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. 1999 Mar;79(7-8):1090–1097. doi: 10.1038/sj.bjc.6690174

Dual mechanism of daunorubicin-induced cell death in both sensitive and MDR-resistant HL-60 cells

M-G Côme 1,2, A Skladanowski 2,3, A K Larsen 2, G Laurent 1,4
PMCID: PMC2362243  PMID: 10098741

Abstract

Exposure of some acute myeloid leukaemia (AML) cells to daunorubicin leads to rapid cell death, whereas other AML cells show natural drug resistance. This has been attributed to expression of functional P-glycoprotein resulting in reduced drug accumulation. However, it has also been proposed that P-glycoprotein-expressing multidrug-resistant (MDR) cells are inherently defective for apoptosis. To distinguish between these different possibilities, we have compared the cell death process in a human AML cell line (HL-60) with a MDR subline (HL-60/Vinc) at doses that yield either similar intracellular daunorubicin concentrations or comparable cytotoxicity. Adjustment of the dose to obtain the same intracellular drug accumulation in the two cell lines did not result in equal cytotoxicity, suggesting the presence of additional resistance mechanisms in the P-glycoprotein-expressing HL-60/Vinc cells. However, at equitoxic doses, similar cell death pathways were observed. In HL-60 cells, daunorubicin induced rapid apoptosis at 0.5–1 μM and delayed mitotic cell death at 0.1 μM. These concentrations are within the clinical dose range. Similarly, HL-60/Vinc cells underwent apoptosis at 50–100 μM daunorubicin and mitotic cell death at 10 μM. These results show, for the first time, that anthracyclines can induce cell death by a dual mechanism in both sensitive and MDR cells. Our results also show that not only the cytotoxicity, but also the kinetics and mechanism of cell death, are dose dependent. Interestingly, regrowth was observed only in association with delayed cell death and the formation of enlarged, often polyploid, cells with micronucleation, suggesting that morphological criteria may be useful to evaluate treatment efficacy in patients with myeloid leukaemias. © 1999 Cancer Research Campaign

Keywords: drug resistance, P-glycoprotein, cell death, daunorubicin, myeloid leukaemia, response prediction

