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Sensitivity, Resistance to Therapy and Apoptosis

Study of apoptosis-related responses of leukemic blast cells to in vitro anthracycline treatment

Leukemia volume 14, pages 1266–1275 (2000)Cite this article

Abstract

Anthracyclines trigger an apoptotic cell death but their molecular targets are not totally explored. We investigated the apoptotic response of blast cells and lymphocytes from medullary samples of 31 de novo acute leukemia. Mononuclear cells were treated in vitroby therapeutic concentrations of either daunorubicin (DNR) or idarubicin (IDA) for 1 h, washed and cultured for 18 h. A multivariate analysis using flow cytometry and a CD45 gating on lymphocytes and blast cells was performed. DNR and IDA induced a Fas enhancement on both leukemic and normal cells. In blast cells the DEVDases were activated and the caspase 3 was cleaved in relation to phosphatidyl serine exposure, showing a caspase-dependent pathway in anthracycline-induced apoptosis. Apoptotic percentages were always higher for blast cells than for lymphocytes, confirming that anthracycline toxicity mainly affected tumor cells. Moreover, drug-induced apoptosis was not related to spontaneous apoptosis, suggesting that variations in response intensities were due to individual variations of sensitivity rather than to programmed life span time. The apoptotic response of P- glycoprotein-expressing blast cells was not significant, giving biological argument for the poor prognosis of multidrug resistance leukemia. Finally, Fas induction and anthracycline-induced apoptosis on blast cells were significantly higher when a complete remission was achieved, thus shedding light on potential new prognostic factors in acute leukemia.

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References

  1. Nagata S . Apoptosis by death factor Cell 1997 88: 355–365

    Article  CAS  Google Scholar 

  2. 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

    Article  CAS  Google Scholar 

  3. Boldin MP, Varfolomeev EE, Pancer Z, Mett IL, Camonis JH, Wallach D . A novel protein that interacts with the death domain of Fas/APO1 contains a sequence motif related to the death domain J Biol Chem 1995 270: 7795–7798

    Article  CAS  Google Scholar 

  4. 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

    Article  CAS  Google Scholar 

  5. Muzzio M, Chinnaiyan AM, Kischkel FC, O'Rourke K, Shevchenko A, Ni J, Scaffidi C, Bretz JD, Zhang M, Gentz R, Mann M, Krammer PH, Peter ME, Dixit VM . 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

    Article  Google Scholar 

  6. Medema JP, Scaffidi C, Kischkel FC, Shevchenko A, Mann M, Krammer PH, Peter ME . FLICE is activated by association with the Cd95 death-inducing signaling complex (DISC) EMBO J 1997 16: 2794–2804

    Article  CAS  Google Scholar 

  7. Thornberry NA, Lazebnik Y . Caspases: enemies within Science 1998 281: 1312–1316

    Article  CAS  Google Scholar 

  8. Solary E, Micheau O, Dimanche-Boitrel MT, Martin F . Role of the Fas/Fas-L system in the immune response to tumors and the resistance to cytotoxic drugs Bull Cancer 1998 85: 685–694

    CAS  PubMed  Google Scholar 

  9. Dirks W, Schöne S, Uphoff C, Quentmeier H, Pradella S, Drexler HG . Expression and function of CD95 (Fas/APO-1) in leukemia–lymphoma tumour lines Br J Haematol 1997 96: 584–593

    Article  CAS  Google Scholar 

  10. Keane MM, Ettenberg SA, Lowrey GA, Russell EK, Lipkowirz S . Fas expression and function in normal and malignant breast cell lines Cancer Res 1996 56: 4791–4798

    CAS  PubMed  Google Scholar 

  11. Landowski TH, Gleason-Guzman MC, Dalton WS . Selection for drug resistance to Fas-mediated apoptosis Blood 1997 89: 1854–1861

    CAS  PubMed  Google Scholar 

  12. Eischen CM, Kottke TJ, Martins LM, Basi GS, Tung JS, Earnshaw WC, Leibson PJ, Kaufmann SH . Comparison of apoptosis in wild-type and Fas-resistant cells: chemotherapy-induced apoptosis is not dependent on Fas/Fas-ligand interactions Blood 1997 90: 935–943

    CAS  PubMed  Google Scholar 

  13. Mizutani Y, Okada Y, Yoshida O, Fukumoto M, Bonavida B . Doxorubicin sensitizes human bladder carcinoma cells to Fas-mediated cytotoxicity Cancer 1997 79: 1180–1189

    Article  CAS  Google Scholar 

  14. Morimoto H, Yonehara S, Bonavida B . Overcoming tumor necrosis factor and drug resistance of human cell lines by combination treatment with anti-Fas antibody and drugs or toxins Cancer Res 1993 53: 2591–2596

    CAS  PubMed  Google Scholar 

  15. Nakamura S, Takeshima M, Nakamura Y, Ohtake S, Matsuda T . Induction of apoptosis in HL60 leukemic cells by anticancer drugs in combination with anti-Fas monoclonal antibody Anticancer Res 1997 17: 173–179

