Skip to main content
SpringerLink
Log in

Doxorubicin enhances TRAIL-induced cell death via ceramide-enriched membrane platforms

  • Original Paper
  • Published:
Apoptosis Aims and scope Submit manuscript

Abstract

Previous studies indicated that signalling via CD95 and DR5 is greatly enhanced by the formation of ceramide-enriched membrane platforms. Here, we employed this concept to convert doses of subtherapeutic TRAIL that were unable to release ceramide and kill leukemic B-cells or ex vivo T lymphocytes, into a very effective apoptotic stimulus. Ceramide production was induced by application of sub-toxic doses of doxorubicin that resulted in an activation of the acid sphingomyelinase (ASM), release of ceramide and formation of ceramide-enriched membrane platforms. The latter served DR5 to cluster after application of very low doses of TRAIL in combination with doxorubicin. Genetic deficiency of the ASM abrogated doxorubicin-induced ceramide release, as well as clustering of DR5 and apoptosis induced by the combined treatment of doxorubicin and TRAIL. These data show that local release of ceramide potentiates very low, otherwise inactive doses of TRAIL that may represent a novel therapeutic concept to treat tumors.

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

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  1. Wiley SR, Schooley K, Smolak PJ, Din WS, Huang CP, Nicholl JK, Sutherland GR, Smith TD, Rauch C, Smith CA, et al (1995) Identification and characterization of a new member of the TNF family that induces apoptosis. Immunity 3:673–682

    Article  PubMed  CAS  Google Scholar 

  2. Pan G, O’Rourke K, Chinnaiyan AM, Gentz R, Ebner R, Ni J, Dixit VM (1997) The receptor for the cytotoxic ligand TRAIL. Science 276:111–113

    Article  PubMed  CAS  Google Scholar 

  3. Sheridan JP, Marsters SA, Pitti RM, Gurney A, Skubatch M, Baldwin D, Ramakrishnan L, Gray CL, Baker K, Wood WI, Goddard AD, Godowski P, Ashkenazi A (1997) Control of TRAIL-induced apoptosis by a family of signaling and decoy receptors. Science 277:818–821

    Article  PubMed  CAS  Google Scholar 

  4. Hayakawa Y, Screpanti V, Yagita H, Grandien A, Ljunggren HG, Smyth MJ, Chambers BJ (2004) NK cell TRAIL eliminates immature dendritic cells in vivo and limits dendritic cell vaccination efficacy. J Immunol 172:123–129

    PubMed  CAS  Google Scholar 

  5. Janssen EM, Droin NM, Lemmens EE, Pinkoski MJ, Bensinger SJ, Ehst BD, Griffith TS, Green DR, Schoenberger SP (2005) CD4+ T-cell help controls CD8+ T-cell memory via TRAIL-mediated activation-induced cell death. Nature 434:88–93

    Article  PubMed  CAS  Google Scholar 

  6. Ursini-Siegel J, Zhang W, Altmeyer A, Hatada EN, Do RK, Yagita H, Chen-Kiang S (2002) TRAIL/Apo-2 ligand induces primary plasma cell apoptosis. J Immunol 169:5505–5513

    PubMed  CAS  Google Scholar 

  7. Zhang L, Gu J, Lin T, Huang X, Roth JA, Fang B (2002) Mechanisms involved in development of resistance to adenovirus-mediated proapoptotic gene therapy in DLD1 human colon cancer cell line. Gene Ther 9:1262–1270

    Article  PubMed  CAS  Google Scholar 

  8. Duiker EW, Mom CH, de Jong S, Willemse PH, Gietema JA, van der Zee AG, de Vries EG (2006) The clinical trail of TRAIL. Eur J Cancer 42:2233–2240

    Article  PubMed  CAS  Google Scholar 

  9. Pan G, Ni J, Wei YF, Yu G, Gentz R, Dixit VM (1997) An antagonist decoy receptor and a death domain-containing receptor for TRAIL. Science 277:815–818

    Article  PubMed  CAS  Google Scholar 

  10. Schneider P, Thome M, Burns K, Bodmer JL, Hofmann K, Kataoka T, Holler N, Tschopp J (1997) TRAIL receptors 1 (DR4) and 2 (DR5) signal FADD-dependent apoptosis and activate NF-kappaB. Immunity 7:831–836

