Skip to main content
SpringerLink
Log in

CD95/Apo-1/Fas: independent cell death induced by doxorubicin in normal cultured cardiomyocytes

  • Original Article
  • Published:
Cancer Immunology, Immunotherapy Aims and scope Submit manuscript

Abstract

Doxorubicin is a commonly used cytotoxic drug for effective treatment of both solid tumors and leukemias, which may cause severe cardiac adverse effects leading to heart failure. In certain tumor cells, doxorubicin-induced cell death is mediated by death receptors such as CD95/Apo-1/Fas. Here we studied the role of death receptors for doxorubicin-induced cell death in primary neonatal rat cardiomyocytes and the embryonic cardiomyocytic cell line H9c2.1. Doxorubicin-induced cell death of cardiomyocytes was associated with cleavage of caspases 3 and 8, a drop in mitochondrial transmembrane potential, and release of cytochrome c. Doxorubicin-treated cardiomyocytes secreted death-inducing ligands into the culture supernatant, but remained resistant toward cell death induction by death receptor triggering. In contrast to the chelator dexrazoxane, blockade of death receptor signaling by stable overexpression of transdominant negative adapter molecule FADD did not inhibit doxorubicin-induced cell death. Our data suggest that cultured cardiomyocytes secrete death-inducing ligands, but undergo death receptor–independent cell death upon exposure to doxorubicin.

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. Arola OJ, Saraste A, Pulkki K, Kallajoki M, Parvinen M, Voipio-Pulkki LM (2000) Acute doxorubicin cardiotoxicity involves cardiomyocyte apoptosis. Cancer Res 60:1789

    CAS  PubMed  Google Scholar 

  2. Bergmann MW, Loser P, Dietz R, von Harsdorf R (2001) Effect of NF-kappa B inhibition on TNF-alpha-induced apoptosis and downstream pathways in cardiomyocytes. J Mol Cell Cardiol 33:1223

    Article  CAS  PubMed  Google Scholar 

  3. Bonavita F, Stefanelli C, Giordano E, Columbaro M, Facchini A, Bonafe F, Caldera CM, Guaneri C (2003) H9c2 cardiac myoblasts undergo apoptosis in a model of ischemia consisting of serum deprivation and hypoxia: inhibition by PMA. FEBS Lett 536:85

    Article  CAS  PubMed  Google Scholar 

  4. Chao W, Shen Y, Li L, Rosenzweig A (2002) Importance of FADD signaling in serum-deprivation- and hypoxia-induced cardiomyocyte apoptosis. J Biol Chem 277:31639

    Google Scholar 

  5. Childs AC, Phaneuf SL, Dirks AJ, Phillips T, Leeuwenburgh C (2002) Doxorubicin treatment in vivo causes cytochrome C release and cardiomyocyte apoptosis, as well as increased mitochondrial efficiency, superoxide dismutase activity, and Bcl-2:Bax ratio. Cancer Res 62:4592

    CAS  PubMed  Google Scholar 

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

    Article  CAS  PubMed  Google Scholar 

  7. Clementi ME, Giardina B, Di Stasio E, Mordente A, Misiti F (2003) Doxorubicin-derived metabolites induce release of cytochrome C and inhibition of respiration on cardiac isolated mitochondria. Anticancer Res 23:2445

    CAS  PubMed  Google Scholar 

  8. Cusack BJ, Young SP, Driskell J, Olson RD (1993) Doxorubicin and doxorubicinol pharmacokinetics and tissue concentrations following bolus injection and continuous infusion of doxorubicin in the rabbit. Cancer Chemother Pharmacol 32:53

    CAS  PubMed  Google Scholar 

  9. Friesen C, Herr I, Krammer PH, Debatin KM (1996) Involvement of the CD95 (APO-1/Fas) receptor/ligand system in drug-induced apoptosis in leukemia cells. Nat Med 2:574

    Article  CAS  PubMed  Google Scholar 

  10. Friesen C, Fulda S, Debatin KM (1999) Induction of CD95 ligand and apoptosis by doxorubicin is modulated by the redox state in chemosensitive- and drug-resistant tumor cells. Cell Death Differ 6:471

