chem 261: 14697–14703.

    CAS  Google Scholar 

  • Reinheckel T, Deussing J, Roth W, Peters C . (2001). Towards specific functions of lysosomal cysteine peptidases: phenotypes of mice deficient for cathepsin B or cathepsin L. Biol Chem 382: 735–741.

    CAS  Google Scholar 

  • Reinheckel T, Hagemann S, Dollwet-Mack S, Martinez E, Lohmuller T, Zlatkovic G et al. (2005). The lysosomal cysteine protease cathepsin L regulates keratinocyte proliferation by control of growth factor recycling. J Cell Sci 118: 3387–3395.

    Article  CAS  Google Scholar 

  • Rousselet N, Mills L, Jean D, Tellez C, Bar-Eli M, Frade R . (2004). Inhibition of tumorigenicity and metastasis of human melanoma cells by anti-cathepsin L single chain variable fragment. Cancer Res 64: 146–151.

    Article  CAS  Google Scholar 

  • Rowan AD, Mach L, Mort JS . (1992). Antibodies to rat procathepsin B recognize the active mature enzyme. Biol Chem Hoppe Seyler 373: 427–432.

    Article  CAS  Google Scholar 

  • Samani AA, Brodt P . (2001). The receptor for the type I insulin-like growth factor and its ligands regulate multiple cellular functions that impact on metastasis. Surg Oncol Clin N Am 10: 289–312, viii.

    Article  CAS  Google Scholar 

  • Samani AA, Chevet E, Fallavollita L, Galipeau J, Brodt P . (2004). Loss of tumorigenicity and metastatic potential in carcinoma cells expressing the extracellular domain of the type 1 insulin-like growth factor receptor. Cancer Res 64: 3380–3385.

    Article  CAS  Google Scholar 

  • Samani AA, Yakar S, LeRoith D, Brodt P . (2007). The role of the IGF system in cancer growth and metastasis: overview and recent insights. Endocr Rev 28: 20–47.

    Article  CAS  Google Scholar 

  • Sorkin A, Von Zastrow M . (2002). Signal transduction and endocytosis: close encounters of many kinds. Nat Rev Mol Cell Biol 3: 600–614.

    Article  CAS  Google Scholar 

  • Turk B, Turk D, Turk V . (2000). Lysosomal cysteine proteases: more than scavengers. Biochim Biophys Acta 1477: 98–111.

    Article  CAS  Google Scholar 

  • Wang N, Thuraisingam T, Fallavollita L, Ding A, Radzioch D, Brodt P . (2006). The secretory leukocyte protease inhibitor is a type 1 insulin-like growth factor receptor-regulated protein that protects against liver metastasis by attenuating the host proinflammatory response. Cancer Res 66: 3062–3070.

    Article  CAS  Google Scholar 

  • Wexler H . (1966). Accurate identification of experimental pulmonary metastases. J Natl Cancer Inst 36: 641–645.

    Article  CAS  Google Scholar 

  • Yang M, Zhang Y, Pan J, Sun J, Liu J, Libby P et al. (2007). Cathepsin L activity controls adipogenesis and glucose tolerance. Nat Cell Biol 9: 970–977.

    Article  CAS  Google Scholar 

  • Yang Z, Cox JL . (2007). Cathepsin L increases invasion and migration of B16 melanoma. Cancer Cell Int 7: 8.

    Article  Google Scholar 

  • Zhang D, Bar-Eli M, Meloche S, Brodt P . (2004). Dual Regulation of MMP-2 Expression by the Type 1 Insulin-like Growth Factor Receptor: the PI 3-kinase/Akt and Raf/MAP kinase pathways transmit opposing signals. J Biol Chem 279: 19683–19690.

    Article  CAS  Google Scholar 

  • Zhang D, Brodt P . (2003). Type 1 insulin-like growth factor regulates MT1-MMP synthesis and tumor invasion via PI 3-kinase/Akt signaling. Oncogene 22: 974–982.

    Article  CAS  Google Scholar 

  • Zwad O, Kubler B, Roth W, Scharf JG, Saftig P, Peters C et al. (2002). Decreased intracellular degradation of insulin-like growth factor binding protein-3 in cathepsin L-deficient fibroblasts. FEBS Lett 510: 211–215.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This work was supported initially by Grant MT-10505 from the Canadian Institute for Health Research and subsequently by a Terry Fox Frontiers Initiative Grant from the National Cancer Institute of Canada (to PB) and by Grant MCB-96141139 from the National Science Foundation (to AE). We thank Dr Edmund Ziomek (Biotechnology Research Institute, Montreal, QC, Canada) for gifts of CathC and CathS cDNA, and Drs Amir Samani and Donglei Zhang (formerly of the Division of Surgical Research, McGill University) for their help with the study.

Author information

Author notes

  1. R Navab

    Present address: 11Current address: Lung Cancer Translational Laboratory, Princess Margaret Hospital, University Health Network, Toronto, Ontario, Canada,

  2. C Pedraza

    Present address: 12Current address: Department of Dentistry, McGill University, Montreal, Quebec, Canada.,

  3. S Jenna

    Present address: 13Current address: Department of Chemistry-Biochemistry, Université du Québec à Montréal, Montréal, Canada.,

Authors and Affiliations

  1. Department of Surgery, McGill University, Montreal, Quebec, Canada

    R Navab, C Pedraza, L Fallavollita, N Wang, E Chevet, Z You, J S Mort & P Brodt

  2. Department of Medicine, McGill University, Montreal, Quebec, Canada

    E Chevet & P Brodt

  3. The Organelle Signaling Laboratory, McGill University Health Center—Royal Victoria Hospital, McGill University, Montreal, Quebec, Canada

    E Chevet, P Auguste & S Jenna

  4. Team Avenir, INSERM U889, Université Bordeaux 2, Bordeaux, France

    E Chevet & P Auguste

  5. INSERM U889, University Bordeaux1, Bordeaux, France

    P Auguste

  6. INSERM E0113, Université Bordeaux 1, Bordeaux, France

    A Bikfalvi

  7. Lung Biology Research, Hospital for Sick Children, University of Toronto, Toronto, Ontario, Canada

    J Hu

  8. Department of Biochemistry, University College Cork, Cork, Ireland

    R O'Connor

  9. Department of Biochemistry and Biophysics, University of North Carolina Chapel Hill, North Carolina, NC, USA

    A Erickson

  10. The Shriners Hospital, McGill University, Montreal, Quebec, Canada

    J S Mort

Authors
  1. R Navab
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. C Pedraza
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. L Fallavollita
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. N Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. E Chevet
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. P Auguste
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. S Jenna
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Z You
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. A Bikfalvi
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. J Hu
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. R O'Connor
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. A Erickson
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. J S Mort
    View author publications

    You can also search for this author in PubMed Google Scholar

  14. P Brodt
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to P Brodt.

Additional information

Supplementary Information accompanies the paper on the Oncogene website (http://www.nature.com/onc).

Supplementary information

Rights and permissions

Reprints and permissions

About this article

Cite this article

Navab, R., Pedraza, C., Fallavollita, L. et al. Loss of responsiveness to IGF-I in cells with reduced cathepsin L expression levels. Oncogene 27, 4973–4985 (2008). https://doi.org/10.1038/onc.2008.144

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/onc.2008.144

Keywords

This article is cited by

Search

Advanced search

Quick links