    CAS  PubMed  Google Scholar 

  16. Uslu R, Jewett A, Bonavida B . Sensitization of human ovarian tumor cells by subcytotoxic CDDP to anti-Fas antibody-mediated cytotoxicity and apoptosis Gynecol Oncol 1996 62: 232–291

    Article  Google Scholar 

  17. Micheau O, Solary E, Hammann A, Martin F, Dimanche-Boitrel MT . Sensitization of cancer cells treated with cytotoxic drugs to Fas-mediated cytotoxicity J Natl Cancer Inst 1997 89: 783–789

    Article  CAS  Google Scholar 

  18. Kimura H, Yamaguchi Y . A phase III randomized study of interleukin-2 lymphokine-activated killer cell immunotherapy combined with chemotherapy or radiotherapy after curative or noncurative resection of primary lung carcinoma Cancer 1997 80: 42–49

    Article  CAS  Google Scholar 

  19. Ling YH, Priebe W, Perez-Soler R . Apoptosis induced by anthracycline antibiotics in P388 parent and multidrug-resistant cells Cancer Res 1993 53: 1845–1852

    CAS  PubMed  Google Scholar 

  20. Skladanowski A, Konopa J . Adriamycin and daunomycin induce programmed cell death (apoptosis) in tumour cells Biochem Pharmacol 1993 46: 375–382

    Article  CAS  Google Scholar 

  21. Zaleskis G, Berleth E, Verstovsek S, Ehrke MJ, Mihich E . Doxorubicin-induced DNA degradation in murine thymocytes Mol Pharmacol 1994 46: 901–908

    CAS  PubMed  Google Scholar 

  22. Gewirtz DA . A critical evaluation of the mechanisms of action proposed for the antitumor effects of the anthracycline antibiotics adriamycin and daunorubicin Biochem Pharmacol 1999 57: 727–741

    Article  CAS  Google Scholar 

  23. Vial JP, Belloc F, Dumain P, Besnard S, Lacombe F, Boisseau MR, Reiffers J, Bernard P . Study of the apoptosis induced in vitro by antitumoral drugs on leukemic cells Leukemia Res 1997 21: 163–172

    Article  CAS  Google Scholar 

  24. Durrieu F, Belloc F, Lacoste L, Dumain P, Chabrol J, Dachary-Prigent J, Morjani H, Boisseau MR, Reiffers J, Bernard P, Lacombe F . Caspase activation is an early event in anthracycline-induced apoptosis and allows detection of apoptotic cells before they are ingested by phagocytes Exp Cell Res 1998 240: 165–175

    Article  CAS  Google Scholar 

  25. Friesen C, Fulda S, Debatin KM . Deficient activation of the CD95 (APO-l/Fas) system in drug-resistant cells Leukemia 1997 11: 1833–1841

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  27. Müller M, Strand S, Hug H, Heinemann EM, Walczak H, Hofmann WJ, Stremmel W, Krammer PH, Galle PR . 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

    Article  Google Scholar 

  28. Droin N, Dubrez L, Eymin B, Renvoizé C, Breard J, Dimanche-Boitrel MT, Solary E . Upregulation CASP genes in human tumor cells undergoing etoposide-induced apoptosis Oncogene 1998 16: 2885–2894

    Article  CAS  Google Scholar 

  29. Yu DK . The contribution of P-glycoprotein to pharmacokinetic drug–drug interactions J Clin Pharmacol 1999 39: 1203–1211

    Article  CAS  Google Scholar 

  30. Praet M, Stryckmans P, Ruysschaert JM . Cellular uptake, and transport kinetics of anthracyclines in human sensitive and multidrug-resistant K562 cells Biochem Pharmacol 1996 51: 1341–1348

    Article  CAS  Google Scholar 

  31. Gottesman MM, Pastan I . Biochemistry of multidrug resistance mediated by the multidrug transporter Ann Rev Biochem 1993 62: 385–425

    Article  CAS  Google Scholar 

  32. Frankfurt OS, Seckinger D, Sugarbaker EV . Pleiotropic drug resistance and survival advantage in leukemia cells with diminished apoptotic response Int J Cancer 1994 59: 217–224

    Article  CAS  Google Scholar 

  33. Bennett JM, Catovsky D, Daniel MT, Flandrin G, Galton DAG, Gralnick HR, Sultan C . Proposal for the classification of the acute leukaemias Br J Haematol 1976 33: 451–458

    Article  CAS  Google Scholar 

  34. Lacombe F, Durrieu F, Briais A, Dumain P, Belloc F, Bascans E, Reiffers J, Boisseau MR, Bernard P . Flow cytometry CD45 gating for immunophenotyping of acute myeloid leukemia Leukemia 1997 11: 1878–1886

    Article  CAS  Google Scholar 

  35. Belaud-Rotureau MA, Lacombe F, Durrieu F, Vial JP, Lacoste C, Bernard P, Belloc F . Ceramide-induced apoptosis occurs independently of caspases and is decreased by leupeptin Cell Death Differ 1999 6: 788–795