    Article  CAS  PubMed  Google Scholar 

  11. Fulda S, Sieverts H, Friesen C, Herr I, Debatin KM (1997) The CD95 (APO-1/Fas) system mediates drug-induced apoptosis in neuroblastoma cells. Cancer Res 57:3823

    CAS  PubMed  Google Scholar 

  12. Hasinoff BB (1989) The interaction of the cardioprotective agent ICRF-187 [+]-1,2-bis(3,5-dioxopiperazinyl-1-yL)propane; its hydrolysis product (ICRF-198); and other chelating agents with the Fe(III) and Cu(II) complexes of adriamycin. Agents Actions 26:378

    CAS  PubMed  Google Scholar 

  13. Herr I, Debatin KM (2001) Cellular stress response and apoptosis in cancer therapy. Blood 98:2603

    Article  CAS  PubMed  Google Scholar 

  14. Janin A, Deschaumes C, Daneshpouy M, Estaquier J, Micic-Polianski J, Rajagopalan-Levasseur P, Akarid K, Mounier N, Gluckman E, Socie G, Ameisen JC (2002) CD95 engagement induces disseminated endothelial cell apoptosis in vivo: immunopathologic implications. Blood 99:2940

    Article  CAS  PubMed  Google Scholar 

  15. Jemmerson R, Liu J, Hausauer D, Lam KP, Mondino A, Nelson RD (1999) A conformational change in cytochrome c of apoptotic and necrotic cells is detected by monoclonal antibody binding and mimicked by association of the native antigen with synthetic phospholipid vesicles. Biochemistry 38:3599

    Article  CAS  PubMed  Google Scholar 

  16. Jeremias I, Herr I, Boehler T, Debatin KM (1998) TRAIL/Apo-2-Ligand induced apoptosis in T-cells. Eur J Immunol 28:143

    Google Scholar 

  17. Jeremias I, Kupatt C, Martin-Villalba A, Habazettl H, Schenkel J, Boekstegers P, Debatin KM (2000) Involvement of CD95/Apo1/Fas in cell death after myocardial ischemia. Circulation 102:915

    Google Scholar 

  18. Kayagaki N, Kawasaki A, Ebata T, Ohmoto H, Ikeda S, Inoue S, Yoshino K, Okumura K, Yagita H (1995) Metalloproteinase-mediated release of human Fas ligand. J Exp Med 182:1777

    Article  CAS  PubMed  Google Scholar 

  19. Keefe DL (2001) Anthracycline-induced cardiomyopathy. Semin Oncol 28:2

    Article  CAS  Google Scholar 

  20. Kumar D, Kirshenbaum LA, Li T, Danelisen I, Singal PK (2001) Apoptosis in adriamycin cardiomyopathy and its modulation by probucol. Antioxid Redox Signal 3:135

    Article  CAS  PubMed  Google Scholar 

  21. Lebrecht D, Setzer B, Ketelsen UP, Haberstroh J, Walker UA (2003) Time-dependent and tissue-specific accumulation of mtDNA and respiratory chain defects in chronic doxorubicin cardiomyopathy. Circulation 108:2423

    Google Scholar 

  22. Nakamura T, Ueda Y, Juan Y, Katsuda S, Takahashi H, Koh E (2000) Fas-mediated apoptosis in adriamycin-induced cardiomyopathy in rats: in vivo study. Circulation 102:572

    Google Scholar 

  23. Newton K, Strasser A (2000) Ionizing radiation and chemotherapeutic drugs induce apoptosis in lymphocytes in the absence of Fas or FADD/MORT1 signaling: implications for cancer therapy. J Exp Med 191:195

    Article  CAS  PubMed  Google Scholar 

  24. Nicoletti I, Migliorati G, Pagliacci MC, Grignani F, Riccardi C (1991) A rapid and simple method for measuring thymocyte apoptosis by propidium iodide staining and flow cytometry. J Immunol Methods 139:271

    Article  CAS  PubMed  Google Scholar 

  25. Nitobe J, Yamaguchi S, Okuyama M, Nozaki N, Sata M, Miyamoto T, Takeishi Y, Kubota I, Tomoike H (2003) Reactive oxygen species regulate FLICE inhibitory protein (FLIP) and susceptibility to Fas-mediated apoptosis in cardiac myocytes. Cardiovasc Res 57:119

