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Cell cycle-specific metabolism of arabinosyl nucleosides in K562 human leukemia cells

Summary

Exponentially growing K562 cells incubated with 1-β-d-arabinofuranosylcytosine (ara-C) accumulate ara-C triphosphate (ara-CTP) at a higher rate and to a greater concentration after pretreatment with 9-β-d-ara-binofuranosyl-2-fluoroadenine (F-ara-A) than do cells treated with ara-C alone. Potentiation of ara-C metabolism is due in part to an indirect effect of F-ara-A triphosphate (F-ara-ATP)-mediated reduction in deoxynucleotide pools and consequent activation of deoxycytidine kinase. Because the levels of deoxynucleotide pools and the activity of deoxycytidine kinase are cell cycle-specific, we investigated the effect of cell cycle phases on the accumulation of ara-CTP and the influence of F-ara-A pretreatment on such accumulation. Exponentially growing K562 cells were fractionated into G1, S, and G2+M phase-enriched subpopulations (each enriched by >60%) by centrifugal elutriation. The rate of ara-CTP accumulation was 22, 25, and 14 μm/h and the rate of F-ara-ATP accumulation was 38, 47, and 33 μm/h in the G1, S, and G2+M subpopulations, respectively. The rate of elimination of arabinosyl triphosphates was similar among the different phases of the cell cycle. After pretreatment with F-ara-A, the rate of ara-CTP accumulation in the G1, S, and G2+M phase-enriched subpopulations was 43, 37, and 26 μm/h, indicating a 1.7-, 1.5-, and 1.9-fold increase, respectively. These results suggest that a combination of F-ara-A and ara-C may effectively potentiate ara-CTP accumulation in all phases of the cell cycle. This observation is consistent with the results of studies on the modulation of ara-C metabolism by F-ara-A in lymphocytes and leukemia blasts obtained from patients with chronic lymphocytic leukemia and acute myelogenous leukemia, respectively.

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References

  1. 1.

    Adams RLP, Berryman S, Thomson A (1971) Deoxynucleoside triphosphate pools in synchronized and drug-inhibited L-929 cells. Biochim Biophys Acta 240:455

  2. 2.

    Andreeff M (1986) Cell kinetics of leukemia. Semin Hematol 23:300

  3. 3.

    Arner ESJ, Flygar M, Bohman C, Wallstrom B, Eriksson S (1988) Deoxycytidine kinase is constitutively expressed in human lymphocytes: consequences for compartmentation effects, unscheduled DNA synthesis, and viral replication in resting cells. Exp Cell Res 178:335

  4. 4.

    Bello LJ (1974) Regulation of thymidine kinase in human cells. Exp Cell Res 89:263

  5. 5.

    Bray G, Bent TP (1972) Deoxynucleoside 5′-triphosphate pool fluctuations during the mammalian cell cycle. Biochim Biophys Acta 269:184

  6. 6.

    Brent TP (1971) Periodicity of DNA synthetic enzymes during the HeLa cell cycle. Cell Tissue Kinet 4:297

  7. 7.

    Brent TP, Butler JAV, Cranthorn AR (1965) Variations in phosphokinase activities during cell cycle in synchronous populations of HeLa cells. Nature 207:176

  8. 8.

    Brockman RW, Cheng Y-C, Schabel FM Jr, Montgomery JA (1980) Metabolism and chemotherapeutic activity of 1-β-d-arabinofuranosyl-2-fluoroadenine against murine leukemia L1210 and evidence for its phosphorylation by deoxycytidine kinase. Cancer Res 40: 3610

  9. 9.

    Chu MY, Fischer GA (1965) Comparative studies of leukemia cells sensitive and resistant to cytosine arabinoside. Biochem Pharmacol 14:333

  10. 10.

    Cohen A, Barankiewicz J, Lederman HM, Gelfand FW (1983) Purine and pyrimidine metabolism in human T lymphocytes. Regulation of deoxynucleotide metabolism. J Biol Chem 258:12334

  11. 11.

    Dow LW, Bell DE, Poulakos L, Fridland A (1980) Differences in metabolism and cytotoxicity between 9-β-d-arabinofuranosyl-adenine and 9-β-d-arabinofuranosyl-2-fluoroadenine in human leukemic lymphoblasts. Cancer Res 40:1405

  12. 12.

    Durham JP, Ives DH (1969) Deoxycytidine kinase: I. Distribution in normal and neoplastic tissues and interrelationships of deoxycytidine and 1-β-d-arabinofuranosylcytosine phosphorylation. Mol Pharmacol 5:358

  13. 13.