    Article  CAS  Google Scholar 

  36. Kuwazuru Y, Yoshimura A, Hanada S, Utsunomiya A, Makino T, Ishibashi K, Kodama M, Iwahashi M, Arima T, Akiyama S . Expression of the multidrug transporter, P-glycoprotein, in acute leukemia cells and correlation to clinical drug resistance Cancer 1990 66: 868–873

    Article  CAS  Google Scholar 

  37. Martin SJ, Green DR . Protease activation during apoptosis: death by a thousand cuts? Cell 1995 82: 349–352

    Article  CAS  Google Scholar 

  38. Green DR . Apoptotic pathways: the roads to ruin Cell 1998 94: 695–698

    Article  CAS  Google Scholar 

  39. Zubrod CG . Treatment of the acute leukemias Cancer Res 1967 27: 2557–2560

    CAS  PubMed  Google Scholar 

  40. Robert J, Gianni L . Pharmacokinetics and metabolism of anthracyclines Cancer Surv 1993 17: 219–251

    CAS  PubMed  Google Scholar 

  41. Borchmann P, Hubel K, Schnell R, Engert A . Idarubicin: a brief overview on pharmacology and clinical use Int J Clin Pharmacol Ther 1997 35: 80–83

    CAS  PubMed  Google Scholar 

  42. Campos L, Guyotat D, Archimbaud E, Calmard-Oriol P, Tsuruo T, Troncy J, Treille D, Fiere D . Clinical significance of multidrug resistance P-glycoprotein expression on acute nonlymphoblastic leukemia cells at diagnosis Blood 1992 79: 473–476

    CAS  PubMed  Google Scholar 

  43. Leith CP, Kopecky KJ, Chen IM, Eijdems L, Slovak ML, McConnell TS, Head DR, Weick J, Grever MR, Appelbaum FR, Willman CL . Frequency and clinical significance of the expression of the multidrug resistance proteins MDR1/P-glycoprotein, MRP1, and LRP in acute myeloid leukemia. A Southwest Oncology Group study Blood 1999 94: 1086–1099

    CAS  PubMed  Google Scholar 

  44. Coon JS, Wang Y, Bines SD, Markham PN, Chong ASF, Gebel HM . Multidrug resistance activity in human lymphocytes Hum Immunol 1991 32: 134–140

    Article  CAS  Google Scholar 

  45. Drach D, Zhao S, Drach J, Mahadevia R, Gattringer C, Huber H, Andreeff M . Subpopulations of normal peripheral blood and bone marrow cells express a functional multidrug resistance phenotype Blood 1992 80: 2729–2734

    CAS  PubMed  Google Scholar 

  46. Klimeckj WT, Furscher BW, Grogan TM, Dalton WS . P-glycoprotein expression and function in circulating blood cells from normal volunteers Blood 1994 83: 2451–2458

    Google Scholar 

  47. Drach J, Gsur A, Hamilton G, Zhao S, Angerler J, Fiegl M, Zojer N, Raderer M, Haberl I, Andreeff M, Huber H . Involvement of P-glycoprotein in the transmembrane transport of interleukin-2 (IL-2), IL-4, and interferon-gamma in normal human T lymphocytes Blood 1996 88: 1747–1754

    CAS  PubMed  Google Scholar 

  48. Raymond M, Gros P, Whiteway M, Thomas DY . Functional complementation of yeast ste6 by mammalian multidrug resistance mdr gene Science 1992 256: 232–234

    Article  CAS  Google Scholar 

  49. Durrieu F, Belaud-Rotureau MA, Lacombe F, Dumain P, Reiffers J, Boisseau MR, Bernard P, Belloc F . Synthesis of Bcl-2 in response to anthracycline treatment may contribute to an apoptosis-resistant phenotype in leukemic cell lines Cytometry 1999 36: 140–149

    Article  CAS  Google Scholar 

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Acknowledgements

This work was supported in part by grants from the Ligue Nationale contre le Cancer and from the Association pour la Recherche sur le Cancer.

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Authors and Affiliations

  1. Laboratoire d'Hématologie, Hôpital du Haut-Lévêque, Pessac, France

    M-A Belaud-Rotureau, F Durrieu, F Lacombe & F Belloc

  2. Laboratoire Universitaire d'Hématologie, Université Victor Segalen, Bordeaux, France

    M-A Belaud-Rotureau, G Labroille, G Marit & F Belloc

  3. CNRS-UMR 5540, Université Victor Segalen, Bordeaux, France

    F Lacombe & J Reiffers

  4. Service des Maladies du Sang, Hôpital du Haut-Lévêque, Pessac, France

    O Fitoussi, P Agape, G Marit & J Reiffers

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Belaud-Rotureau, MA., Durrieu, F., Labroille, G. et al. Study of apoptosis-related responses of leukemic blast cells to in vitro anthracycline treatment. Leukemia 14, 1266–1275 (2000). https://doi.org/10.1038/sj.leu.2401803

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