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Cathepsin D Overexpressed by Cancer Cells Can Enhance Apoptosis-dependent Chemo-sensitivity Independently of Its Catalytic Activity

  • Melanie Beaujouin
  • Emmanuelle Liaudet-Coopman
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 617)

Summary

The aspartic protease cathepsin D (CD) is a key mediator of inducedapoptosis and its proteolytic activity has been generally involved in this event. During apoptosis, CD is translocated to the cytosol. Since CD is one of the lysosomal enzymes that requires a more acidic pH to be proteolytically-active relative to the cysteine lysosomal enzymes such as cathepsin-B and cathepsin-L, it is therefore open to question whether cytosolic CD might be able to cleave substrate(s) implicated in the apoptotic cascade. Here, we have investigated the role of (wild-type) wt CD and its proteolytically inactive counterpart overexpressed by 3Y1-Adl2 cancer cells during chemotherapeutic-induced cytotoxicity and apoptosis, as well as the relevance of CD catalytic function. We demonstrate that wt or mutated catalytically inactive CD strongly enhances chemo-sensitivity and apoptotic response to etoposide. Both wt and mutated inactive CD are translocated to the cytosol, increasing the release of cytochrome c, the activation of caspases-9 and caspases-3 and the induction of a caspase-dependent apoptosis. In addition, pretreatment of cells with the aspartic protease inhibitor, pepstatin A, does not prevent apoptosis. Interestingly, therefore, the stimulatory effect of CD on cell death is independent of its catalytic activity. Overall, our results imply that cytosolic CD stimulates apoptotic pathways by interacting with a member of the apoptotic machinery rather than by cleaving specific substrate(s).

Keywords

Lysosomal Enzyme Aspartic Protease Cell Death Differ Etoposide Treatment Aspartic Protease Inhibitor 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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References