    Freireich EJ (1987) Arabinosylcytosine: 20-year update. J Clin Oncol 5:523

  14. 14.

    Gandhi V, Plunkett W (1988) Modulation of arabinosyl nucleoside metabolism by arabinosylnucleotides in human leukemia cells. Cancer Res 48:329

  15. 15.

    Gandhi V, Plunkett W (1989) Interaction of arabinosyl nucleotides in K562 human leukemia cells. Biochem Pharmacol 38:3531

  16. 16.

    Gandhi V, Plunkett W (1990) Modulatory activity of 2′,2′-difluorodeoxycytidine on the phosphorylation and cytotoxicity of arabinosyl nucleosides. Cancer Res 50:3675

  17. 17.

    Gandhi V, Danhauser L, Plunkett W (1987) Separation of 1-β-d-arabinofuranosylcytosine 5′-triphosphate and 9-β-d-arabinofuranosyl-2-fluoroadenine 5′-triphosphate in human leukemia cells by high-performance liquid chromatography. J Chromatogr 413:293

  18. 18.

    Gandhi V, Nowak B, Keating MJ, Plunkett W (1989) Modulation of arabinosylcytosine metabolism by arabinosyl-2-fluoroadenine in lymphocytes from patients with chronic lymphocytic leukemia: implications for combination therapy. Blood 74:2070

  19. 19.

    Gandhi V, Estey E, Kemena A, Keating MJ, Plunkett W (1991) Potentiation of arabinosylcytosine (ara-C) triphosphate (TP) metabolism in leukemia blasts by fludarabine monophosphate (MP) during therapy. Proc Am Soc Clin Oncol 10:95

  20. 20.

    Gandhi V, Kemena A, Keating MJ, Plunkett W (1992) Fludarabine infusion potentiates arabinosylcytosine metabolism in lymphocytes of patients with chronic lymphocytic leukemia. Cancer Res 52:897

  21. 21.

    Howard DK, Hay J, Melvin WT, Durham JP (1974) Changes in DNA and RNA synthesis and associated enzyme activities after the stimulation of serum-depleted BHK21/C13 cells by the addition of serum. Exp Cell Res 86:31

  22. 22.

    Keating MJ, McCredie KB, Bodey GP, Smith TL, Gehan EA, Freireich EJ (1982) Improved prospects for long-term survival in adults with acute myelogenous leukemia. JAMA 248:2481

  23. 23.

    Keating MJ, Kantarjian H, Talpaz M, Redman J, Koller C, Barlogie B, Velasquez W, Plunkett W, Freireich EJ, McCredie KB (1989) Fludarabine: a new agent with major activity aganist chronic lymphocytic leukemia. Blood 74:19

  24. 24.

    Keating MJ, Kantarjian H, O'Brien S, Koller C, Talpaz M, Schachner J, Childs CC, Freireich EJ, McCredie KB (1991) Fludarabine: a new agent with marked cytoreductive activity in untreated chronic lymphocytic leukemia. J Clin Oncol 9:44

  25. 25.

    Liliemark JO, Plunkett W (1986) Regulation of 1-β-d-arabinofuranosylcytosine 5′-triphosphate accumulation in human leukemia cells by deoxycytidine 5′-triphosphate. Cancer Res 46:1079

  26. 26.

    Lozzio GB, Lozzio BB (1975) Human chronic myelogenous leukemia cell line with positive Philadelphia chromosome. Blood 45:321

  27. 27.

    Mathews CK, Slabaugh MB (1986) Eukaryotic DNA metabolism. Are DNA precursors channeled to replication sites? Exp Cell Res 162:285

  28. 28.

    Meistrich ML (1983) Experimental factors involved in separation by centrifugal elutriation. In: Pretlow TG III, Pretlow TP (eds) Cell separation: methods and selected applications, vol 2. Academic Press, New York, pp 33–61

  29. 29.

    Momparler RL, Fischer GA (1968) Mammalian deoxynucleoside kinase: I. Deoxycytidine kinase: purification, properties, and kinetic studies with cytosine arabinoside. J Biol Chem 243:4298

  30. 30.

    Neu HC, Heppel LA (1964) Nucleotide sequence analysis of polyri-bonucleotides by means of periodate oxidation followed by cleavage with an amine. J Biol Chem 239:2927

  31. 31.