    Article  PubMed  CAS  Google Scholar 

  11. Wu GS, Burns TF, Zhan Y, Alnemri ES, El-Deiry WS (1999) Molecular cloning and functional analysis of the mouse homologue of the KILLER/DR5 tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) death receptor. Cancer Res 59:2770–2775

    PubMed  CAS  Google Scholar 

  12. Degli-Esposti MA, Smolak PJ, Walczak H, Waugh J, Huang CP, DuBose RF, Goodwin RG, Smith CA (1997) Cloning and characterization of TRAIL-R3, a novel member of the emerging TRAIL receptor family. J Exp Med 186:1165–1170

    Article  PubMed  CAS  Google Scholar 

  13. Emery JG, McDonnell P, Burke MB, Deen KC, Lyn S, Silverman C, Dul E, Appelbaum ER, Eichman C, DiPrinzio R, Dodds RA, James IE, Rosenberg M, Lee JC, Young PR (1998) Osteoprotegerin is a receptor for the cytotoxic ligand TRAIL. J Biol Chem 273:14363–14367

    Article  PubMed  CAS  Google Scholar 

  14. Simons K, Ikonen E (1997) Functional rafts in cell membranes. Nature 387:569–572

    Article  PubMed  CAS  Google Scholar 

  15. Harder T, Simons K (1997) Caveolae, DIGs, and the dynamics of sphingolipid-cholesterol microdomains. Curr Opin Cell Biol 9:534–542

    Article  PubMed  CAS  Google Scholar 

  16. Brown DA, London E (1998) Functions of lipid rafts in biological membranes. Ann Rev Cell Dev Biol 14:111–136

    Article  CAS  Google Scholar 

  17. Dumitru CA, Gulbins E (2006) TRAIL activates acid sphingomyelinase via a redox mechanism and releases ceramide to trigger apoptosis. Oncogene 25:5612–5625

    Article  PubMed  CAS  Google Scholar 

  18. Walczak H, Degli-Esposti MA, Johnson RS, Smolak PJ, Waugh JY, Boiani N, Timour MS, Gerhart MJ, Schooley KA, Smith CA, Goodwin RG, Rauch CT (1997) TRAIL-R2: a novel apoptosis-mediating receptor for TRAIL. Embo J 16:5386–5397

    Article  PubMed  CAS  Google Scholar 

  19. Mariani SM, Matiba B, Armandola EA, Krammer PH (1997) Interleukin 1 beta-converting enzyme related proteases/caspases are involved in TRAIL-induced apoptosis of myeloma and leukemia cells. J Cell Biol 137:221–229

    Article  PubMed  CAS  Google Scholar 

  20. Obeid LM, Linardic CM, Karolak LA, Hannun YA (1993) Programmed cell death induced by ceramide. Science 259:1769–1771

    Article  PubMed  CAS  Google Scholar 

  21. Cifone MG, De Maria R, Roncaioli P, Rippo MR, Azuma M, Lanier LL, Santoni A, Testi R (1994) Apoptotic signaling through CD95 (Fas/Apo-1) activates an acidic sphingomyelinase. J Exp Med 180:1547–1552

    Article  PubMed  CAS  Google Scholar 

  22. Gulbins E, Bissonnette R, Mahboubi A, Martin S, Nishioka W, Brunner T, Baier G, Baier-Bitterlich G, Byrd C, Lang F, et al (1995) FAS-induced apoptosis is mediated via a ceramide-initiated RAS signaling pathway. Immunity 2:341–351

    Article  PubMed  CAS  Google Scholar 

  23. Grassme H, Jekle A, Riehle A, Schwarz H, Berger J, Sandhoff K, Kolesnick R, Gulbins E (2001) CD95 signaling via ceramide-rich membrane rafts. J Biol Chem 276:20589–20596

    Article  PubMed  CAS  Google Scholar 

  24. Grassme H, Schwarz H, Gulbins E (2001) Molecular mechanisms of ceramide-mediated CD95 clustering. Biochem Biophys Res Commun 284:1016–1030

    Article  PubMed  CAS  Google Scholar 

  25. Schutze S, Potthoff K, Machleidt T, Berkovic D, Wiegmann K, Kronke M (1992) TNF activates NF-kappa B by phosphatidylcholine-specific phospholipase C-induced “acidic” sphingomyelin breakdown. Cell 71:765–776

