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 Paper
  • Published:

A mutated cathepsin-D devoid of its catalytic activity stimulates the growth of cancer cells

Oncogene volume 20, pages 6920–6929 (2001)Cite this article

Abstract

Cathepsin-D, a lysosomal aspartyl proteinase, is highly secreted by breast cancer cells and its over-expression by transfection stimulates cancer cell proliferation. The mechanism by which this protease affects proliferation remains, however, unknown. In order to determine whether proteolytic activity is necessary, we abolished its enzymatic activity using site-directed mutagenesis followed by stable transfection in 3Y1-Ad12 cancer cells. Substitution of the aspartic acid residue 231 by an asparagine residue in its catalytic site abrogated the cathepsin-D proteolytic activity but did not affect its expression level, processing or secretion. However, like wild-type cathepsin-D, this mutated catalytically-inactive cathepsin-D retained its capacity to stimulate proliferation of cells embedded in Matrigel or collagen I matrices, colony formation in soft agar and tumor growth in athymic nude mice. Addition on the mock-transfected cells, of either conditioned media containing the wild-type or the mutated pro-cathepsin-D, or of the purified mutated pro-cathepsin-D, partially mimicked the mitogenic activity of the transfected cathepsin-D, indicating a role of the secreted pro-enzyme. Moreover, addition of two anti-cathepsin-D antibodies on the cathepsin-D transfected cells inhibited their proliferation, suggesting an action of the secreted pro-cathepsin-D via an autocrine loop. A synthetic peptide containing the 27-44 residue moiety of the cathepsin-D pro-fragment was, however, not mitogenic suggesting that a receptor for the pro-fragment was not involved. Furthermore, the cathepsin-D mitogenicity was not blocked by inhibiting the interaction of pro-cathepsin-D with the mannose-6-phosphate receptors. Our results altogether demonstrate that a mutated cathepsin-D devoid of catalytic activity is still mitogenic and suggest that it is acting extra-cellularly by triggering directly or indirectly a yet unidentified cell surface receptor.

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
Figure 8
Figure 9

Similar content being viewed by others

References

  • Bissell MJ, Weaver VM, Lelièvre SA, Wang F, Petersen EW, Schmeichel KL . 1999 Cancer Res. 59: 1757s–1764s

  • Briozzo P, Badet J, Capony F, Pieri I, Montcourrier P, Barritault D, Rochefort H . 1991 Exp. Cell Res. 194: 252–259

  • Capony F, Morisset M, Barrett AJ, Capony F, Broquet P, Vignon F, Chambon M, Louisot P, Rochefort H . 1987 J. Cell. Biol. 104: 253–262

  • Capony F, Rougeot C, Montcourier P, Cavaillès V, Salazar G, Rochefort H . 1989 Cancer Res. 49: 3904–3909

  • Carney DH, Cunningham DD . 1978 Cell 15: 1341–1349

  • Chambers AF, Matrisian LM . 1997 J. Natl. Cancer I. 89: 1260–1270

  • De Clerck YA, Imren S . 1994 Eur. J. Cancer 30A: 2170–2180

  • De Leon DD, Terry C, Asmerom Y, Nissley P . 1996 Endocrinology 137: 1851–1859

  • Edwards DR, Murphy G . 1998 Nature 394: 527–528

  • Emerman JT, Enani J, Pitelka DR, Nandi S . 1977 Proc. Natl. Acad. Sci. USA 74: 4466–4470

  • Fang W, Hartmann N, Chow D, Riegel AT, Wellstein A . 1992 J. Biol. Chem. 267: 25889–25897

  • Faust PL, Kornfeld S, Chirgwin JM . 1985 Proc. Natl. Acad. Sci. USA 82: 4910–4914

  • Ferrandina G, Scambia G, Bardelli F, Panici B, Mancuso S, Messori A . 1997 Br. J. Cancer 76: 661–666

  • Foekens JA, Look MP, Bolt-de Vries J, Meijer-van Gelder ME, van Putten WLJ, Klijn JGM . 1999 Br. J. Cancer 79: 300–307

  • Freiss G, Vignon F, Rochefort H . 1988 Cancer Res. 48: 3709–3715

  • Fusek M, Vetvicka VJ . 1994 Biochem. J. 303: 775–780

  • Garcia M, Capony F, Derocq D, Simon D, Pau B, Rochefort H . 1985 Cancer Res. 45: 709–716

  • Garcia M, Derocq D, Pujol P, Rochefort H . 1990 Oncogene 5: 1809–1814

  • Kane SE, Gottesman MM . 1990 Semin. Cancer Biol. 1: 127–136

  • Kohl NE, Emini EA, Schleif WA, Davis LJ, Heimbach JC, Dixon RA, Scolnick EM, Sigal IS . 1988 Proc. Natl. Acad. Sci. USA 85: 4686–4690

