Skip to main content
SpringerLink
Log in

The anti-cancer drug etoposide can induce caspase-8 processing and apoptosis in the absence of CD95 receptor-ligand interaction

  • Published:
Apoptosis Aims and scope Submit manuscript

Abstract

Caspase-8 (FLICE) can associate with and be activated by CD95 (APO-1/Fas), an apoptosis-inducing member of the Tumour Necrosis Factor receptor family. We find that, in Jurkat T cells, the DNA damaging anti-cancer drug etoposide induces apoptosis and, surprisingly, processing of caspase-8. Therefore, we have investigated whether etoposide involves CD95 receptor activation. We find that etoposide does not induce CD95 ligand expression at the mRNA level. In addition, blocking of CD95 receptor function with a specific antibody does not inhibit etoposide-induced apoptosis. Apparently, in Jurkat cells, etoposide can induce caspase-8 processing and apoptosis in a CD95-independent fashion. Likewise, we find that thymocytes from the CD95-deficient lpr/lpr mouse strain readily undergo apoptosis in response to etoposide. Moreover, since inhibition of the secretory pathway with brefeldin A does not inhibit etoposide-induced apoptosis, we exclude the requirement for a newly synthesizedreceptor ligand to induce the apoptotic pathway. We conclude that, at least in certain cell types, etoposide does not require CD95 receptor function to induce caspase-8 processing and apoptosis.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Dhein J, Walczak H, Bäumler C, Debatin K-M, Krammer PH. Autocrine T-cell suicide mediated by APO-1/(fas/CD95). Nature 1995; 373: 438-441.

    Google Scholar 

  2. Brunner T, Mogli RJ, LaFace D, et al. Cell-autonomous Fas (CD95)/Fas-ligand interaction mediates activation-induced apoptosis in T cell hybridomas. Nature 1995; vn 373: 441-444.

    Google Scholar 

  3. Ju S-T, Panka DJ, Cul H, et al. Fas (CD95)/FasL interactions required for programmed cell death after T-cell activation. Nature 1995; 373: 444-448.

    Google Scholar 

  4. Russell JH, Rush B, Weaver C, Wang R. Mature T cells of the autoimmune lpr/lpr mice have a defect in antigen-stimulated suicide. Proc Natl Acad Sci USA 1993; 90: 4409-4413.

    Google Scholar 

  5. Singer GC, Abbas A. The Fas antigen is involved in peripheral but not thymic deletion of T lymphocytes in T cell receptor transgenic mice. Immunity 1994; 1: 365-371.

    Google Scholar 

  6. Friesen C, Herr I, Krammer PH, Debatin K-M. Involvement of the CD95 (APO-1/Fas) receptor/ligand system in drug-induced apoptosis in leukemia cells. Nature Med 1996; 2: 574-577.

    Google Scholar 

  7. Müller M, Strand S, Hug H, et al. Drug-induced apoptosis in hepatoma cells is mediated by the CD95 (APO-1/Fas) receptor/ligand system and involves activation of wild type p53. J Clin Invest 1997; 99: 403-413.

    Google Scholar 

  8. Chinnaiyan AM, Dixit VM. The cell-death machine. Curr Biol 1996; 6: 555-562.

    Google Scholar 

  9. Nicholson D, Thornberry NA. Caspases: killer proteases. TIBS 1997; 22: 299-306.

    Google Scholar 

  10. Chinnaiyan AM, O'Rourke K, Tewari M, Dixit VM. FADD, a novel death domain-containing protein, interacts with the death domain of Fas and initiates apoptosis. Cell 1995; 81: 505-512.

    Google Scholar 

  11. Muzio M, Chinnaiyan AM, Kischkel FC, et al. FLICE, a novel FADD-homologous ICE/CED-3-like protease, is recruited to the CD95 (Fas/APO-1) death-inducing signaling complex. Cell 1996; 85: 817-827.

    Google Scholar 

  12. Boldin MP, Goncharov TM, Goltsev YV, Wallach D. Involvement of MACH, a novel MORT1/FADD-interacting protease, in Fas/APO-1-and TNF receptor-induced cell death. Cell 1996; 85: 803-815.