    Article  PubMed  CAS  Google Scholar 

  26. Zhang Y, Mattjus P, Schmid PC, Dong Z, Zhong S, Ma WY, Brown RE, Bode AM, Schmid HH, Dong Z (2001) Involvement of the acid sphingomyelinase pathway in uva-induced apoptosis. J Biol Chem 276:11775–11782

    Article  PubMed  CAS  Google Scholar 

  27. Haimovitz-Friedman A, Kan CC, Ehleiter D, Persaud RS, McLoughlin M, Fuks Z, Kolesnick RN (1994) Ionizing radiation acts on cellular membranes to generate ceramide and initiate apoptosis. J Exp Med 180:525–535

    Article  PubMed  CAS  Google Scholar 

  28. Lacour S, Hammann A, Grazide S, Lagadic-Gossmann D, Athias A, Sergent O, Laurent G, Gambert P, Solary E, Dimanche-Boitrel MT (2004) Cisplatin-induced CD95 redistribution into membrane lipid rafts of HT29 human colon cancer cells. Cancer Res 64:3593–3598

    Article  PubMed  CAS  Google Scholar 

  29. Modrak DE, Cardillo TM, Newsome GA, Goldenberg DM, Gold DV (2004) Synergistic interaction between sphingomyelin and gemcitabine potentiates ceramide-mediated apoptosis in pancreatic cancer. Cancer Res 64:8405–8410

    Article  PubMed  CAS  Google Scholar 

  30. Lovat PE, Di Sano F, Corazzari M, Fazi B, Donnorso RP, Pearson AD, Hall AG, Redfern CP, Piacentini M (2004) Gangliosides link the acidic sphingomyelinase-mediated induction of ceramide to 12-lipoxygenase-dependent apoptosis of neuroblastoma in response to fenretinide. J Natl Cancer Inst 96:1288–1299

    Article  PubMed  CAS  Google Scholar 

  31. Fulda S, Meyer E, Friesen C, Susin SA, Kroemer G, Debatin KM (2001) Cell type specific involvement of death receptor and mitochondrial pathways in drug-induced apoptosis. Oncogene 20:1063–1075

    Article  PubMed  CAS  Google Scholar 

  32. Jendrossek V, Muller I, Eibl H, Belka C (2003) Intracellular mediators of erucylphosphocholine-induced apoptosis. Oncogene 22:2621–2631

    Article  PubMed  CAS  Google Scholar 

  33. Gulbins E, Kolesnick R (2000) Measurement of sphingomyelinase activity. Methods Enzymol 322:382–388

    PubMed  CAS  Google Scholar 

  34. Perry DK, Bielawska A, Hannun YA (2000) Quantitative determination of ceramide using diglyceride kinase. Methods Enzymol 312:22–31

    Article  PubMed  CAS  Google Scholar 

  35. Morita Y, Perez GI, Paris F, Miranda SR, Ehleiter D, Haimovitz-Friedman A, Fuks Z, Xie Z, Reed JC, Schuchman EH, Kolesnick RN, Tilly JL (2000) Oocyte apoptosis is suppressed by disruption of the acid sphingomyelinase gene or by sphingosine-1-phosphate therapy. Nat Med 6:1109–1114

    Article  PubMed  CAS  Google Scholar 

  36. Andrieu-Abadie N, Jaffrezou JP, Hatem S, Laurent G, Levade T, Mercadier JJ (1999) L-carnitine prevents doxorubicin-induced apoptosis of cardiac myocytes: role of inhibition of ceramide generation. Faseb J 13:1501–1510

    PubMed  CAS  Google Scholar 

  37. Gouaze V, Mirault ME, Carpentier S, Salvayre R, Levade T, Andrieu-Abadie N (2001) Glutathione peroxidase-1 overexpression prevents ceramide production and partially inhibits apoptosis in doxorubicin-treated human breast carcinoma cells. Mol Pharmacol 60:488–496

    PubMed  CAS  Google Scholar 

  38. Mercier C, Decleves X, Masseguin C, Fragner P, Tardy M, Roux F, Gabrion J, Scherrmann JM (2003) P-glycoprotein (ABCB1) but not multidrug resistance-associated protein 1 (ABCC1) is induced by doxorubicin in primary cultures of rat astrocytes. J Neurochem 87:820–830

    Article  PubMed  CAS  Google Scholar 

  39. Cremesti A, Paris F, Grassme H, Holler N, Tschopp J, Fuks Z, Gulbins E, Kolesnick R (2001) Ceramide enables fas to cap and kill. J Biol Chem 276:23954–23961