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Selected References

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  1. Agarwal M. L., Clay M. E., Harvey E. J., Evans H. H., Antunez A. R., Oleinick N. L. Photodynamic therapy induces rapid cell death by apoptosis in L5178Y mouse lymphoma cells. Cancer Res. 1991 Nov 1;51(21):5993–5996. [PubMed] [Google Scholar]
  2. Baer M. R., Bloomfield C. D. Multidrug resistance in acute myeloid leukemia. J Natl Cancer Inst. 1991 May 15;83(10):663–665. doi: 10.1093/jnci/83.10.663. [DOI] [PubMed] [Google Scholar]
  3. Bhuyan B. K., Groppi V. E. Cell cycle specific inhibitors. Pharmacol Ther. 1989;42(3):307–348. doi: 10.1016/0163-7258(89)90029-6. [DOI] [PubMed] [Google Scholar]
  4. Chang W. P., Little J. B. Delayed reproductive death as a dominant phenotype in cell clones surviving X-irradiation. Carcinogenesis. 1992 Jun;13(6):923–928. doi: 10.1093/carcin/13.6.923. [DOI] [PubMed] [Google Scholar]
  5. Cucco C., Calabretta B. In vitro and in vivo reversal of multidrug resistance in a human leukemia-resistant cell line by mdr1 antisense oligodeoxynucleotides. Cancer Res. 1996 Oct 1;56(19):4332–4337. [PubMed] [Google Scholar]
  6. Darzynkiewicz Z., Carter S. P., Mikulski S. M., Ardelt W. J., Shogen K. Cytostatic and cytotoxic effects of Pannon (P-30 Protein), a novel anticancer agent. Cell Tissue Kinet. 1988 May;21(3):169–182. doi: 10.1111/j.1365-2184.1988.tb00855.x. [DOI] [PubMed] [Google Scholar]
  7. Demarcq C., Bunch R. T., Creswell D., Eastman A. The role of cell cycle progression in cisplatin-induced apoptosis in Chinese hamster ovary cells. Cell Growth Differ. 1994 Sep;5(9):983–993. [PubMed] [Google Scholar]
  8. Eguchi Y., Shimizu S., Tsujimoto Y. Intracellular ATP levels determine cell death fate by apoptosis or necrosis. Cancer Res. 1997 May 15;57(10):1835–1840. [PubMed] [Google Scholar]
  9. Frankfurt O. S., Seckinger D., Sugarbaker E. V. Pleiotropic drug resistance and survival advantage in leukemic cells with diminished apoptotic response. Int J Cancer. 1994 Oct 15;59(2):217–224. doi: 10.1002/ijc.2910590214. [DOI] [PubMed] [Google Scholar]
  10. Gervasoni J. E., Jr, Fields S. Z., Krishna S., Baker M. A., Rosado M., Thuraisamy K., Hindenburg A. A., Taub R. N. Subcellular distribution of daunorubicin in P-glycoprotein-positive and -negative drug-resistant cell lines using laser-assisted confocal microscopy. Cancer Res. 1991 Sep 15;51(18):4955–4963. [PubMed] [Google Scholar]
  11. Gorczyca W., Gong J., Darzynkiewicz Z. Detection of DNA strand breaks in individual apoptotic cells by the in situ terminal deoxynucleotidyl transferase and nick translation assays. Cancer Res. 1993 Apr 15;53(8):1945–1951. [PubMed] [Google Scholar]
  12. Gottesman M. M., Pastan I. Biochemistry of multidrug resistance mediated by the multidrug transporter. Annu Rev Biochem. 1993;62:385–427. doi: 10.1146/annurev.bi.62.070193.002125. [DOI] [PubMed] [Google Scholar]
  13. Gromova I. I., Thomsen B., Razin S. V. Different topoisomerase II antitumor drugs direct similar specific long-range fragmentation of an amplified c-MYC gene locus in living cells and in high-salt-extracted nuclei. Proc Natl Acad Sci U S A. 1995 Jan 3;92(1):102–106. doi: 10.1073/pnas.92.1.102. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Hickman J. A. Apoptosis induced by anticancer drugs. Cancer Metastasis Rev. 1992 Sep;11(2):121–139. doi: 10.1007/BF00048059. [DOI] [PubMed] [Google Scholar]
  15. Hindenburg A. A., Gervasoni J. E., Jr, Krishna S., Stewart V. J., Rosado M., Lutzky J., Bhalla K., Baker M. A., Taub R. N. Intracellular distribution and pharmacokinetics of daunorubicin in anthracycline-sensitive and -resistant HL-60 cells. Cancer Res. 1989 Aug 15;49(16):4607–4614. [PubMed] [Google Scholar]
  16. Keizer H. G., Schuurhuis G. J., Broxterman H. J., Lankelma J., Schoonen W. G., van Rijn J., Pinedo H. M., Joenje H. Correlation of multidrug resistance with decreased drug accumulation, altered subcellular drug distribution, and increased P-glycoprotein expression in cultured SW-1573 human lung tumor cells. Cancer Res. 1989 Jun 1;49(11):2988–2993. [PubMed] [Google Scholar]
  17. Ling Y. H., Priebe W., Perez-Soler R. Apoptosis induced by anthracycline antibiotics in P388 parent and multidrug-resistant cells. Cancer Res. 1993 Apr 15;53(8):1845–1852. [PubMed] [Google Scholar]
  18. Ling Y. H., el-Naggar A. K., Priebe W., Perez-Soler R. Cell cycle-dependent cytotoxicity, G2/M phase arrest, and disruption of p34cdc2/cyclin B1 activity induced by doxorubicin in synchronized P388 cells. Mol Pharmacol. 1996 May;49(5):832–841. [PubMed] [Google Scholar]