    Novotný L, Plunkett W (1989) Synthesis of 9-β-d-arabinofuranosyl-2-fluoroadenine 5′-triphosphate. In: Townsend LB, and Tipson RS (eds) Nucleic acid chemistry, part 4. John Wiley & Sons, New York, pp 327–331

  32. 32.

    Plunkett W, Hug V, Keating MJ, Chubb S (1980) Quantitation of 1-β-d-arabinosylcytosine 5′-triphosphate in leukemic cells from bone marrow and peripheral blood of patients receiving 1-β-d-arabinofuranosylcytosine therapy. Cancer Res 40:588

  33. 33.

    Plunkett W, Chubb S, Alexander L, Montgomery JA (1980) Comparison of the toxicity and metabolism of 1-β-d-arabinofuranosyl-2-fluoroadenine and 1-β-d-arabinofuranosyladenine in human lymphoblastoid cells. Cancer Res 40:2349

  34. 34.

    Plunkett W, Liliemark JO, Estey E, Keating MJ (1987) Saturation of ara-CTP accumulation during high-dose ara-C therapy: pharmacologic rationale for intermediate dose ara-C. Semin Oncol 14 [Suppl 1]:159

  35. 35.

    Plunkett W, Liliemark JO, Adams TM, Nowak B, Estey E, Kantarjian H, Keating MJ (1987) Saturation of 1-β-d-arabinofuranosylcytosine 5′-triphosphate accumulation in leukemia cells during high-dose 1-β-d-arabinofuranosylcytosine therapy. Cancer Res 47:3005

  36. 36.

    Reichard P (1988) Interactions between deoxynucleotide and DNA synthesis. Annu Rev Biochem 57:349

  37. 37.

    Richel DJ, Colly LP, Arentsen-Honders MW, Starrenburg CWJ, Willemze R (1990) Deoxycytidine kinase, thymidine kinase, and cytidine deaminase and the formation of ara-CTP in leukemic cells in different phases of the cell cycle. Leukemia Res 14:363

  38. 38.

    Schrecker AW, Urshel MJ (1968) Metabolism of 1-β-d-arabinofuranosylcytosine in leukemia L1210: studies with intact cells. Cancer Res 28:793

  39. 39.

    Sherley JL, Kelly TJ (1988) Regulation of human thymidine kinase during the cell cycle. J Biol Chem 263:8350

  40. 40.

    Skoog L, Bjursell G (1974) Nuclear and cytoplasmic pool of deoxynucleoside triphosphates in Chinese hamster ovary cells. J Biol Chem 249:6434

  41. 41.

    Tseng W-C, Derse D, Cheng Y-C, Brockman RW, Bennett LL Jr (1982) In vitro biological activity of 9-β-d-arabinofuranosyl-2-fluoroadenine and the biochemical actions of its triphosphate on DNA polymerases and ribonucleotide reductase from HeLa cells. Mol Pharmacol 21:474

  42. 42.

    Vierwinden G, Drenthe-Schonk AM, Plas AM, Linssen PCM, Pennings AHM, Holdrinet RSG, Egmond JV, Wessel JMC, Haanen CAM (1982) Variations of the phosphorylation of 1-β-arabinofuranosylcytosine (ara-C) in human myeloid leukemic cells related to the cell cycle. Leukemia Res 6:251

  43. 43.

    Wan CW, Mak TW (1978) Deoxycytidine kinase and cytosine nucleoside deaminase activities in synchronized cultures of normal rat kidney cells. Cancer Res 38:1768

  44. 44.

    White EL, Shaddix SC, Brockman RW, Bennett LL Jr (1982) Comparison of the actions of 9-β-d-arabinofuranosyl-2-fluoroadenine and 9-β-d-arabinofuranosyladenine on target enzymes from mouse tumor cells. Cancer Res 42:2260

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Author information

Correspondence to Varsha Gandhi.

Additional information

Supported in part by grants CA 28596 and CA 16672 from the National Cancer Institute, Department of Health and Human Services, and by grant DHP-I from the American Cancer Society

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Gandhi, V., Plunkett, W. Cell cycle-specific metabolism of arabinosyl nucleosides in K562 human leukemia cells. Cancer Chemother. Pharmacol. 31, 11–17 (1992). https://doi.org/10.1007/BF00695988

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Keywords

  • Leukemia
  • Nucleoside
  • Chronic Lymphocytic Leukemia
  • Acute Myelogenous Leukemia
  • Cell Cycle Phase