  • Laurent-Matha V, Farnoud MR, Lucas A, Rougeot C, Garcia M, Rochefort H . 1998 J. Cell Sci. 111: 2539–2549

  • Liaudet E, Derocq D, Rochefort H, Garcia M . 1995 Cell Growth Differ. 6: 1045–1052

  • Lin XL, Wong RN, Tang J . 1989 J. Biol. Chem. 264: 4482–4489

  • Liotta LA, Rao CN, Wewer UM . 1986 Ann. Rev. Biochem. 55: 1037–1057

  • Mathieu M, Rochefort H, Barenton B, Prebois C, Vignon F . 1990 Mol. Endocrinol. 4: 1327–1335

  • Metcalf P, Fusek M . 1993 EMBO J. 12: 1293–1302

  • Montcourrier P, Mangeat PH, Valembois C, Salazar G, Sahuquet A, Duperray C, Rochefort H . 1994 J. Cell. Sci. 107: 2381–2391

  • Montcourrier P, Silver IA, Farnoud R, Bird I, Rochefort H . 1997 Clin. Exp. Metast. 15: 382–392

  • Rifkin DB . 1997 Fibrinol. Proteol. 11: 3–9

  • Rochefort H, Capony F, Garcia M, Cavaillès V, Freiss G, Chambon M, Morisset M, Vignon F . 1987 J. Cell. Biochem. 35: 17–29

  • Rochefort H . 1992 Eur. J. Cancer 28A: 1780–1783

  • Rochefort H, Liaudet-Coopman E . 1999 APMIS 107: 86–95

  • Seelmeier S, Schmidt H, Turk V, Von Der Helm K . 1988 Proc. Natl. Acad. Sci. USA 85: 6612–6616

  • Sloane BF, Honn KV . 1984 Cancer Metastasis Rev. 3: 249–263

  • Thompson EW, Paik S, Brünner N, Sommers CL, Zugmaier G, Clarke R, Shima TB, Torri J, Donahue S, Lippman ME, Martin GR, Dickson RB . 1992 J. Cell. Physiol. 150: 534–544

  • Veith FO, Capony F, Garcia M, Chantelard J, Pujol H, Veith F, Zajdela A, Rochefort H . 1983 Cancer Res. 43: 1861–1868

  • Vetvicka V, Vetvickova J, Fusek M . 1999 Breast Cancer Res. Treat. 57: 261–269

  • Vignon F, Capony F, Chambon M, Freiss G, Garcia M, Rochefort H . 1986 Endocrinology 118: 1537–1545

  • Vignon F, Rochefort H . 1992 Breast Cancer Res. Treat. 22: 47–57

  • Wittlin S, Rosel J, Hofmann F, Stover DR . 1999 Eur. J. Biochem. 265: 384–393

Download references

Acknowledgements

We thank Dr F Vignon for helpful discussions and suggestions, Dr P Roger for pathology advice, V Gaubiac, M Gleizes and A Lucas for technical assistance, JY Cance for photographs and drawing of figures, and N Kerdjadj and E Moreno for secretarial assistance. This work was supported by the University of Montpellier I, the ‘Institut National de la Santé et de la Recherche Médicale,’ the ‘Association pour la Recherche sur le Cancer,’ the ‘Institut de Recherches Servier’ and the ‘Ligue Nationale contre le Cancer, Comité de l'Hérault’.

Author information

Authors and Affiliations

  1. INSERM U540 ‘Endocrinologie Moléculaire et Cellulaire des Cancers’, Université de Montpellier 1, 60 rue de Navacelles, 34090, Montpellier, France

    Murielle Glondu, Valérie Laurent-Matha, Marcel Garcia, Henri Rochefort & Emmanuelle Liaudet-Coopman

  2. CNRS UMR 5539, Université de Montpellier 2, Place Eugène Bataillon, Montpellier, 34095, France

    Peter Coopman

Authors
  1. Murielle Glondu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Peter Coopman
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Valérie Laurent-Matha
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Marcel Garcia
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Henri Rochefort
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Emmanuelle Liaudet-Coopman
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Henri Rochefort.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Glondu, M., Coopman, P., Laurent-Matha, V. et al. A mutated cathepsin-D devoid of its catalytic activity stimulates the growth of cancer cells. Oncogene 20, 6920–6929 (2001). https://doi.org/10.1038/sj.onc.1204843

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.onc.1204843

Keywords

This article is cited by

Search

Advanced search

Quick links