    Google Scholar 

  13. Fernandes-Alnemri T, Armstrong RC, Krebs J, et al. In vitro activation of CPP32 and Mch3 by Mch4, a novel human apoptotic cysteine protease containing two FADD-like domains. Proc Natl Acad Sci USA 1996; 93: 7464-7469.

    Google Scholar 

  14. Chinnaiyan AM, Orth K, O'Rourke K, Duan H, Poirier GG, Dixit VM. Molecular ordering of the cell death pathway. J Biol Chem 1996; 271: 4573-4576.

    Google Scholar 

  15. Schlegel J, Peters I, Orrenius S, et al. CPP-32/Apopain is the key interleukin-1β-converting enzyme-like protease involved in Fas-mediated apoptosis. J Biol Chem 1996; 271: 1841-1844.

    Google Scholar 

  16. Srinivasula SM, Ahmad M, Fernandes-Alnemri T, Litwack G, Alnemri ES. Molecular ordering of the Fas-apoptotic pathway: The Fas/APO-1 protease Mch5 is a CrmA-inhibitable protease that activates multiple Ced-3/ICE-like cysteine proteases. Proc Natl Acad Sci USA 1996; 93: 14486-14491.

    Google Scholar 

  17. Nelson EM, Tewey KM, Liu LF. Mechanism of antitumor drug action: poisoning of mammalian DNA topoisomerase II on DNA by 4'-(9-acridinylamino)-methanesulfon-m-anisidide. Proc Natl Acad Sci USA 1984; 81: 1361-1365.

    Google Scholar 

  18. Chilson OP, Boylston AW, Crumpton MJ. Phaseolus vulgaris phytohaemagglutinin (PHA) binds to the human T lymphocyte antigen receptor. EMBO J 1984; vn 3: 3239-3245.

    Google Scholar 

  19. Ogasawara J, Watenabe-Fukunaga R, Adachi M, et al. Lethal effects of the anti-Fas antibody in mice. Nature 1993; 364: 806-809.

    Google Scholar 

  20. Watanabe-Fukunaga R, Brannan C, Copeland NG, Jenkins NA, Nagata S. Lympho-proliferation disorder in mice explained by defects in Fas antigen that mediates apoptosis. Nature 1992; 356: 314-317.

    Google Scholar 

  21. Yssel H, de Vries JE, Koken M, van Blitterswijk WJ, Spits H. J Immunol Meth 1984; 72: 219-227.

    Google Scholar 

  22. Nicoletti I, Migliorati G, Pagliacci MC, Grignani F, Riccardi C. A rapid and simple method for measuring thymocyte apoptosis by propidium iodide staining and flow cytometry. J Immunol Meth 1991; 139: 271-279.

    Google Scholar 

  23. Vrieling H, Niericker MJ, Simons JWIM, van Zeeland AA. Molecular analysis of mutations induced by N-ethyl-N-nitrosourea at HPRT locus in mouse lymphoma cells. Mut Res 1988; 198: 99-106.

    Google Scholar 

  24. Schuuring E, Verhoeven E, Mooi WJ, Michalides RJAM. Identification and cloning of two overexpressed genes U21B31/PRAD1 and EMS1, within the amplified chromosome 11q13 region in human carcinomas. Oncogene 1992; 7: 355-361.

    Google Scholar 

  25. Medema J-P, Scaffidi C, Kischkel F, et al. FLICE is activated by association with the CD95 death-inducing signaling complex (DISC). EMBO J 1997; 16: 2794-2804.

    Google Scholar 

  26. Dubrez L, Savoy I, Hamman A, Solary E. Pivotal role of a DEVD-sensitive step in etoposide-induced and Fasmediated apoptotic pathways. EMBO J 1996; 15: 5504-5512.

    Google Scholar 

  27. Erhardt P, Cooper G. Activation of the CPP32 apoptotic protease by distinct signaling pathways with differential sensitivity to Bcl-xL. J Biol Chem 1996; 271: 17601-17604.