    Article  PubMed  CAS  Google Scholar 

  40. Fanzo JC, Lynch MP, Phee H, Hyer M, Cremesti A, Grassme H, Norris JS, Coggeshall KM, Rueda BR, Pernis AB, Kolesnick R, Gulbins E (2003) CD95 rapidly clusters in cells of diverse origins. Cancer Biol Ther 2:392–395

    PubMed  CAS  Google Scholar 

  41. Nachbur U, Kassahn D, Yousefi S, Legler DF, Brunner T (2006) Posttranscriptional regulation of Fas (CD95) ligand killing activity by lipid rafts. Blood 107:2790–2796

    Article  PubMed  CAS  Google Scholar 

  42. Boldin MP, Mett IL, Varfolomeev EE, Chumakov I, Shemer-Avni Y, Camonis JH, Wallach D (1995) Self-association of the “death domains” of the p55 tumor necrosis factor (TNF) receptor and Fas/APO1 prompts signaling for TNF and Fas/APO1 effects. J Biol Chem 270:387–391

    Article  PubMed  CAS  Google Scholar 

  43. Grassme H, Bock J, Kun J, Gulbins E (2002) Clustering of CD40 ligand is required to form a functional contact with CD40. J Biol Chem 277:30289–30299

    Article  PubMed  CAS  Google Scholar 

  44. Grassme H, Jendrossek V, Bock J, Riehle A, Gulbins E (2002) Ceramide-rich membrane rafts mediate CD40 clustering. J Immunol 168:298–307

    PubMed  CAS  Google Scholar 

  45. Martin S, Phillips DC, Szekely-Szucs K, Elghazi L, Desmots F, Houghton JA (2005) Cyclooxygenase-2 inhibition sensitizes human colon carcinoma cells to TRAIL-induced apoptosis through clustering of DR5 and concentrating death-inducing signaling complex components into ceramide-enriched caveolae. Cancer Res 65:11447–11458

    Article  PubMed  CAS  Google Scholar 

  46. Bock J, Gulbins E (2003) The transmembranous domain of CD40 determines CD40 partitioning into lipid rafts. FEBS Lett 534:169–174

    Article  PubMed  CAS  Google Scholar 

  47. Qiu H, Edmunds T, Baker-Malcolm J, Karey KP, Estes S, Schwarz C, Hughes H, Van Patten SM (2003) Activation of human acid sphingomyelinase through modification or deletion of C-terminal cysteine. J Biol Chem 278:32744–32752

    Article  PubMed  CAS  Google Scholar 

  48. Reinehr R, Becker S, Braun J, Eberle A, Grether-Beck S, Haussinger D (2006) Endosomal acidification and activation of NADPH oxidase isoforms are upstream events in hyperosmolarity-induced hepatocyte apoptosis. J Biol Chem 281:23150–23166

    Article  PubMed  CAS  Google Scholar 

  49. Sinha BK, Mimnaugh EG, Rajagopalan S, Myers CE (1989) Adriamycin activation and oxygen free radical formation in human breast tumor cells: protective role of glutathione peroxidase in adriamycin resistance. Cancer Res 49:3844–3848

    PubMed  CAS  Google Scholar 

  50. Ubezio P, Civoli F (1994) Flow cytometric detection of hydrogen peroxide production induced by doxorubicin in cancer cells. Free Radic Biol Med 16:509–516

    Article  PubMed  CAS  Google Scholar 

  51. Cervantes A, Pinedo HM, Lankelma J, Schuurhuis GJ (1988) The role of oxygen-derived free radicals in the cytotoxicity of doxorubicin in multidrug resistant and sensitive human ovarian cancer cells. Cancer Lett 41:169–177

    Article  PubMed  CAS  Google Scholar 

  52. Ciusani E, Croci D, Gelati M, Calatozzolo C, Sciacca F, Fumagalli L, Balzarotti M, Fariselli L, Boiardi A, Salmaggi A (2005) In vitro effects of topotecan and ionizing radiation on TRAIL/Apo2L-mediated apoptosis in malignant glioma. J Neurooncol 71:19–25

    Article  PubMed  CAS  Google Scholar 

  53. Shankar S, Chen X, Srivastava RK (2005) Effects of sequential treatments with chemotherapeutic drugs followed by TRAIL on prostate cancer in vitro and in vivo. Prostate 62:165–186