    Article  CAS  PubMed  Google Scholar 

  26. Olson RD, Mushlin PS (1990) Doxorubicin cardiotoxicity: analysis of prevailing hypotheses. FASEB J 4:3076

    CAS  PubMed  Google Scholar 

  27. Pastorino F, Brignole C, Marimpietri D, Cilli M, Gambini C, Ribatti D, Longhi R, Allen TM, Corti A, Ponzoni M (2003) Vascular damage and anti-angiogenic effects of tumor vessel-targeted liposomal chemotherapy. Cancer Res 63:7400

    CAS  PubMed  Google Scholar 

  28. Petak I, Houghton JA (2001) Shared pathways: death receptors and cytotoxic drugs in cancer therapy. Pathol Oncol Res 7:95

    CAS  PubMed  Google Scholar 

  29. Scaffidi C, Fulda S, Srinivasan A, Friesen C, Li F, Tomaselli KJ, Debatin KM, Krammer PH, Peter ME (1998) Two CD95 (APO-1/Fas) signaling pathways. EMBO J 17:1675

    Article  CAS  PubMed  Google Scholar 

  30. Springhorn JP, Claycomb WC (1989) Preproenkephalin mRNA expression in developing rat heart and in cultured ventricular cardiac muscle cells. Biochem J 258:73

    CAS  PubMed  Google Scholar 

  31. Swain SM, Vici P (2004) The current and future role of dexrazoxane as a cardioprotectant in anthracycline treatment: expert panel review. J Cancer Res Clin Oncol 130:1

    Google Scholar 

  32. Um HD, Orenstein JM, Wahl SM (1996) Fas mediates apoptosis in human monocytes by a reactive oxygen intermediate dependent pathway. J Immunol 156:3469

    Google Scholar 

  33. Walczak H, Krammer PH (2000) The CD95 (APO-1/Fas) and the TRAIL (APO-2L) apoptosis systems. Exp Cell Res 256:58

    Article  CAS  PubMed  Google Scholar 

  34. Wallace KB (2003) Doxorubicin-induced cardiac mitochondrionopathy. Pharmacol Toxicol 93:105

    Article  CAS  PubMed  Google Scholar 

  35. Wang L, Ma W, Markovich R, Lee WL, Wang PH (1998) Insulin-like growth factor I modulates induction of apoptotic signaling in H9C2 cardiac muscle cells. Endocrinology 139:1354

    Article  CAS  PubMed  Google Scholar 

  36. Webb J (1963) Enzyme and metabolic inhibitors. Academic Press, New York

    Google Scholar 

  37. Wu S, Ko YS, Teng MS, Ko YL, Hsu LA, Hsueh C, Chou YY, Liew CC, Lee YS (2002) Adriamycin-induced cardiomyocyte and endothelial cell apoptosis: in vitro and in vivo studies. J Mol Cell Cardiol 34:1595

    Article  CAS  PubMed  Google Scholar 

  38. Yamaoka M, Yamaguchi S, Suzuki T, Okuyama M, Nitobe J, Nakamura N, Mitsui Y, Tomoike H (2000) Apoptosis in rat cardiac myocytes induced by Fas ligand: priming for Fas-mediated apoptosis with doxorubicin. J Mol Cell Cardiol 32:881

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

The authors thank Renata Zucic, Petra Berger, and Ursula Nägele for excellent technical help.

Author information

Authors and Affiliations

  1. Department of Hematology/Oncology, Dr. von Haunersches Kinderspital, Lindwurmstr 4, 80337, Munich, Germany

    Irmela Jeremias

  2. University Clinic and Policlinic for Children and Adolescents, Prittwitzstr 43, 89075, Ulm, Germany

    Karsten Stahnke & Klaus -Michael Debatin

Authors
  1. Irmela Jeremias
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Karsten Stahnke
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Klaus -Michael Debatin
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Klaus -Michael Debatin.

Additional information

This work was supported by Wilhelm Sander Stiftung and Bettina-Bräu-Stiftung.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Jeremias, I., Stahnke, K. & Debatin, K.M. CD95/Apo-1/Fas: independent cell death induced by doxorubicin in normal cultured cardiomyocytes. Cancer Immunol Immunother 54, 655–662 (2005). https://doi.org/10.1007/s00262-004-0604-y

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00262-004-0604-y

Keywords

Search

Navigation