    Google Scholar 

  28. Martins LM, Kottke T, Mesner PW, et al. Activation of multiple interleukin-1β converting enzyme homologues in cytosol and nuclei of HL-60 cells during etoposide-induced apoptosis. J Biol Chem 1997; 272: 7421-7430.

    Google Scholar 

  29. Pan G, O'Rourke K, Chinnaiyan AM, et al. The receptor for the cytotoxic ligand TRAIL. Science 1997; 276: 111-113.

    Google Scholar 

  30. Chinnayian AM, O'Rourke K, Yu G-L, et al. Signal transduction by DR3, a death domain-containing receptor related to TNFR-1 and CD95. Science 1996; 274: 990-992.

    Google Scholar 

  31. Kitson J, Raven T, Jiang Y-P, et al. A death-domaincontaining receptor that mediates apoptosis. Nature 1996; 384: 372-375.

    Google Scholar 

  32. Marsters SA, Sheridan JP, Donahue CJ, et al. Apo-3, a new member of the tumor necrosis factor receptor family, contains a death domain and activates apoptosis and NF-kB. Curr Biol 1996; 6: 1669-1676.

    Google Scholar 

  33. Bodmer J-L, Burns K, Schneider P, et al. TRAMP, a novel apoptosis-mediating receptor with sequence homology to Tumor Necrosis Factor Receptor 1 and Fas(Apo-1/CD95). Immunity 1997; 6: 79-88.

    Google Scholar 

  34. Fujiwara T, Oda K, Takasuki A, Ikehara Y. Brefeldin A causes disassembly of the Golgi complex and accumulation of secretory proteins in the endoplasmic reticulum. J Biol Chem 1988; 263: 18545-18552.

    Google Scholar 

  35. Tewari M, Quan LT, O'Rourke K, et al. Yama/CPP32b a mammalian homologue of CED-3 is a CrmA-inhibitable protease that cleaves the death substrate poly(ADP-ribose) polymerase. Cell 1995; 81: 801-809.

    Google Scholar 

  36. Clarke AR, Purdie CA, Harrison DJ, Morris RG, Bird CC, Hooper ML, Wyllie AH. Thymocyte apoptosis induced by p53-dependent and independent pathways. Nature 1993; 362: 849-852.

    Google Scholar 

  37. Miyashita T, Reed JC. 1995. Tumor suppressor p53 is a direct transcriptional activator of the human bax gene. Cell 1995; 80: 293-299.

    Google Scholar 

  38. Brady HJM, Salamons GS, Bobeldijk RC, Berns AJM. T cells from baxa transgenic mice show accelerated apoptosis in response to stimuli but do not show restored DNA damage-induced death in the absence of p53. EMBO J 1996; 15: 1221-1230.

    Google Scholar 

  39. Owen-Schaub LB, Zhang W, Cusak JC, et al. Wild-type human p53 and a temperature-sensitive mutant induce Fas/APO-1 expression. Mol Cell Biol 1995; 15: 3032-3040.

    Google Scholar 

  40. Strasser A, Harris AW, Huang DCS, Krammer PH, Cory S. Bcl-2 and Fas/APO-1 regulate distinct pathways to lymphocyte apoptosis. EMBO J 1995; 14: 6136-6147.

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Division of Cellular Biochemistry, The Netherlands Cancer Institute, 1066 CX, Amsterdam, The Netherlands

    J. G. R. Boesen-de Cock, E. de Vries & J. Borst

  2. Department of Biological Sciences, Keele University, Keele, UK

    G. T. Williams

Authors
  1. J. G. R. Boesen-de Cock
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. E. de Vries
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. G. T. Williams
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. J. Borst
    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

Boesen-de Cock, J.G.R., de Vries, E., Williams, G.T. et al. The anti-cancer drug etoposide can induce caspase-8 processing and apoptosis in the absence of CD95 receptor-ligand interaction. Apoptosis 3, 17–25 (1998). https://doi.org/10.1023/A:1009603001888

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1009603001888

Search

Navigation