  19. Lock R. B., Galperina O. V., Feldhoff R. C., Rhodes L. J. Concentration-dependent differences in the mechanisms by which caffeine potentiates etoposide cytotoxicity in HeLa cells. Cancer Res. 1994 Sep 15;54(18):4933–4939. [PubMed] [Google Scholar]
  20. Lock R. B., Ross W. E. Possible role for p34cdc2 kinase in etoposide-induced cell death of Chinese hamster ovary cells. Cancer Res. 1990 Jun 15;50(12):3767–3771. [PubMed] [Google Scholar]
  21. Lock R. B., Stribinskiene L. Dual modes of death induced by etoposide in human epithelial tumor cells allow Bcl-2 to inhibit apoptosis without affecting clonogenic survival. Cancer Res. 1996 Sep 1;56(17):4006–4012. [PubMed] [Google Scholar]
  22. Marie J. P., Zhou D. C., Gurbuxani S., Legrand O., Zittoun R. MDR1/P-glycoprotein in haematological neoplasms. Eur J Cancer. 1996 Jun;32A(6):1034–1038. doi: 10.1016/0959-8049(96)00055-x. [DOI] [PubMed] [Google Scholar]
  23. McGrath T., Latoud C., Arnold S. T., Safa A. R., Felsted R. L., Center M. S. Mechanisms of multidrug resistance in HL60 cells. Analysis of resistance associated membrane proteins and levels of mdr gene expression. Biochem Pharmacol. 1989 Oct 15;38(20):3611–3619. doi: 10.1016/0006-2952(89)90134-2. [DOI] [PubMed] [Google Scholar]
  24. Meschini S., Molinari A., Calcabrini A., Citro G., Arancia G. Intracellular localization of the antitumour drug adriamycin in living cultured cells: a confocal microscopy study. J Microsc. 1994 Dec;176(Pt 3):204–210. doi: 10.1111/j.1365-2818.1994.tb03516.x. [DOI] [PubMed] [Google Scholar]
  25. Quillet-Mary A., Mansat V., Duchayne E., Come M. G., Allouche M., Bailly J. D., Bordier C., Laurent G. Daunorubicin-induced internucleosomal DNA fragmentation in acute myeloid cell lines. Leukemia. 1996 Mar;10(3):417–425. [PubMed] [Google Scholar]
  26. Rodilla V. Origin and evolution of binucleated cells and binucleated cells with micronuclei in cisplatin-treated CHO cultures. Mutat Res. 1993 Aug;300(3-4):281–291. doi: 10.1016/0165-1218(93)90062-i. [DOI] [PubMed] [Google Scholar]
  27. Skladanowski A., Konopa J. Adriamycin and daunomycin induce programmed cell death (apoptosis) in tumour cells. Biochem Pharmacol. 1993 Aug 3;46(3):375–382. doi: 10.1016/0006-2952(93)90512-u. [DOI] [PubMed] [Google Scholar]
  28. Speth P. A., Linssen P. C., Boezeman J. B., Wessels H. M., Haanen C. Leukemic cell and plasma daunomycin concentrations after bolus injection and 72 h infusion. Cancer Chemother Pharmacol. 1987;20(4):311–315. doi: 10.1007/BF00262582. [DOI] [PubMed] [Google Scholar]
  29. Sugimoto K., Toyoshima H., Sakai R., Miyagawa K., Hagiwara K., Ishikawa F., Takaku F., Yazaki Y., Hirai H. Frequent mutations in the p53 gene in human myeloid leukemia cell lines. Blood. 1992 May 1;79(9):2378–2383. [PubMed] [Google Scholar]
  30. Sun X. M., Snowden R. T., Dinsdale D., Ormerod M. G., Cohen G. M. Changes in nuclear chromatin precede internucleosomal DNA cleavage in the induction of apoptosis by etoposide. Biochem Pharmacol. 1994 Jan 20;47(2):187–195. doi: 10.1016/0006-2952(94)90005-1. [DOI] [PubMed] [Google Scholar]
  31. Taatjes D. J., Gaudiano G., Resing K., Koch T. H. Redox pathway leading to the alkylation of DNA by the anthracycline, antitumor drugs adriamycin and daunomycin. J Med Chem. 1997 Apr 11;40(8):1276–1286. doi: 10.1021/jm960835d. [DOI] [PubMed] [Google Scholar]
  32. Thompson C. B. Apoptosis in the pathogenesis and treatment of disease. Science. 1995 Mar 10;267(5203):1456–1462. doi: 10.1126/science.7878464. [DOI] [PubMed] [Google Scholar]
  33. Tobey R. A. Effects of cytosine arabinoside, daunomycin, mithramycin, azacytidine, adriamycin, and camptothecin on mammalian cell cycle traverse. Cancer Res. 1972 Dec;32(12):2720–2725. [PubMed] [Google Scholar]
  34. Tounekti O., Pron G., Belehradek J., Jr, Mir L. M. Bleomycin, an apoptosis-mimetic drug that induces two types of cell death depending on the number of molecules internalized. Cancer Res. 1993 Nov 15;53(22):5462–5469. [PubMed] [Google Scholar]
  35. Tsao Y. P., D'Arpa P., Liu L. F. The involvement of active DNA synthesis in camptothecin-induced G2 arrest: altered regulation of p34cdc2/cyclin B. Cancer Res. 1992 Apr 1;52(7):1823–1829. [PubMed] [Google Scholar]
  36. Weinert T. A. Dual cell cycle checkpoints sensitive to chromosome replication and DNA damage in the budding yeast Saccharomyces cerevisiae. Radiat Res. 1992 Nov;132(2):141–143. [PubMed] [Google Scholar]
  37. Weinert T. A., Hartwell L. H. The RAD9 gene controls the cell cycle response to DNA damage in Saccharomyces cerevisiae. Science. 1988 Jul 15;241(4863):317–322. doi: 10.1126/science.3291120. [DOI] [PubMed] [Google Scholar]

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