  1. 1.
    Rochefort H (1996) The prognostic value of cathepsin D in breast cancer. A long road to the clinic. Eur J Cancer 32A:7–8.PubMedCrossRefGoogle Scholar
  2. 2.
    Liaudet-Coopman E, Beaujouin M, Derocq D, et al. (2006) Cathepsin D: newly discovered functions of a long-standing aspartic protease in cancer and apoptosis. Cancer Letters 237:167–179.PubMedCrossRefGoogle Scholar
  3. 3.
    Deiss LP, Galinka H, Berissi H, et al. (1996) Cathepsin D protease mediates programmed cell death induced by interferon gamma, Fas/APO-1 and TNF alpha. EMBO J 15:3861–3870.PubMedGoogle Scholar
  4. 4.
    Roberg K, Ollinger K (1998) Oxidative stress causes relocation of the lysosomal enzyme cathepsin D with ensuing apoptosis in neonatal rat cardiomyocytes. Am J Pathol 152:1151–1156.PubMedGoogle Scholar
  5. 5.
    Ollinger K (2000) Inhibition of cathepsin D prevents free-radical-induced apoptosis in rat cardiomyocytes. Arch Biochem Biophys 373:346–351.PubMedCrossRefGoogle Scholar
  6. 6.
    Roberg K (2001) Relocalization of cathepsin D and cytochrome c early in apoptosis revealed by immunoelectron microscopy. Lab Invest 81:149–158.PubMedGoogle Scholar
  7. 7.
    Kagedal K, Johansson U, Öllinger K (2001) The lysosomal protease cathepsin D mediates apoptosis induced by oxidative stress. FASEB J 15:1592–1594.PubMedGoogle Scholar
  8. 8.
    Takuma K, Kiriu M, Mori K, et al. (2003) Roles of cathepsins in reperfusion-induced apoptosis in cultured astrocytes. Neurochem Int 42: 153–159.PubMedCrossRefGoogle Scholar
  9. 9.
    Wu GS, Saftig P, Peters C, et al. (1998) Potential role of cathepsin D in p53-dependent tumor suppression and chemosensitivity. Oncogene 16:2177–2183.PubMedCrossRefGoogle Scholar
  10. 10.
    Emert-Sedlak L, Shangary S, Rabinovitz A, et al. (2005) Mol Cancer Ther 4:733–742.PubMedCrossRefGoogle Scholar
  11. 11.
    Johansson AC, Steen H, Öllinger K, et al. (2003) Cathepsin D mediates cytochrome c release and caspase activation in human fibroblast apoptosis induced by staurosporine. Cell Death Differ 10:1253–1259.PubMedCrossRefGoogle Scholar
  12. 12.
    Roberg K, Kagedal K, Ollinger K (2002) Microinjection of cathepsin d induces caspase-dependent apoptosis in fibroblasts. Am J Pathol 161:89–96.PubMedGoogle Scholar
  13. 13.
    Heinrich M, Neumeyer J, Jakob M, et al. (2004) Cathepsin D links TNF-induced acid sphingomyelinase to Bid-mediated caspase-9 and -3 activation. Cell Death Differ 11:550–563.PubMedCrossRefGoogle Scholar
  14. 14.
    Bidère N, Lorenzo HK, Carmona S, et al. (2003) Cathepsin D triggers Bax activation, resulting in selective apoptosis-inducing factor (AIF) relocation in T lymphocytes entering the early commitment phase to apoptosis. J Biol Chem 278: 31401–31411.PubMedCrossRefGoogle Scholar
  15. 15.
    Terman A, Neuzil J, Kagedal K, et al. (2002) Decreased apoptotic response of inclusion-cell disease fibroblasts: a consequence of lysosomal enzyme missorting? Exp Cell Res 274:9–15.PubMedCrossRefGoogle Scholar
  16. 16.
    Demoz M, Castino R, Cesaro P, et al. (2002) Endosomal-lysosomal proteolysis mediates death signalling by TNFalpha, not by etoposide, in L929 fibrosarcoma cells: evidence for an active role of cathepsin D. Biol Chem 383: 1237–1248.PubMedCrossRefGoogle Scholar
  17. 17.
    Shibata M, Kanamori S, Isahara K, et al. (1998) Participation of cathepsins B and D in apoptosis of PC12 cells following serum deprivation. Biochem Biophys Res Commun 251:199–203.PubMedCrossRefGoogle Scholar
  18. 18.
    Matsuyama S, Llopis J, Deveraux QL, et al. (2000) Changes in intramitochondrial and cytosolic pH: early events that modulate caspase activation during apoptosis. Nat Cell Biol 2:318–325.PubMedCrossRefGoogle Scholar
  19. 19.
    Capony F, Morisset M, Barrett AJ, et al. (1987) Phosphorylation, glycosylation, and proteolytic activity of the 52-kD estrogen-induced protein secreted by MCF7 cells. J Cell Biol 104:253–262.PubMedCrossRefGoogle Scholar
  20. 20.
    Tardy C, Tyynela J, Hasilik A, et al. (2003) Stress-induced apoptosis is impaired in cells with a lysosomal targeting defect but is not affected in cells synthesizing a catalytically inactive cathepsin D. Cell Death Differ 10:1090–1100.PubMedCrossRefGoogle Scholar
  21. 21.
    Reiners JJ, Jr, Caruso JA, Mathieu P, et al. (2002) Release of cytochrome c and activation of pro-caspase-9 following lysosomal photodamage involves bid cleavage. Cell Death Differ 9:934–944.PubMedCrossRefGoogle Scholar
  22. 22.
    Sawada M, Nakashima S, Banno Y, et al. (2000) Ordering of ceramide formation, caspase activation, and Bax/Bcl-2 expression during etoposide-induced apoptosis in C6 glioma cells. Cell Death Differ 7:761–772.PubMedCrossRefGoogle Scholar

Copyright information

© Springer 2008

Authors and Affiliations

  • Melanie Beaujouin
  • Emmanuelle Liaudet-Coopman
    • 1
  1. 1.Inserm Unit 540MontpellierFrance

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