    Article  PubMed  CAS  Google Scholar 

  54. Gibson SB, Oyer R, Spalding AC, Anderson SM, Johnson GL (2000) Increased expression of death receptors 4 and 5 synergizes the apoptosis response to combined treatment with etoposide and TRAIL. Mol Cell Biol 20:205–212

    Article  PubMed  CAS  Google Scholar 

  55. Ballestrero A, Nencioni A, Boy D, Rocco I, Garuti A, Mela GS, Van Parijs L, Brossart P, Wesselborg S, Patrone F (2004) Tumor necrosis factor-related apoptosis-inducing ligand cooperates with anticancer drugs to overcome chemoresistance in antiapoptotic Bcl-2 family members expressing jurkat cells. Clin Cancer Res 10:1463–1470

    Article  PubMed  CAS  Google Scholar 

  56. Jin H, Yang R, Fong S, Totpal K, Lawrence D, Zheng Z, Ross J, Koeppen H, Schwall R, Ashkenazi A (2004) Apo2 ligand/tumor necrosis factor-related apoptosis-inducing ligand cooperates with chemotherapy to inhibit orthotopic lung tumor growth and improve survival. Cancer Res 64:4900–4905

    Article  PubMed  CAS  Google Scholar 

  57. Marini P, Denzinger S, Schiller D, Kauder S, Welz S, Humphreys R, Daniel PT, Jendrossek V, Budach W, Belka C (2006) Combined treatment of colorectal tumours with agonistic TRAIL receptor antibodies HGS-ETR1 and HGS-ETR2 and radiotherapy: enhanced effects in vitro and dose-dependent growth delay in vivo. Oncogene 25:5145–5154

    PubMed  CAS  Google Scholar 

  58. Marini P, Schmid A, Jendrossek V, Faltin H, Daniel PT, Budach W, Belka C (2005) Irradiation specifically sensitises solid tumour cell lines to TRAIL mediated apoptosis. BMC Cancer 5:5

    Article  PubMed  CAS  Google Scholar 

  59. Thai le M, Labrinidis A, Hay S, Liapis V, Bouralexis S, Welldon K, Coventry BJ, Findlay DM, Evdokiou A (2006) Apo2l/Tumor necrosis factor-related apoptosis-inducing ligand prevents breast cancer-induced bone destruction in a mouse model. Cancer Res 66:5363–5370

    Article  PubMed  CAS  Google Scholar 

  60. Straughn JM Jr, Oliver PG, Zhou T, Wang W, Alvarez RD, Grizzle WE, Buchsbaum DJ (2006) Anti-tumor activity of TRA-8 anti-death receptor 5 (DR5) monoclonal antibody in combination with chemotherapy and radiation therapy in a cervical cancer model. Gynecol Oncol 101:46–54

    Article  PubMed  CAS  Google Scholar 

  61. Cheong I, Huang X, Bettegowda C, Diaz LA Jr, Kinzler KW, Zhou S, Vogelstein B (2006) A bacterial protein enhances the release and efficacy of liposomal cancer drugs. Science 314:1308–1311

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgements

The study was supported by DFG Graduiertenkolleg 1045/1 to E.G. and Deutsche Krebshilfe grant 102238-Gu2 to T.T., U.H. and E.G.

Author information

Authors and Affiliations

  1. Department of Molecular Biology, University of Duisburg-Essen, Hufelandstrasse 55, 45122, Essen, Germany

    Claudia Alexandra Dumitru, Alexander Carpinteiro & Erich Gulbins

  2. Clinic of Hematology, University of Duisburg-Essen, 45122, Essen, Germany

    Alexander Carpinteiro

  3. Department of Internal Medicine/Hematology Oncology (West German Cancer Center), University of Duisburg-Essen, 45122, Essen, Germany

    Tanja Trarbach

  4. Department of Dermatology, University of Duesseldorf, 40225, Duesseldorf, Germany

    Ulrich R. Hengge

Authors
  1. Claudia Alexandra Dumitru
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Alexander Carpinteiro
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Tanja Trarbach
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Ulrich R. Hengge
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Erich Gulbins
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Erich Gulbins.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Dumitru, C.A., Carpinteiro, A., Trarbach, T. et al. Doxorubicin enhances TRAIL-induced cell death via ceramide-enriched membrane platforms. Apoptosis 12, 1533–1541 (2007). https://doi.org/10.1007/s10495-007-0081-9

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10495-007-0081-9

Keywords

Search

Navigation