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. 1987 Feb;79(2):380–387. doi: 10.1172/JCI112823

Membrane transport influences the rate of accumulation of cytosine arabinoside in human leukemia cells.

J C White, J P Rathmell, R L Capizzi
PMCID: PMC424079  PMID: 3805274

Abstract

The role of membrane transport in the cellular accumulation of 1-beta-D-arabinofuranosylcytosine (ara-C) was studied in freshly isolated human acute leukemia cells. Patient cells had low rates for ara-C transport as compared with human and murine experimental cells and correspondingly low binding capacities for the nucleoside transport inhibitor, nitrobenzylmercaptopurine riboside (NBMPR). At 1 microM ara-C, the rate of net cellular accumulation was close to the membrane transport rate, and NBMPR inhibited transport and accumulation to the same extent. The rate of ara-C accumulation was half maximal at only 3-5 microM, a level much lower than that required for murine cells (67-85 microM). At concentrations below 1 microM the rate of ara-C accumulation was determined primarily by the transport rate, but at higher concentrations above 10 microM, phosphorylation capacity was the principal determinant of the net uptake rate. This difference in the role of transport at high and low ara-C concentrations may explain, in part, the efficacy of high-dose ara-C in patients refractory to standard dose protocols.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bach M. K. Biochemical and genetic studies of a mutant strain of mouse leukemia L1210 resistant to 1-beta-D-arabinofuranosylcytosine (Cytarabine). Cancer Res. 1969 May;29(5):1036–1044. [PubMed] [Google Scholar]
  2. Belt J. A. Heterogeneity of nucleoside transport in mammalian cells. Two types of transport activity in L1210 and other cultured neoplastic cells. Mol Pharmacol. 1983 Nov;24(3):479–484. [PubMed] [Google Scholar]
  3. Bhalla K., Nayak R., Grant S. Isolation and characterization of a deoxycytidine kinase-deficient human promyelocytic leukemic cell line highly resistant to 1-beta-D- arabinofuranosylcytosine. Cancer Res. 1984 Nov;44(11):5029–5037. [PubMed] [Google Scholar]
  4. Bowen D., Diasio R. B., Goldman I. D. Distinguishing between membrane transport and intracellular metabolism of fluorodeoxyuridine in Ehrlich ascites tumor cells by application of kinetic and high performance liquid chromatographic techniques. J Biol Chem. 1979 Jun 25;254(12):5333–5339. [PubMed] [Google Scholar]
  5. Capizzi R. L., Poole M., Cooper M. R., Richards F., 2nd, Stuart J. J., Jackson D. V., Jr, White D. R., Spurr C. L., Hopkins J. O., Muss H. B. Treatment of poor risk acute leukemia with sequential high-dose ARA-C and asparaginase. Blood. 1984 Mar;63(3):694–700. [PubMed] [Google Scholar]
  6. Capizzi R. L., Yang J. L., Cheng E., Bjornsson T., Sahasrabudhe D., Tan R. S., Cheng Y. C. Alteration of the pharmacokinetics of high-dose ara-C by its metabolite, high ara-U in patients with acute leukemia. J Clin Oncol. 1983 Dec;1(12):763–771. doi: 10.1200/JCO.1983.1.12.763. [DOI] [PubMed] [Google Scholar]
  7. Cass C. E., Gaudette L. A., Paterson A. R. Mediated transport of nucleosides in human erythrocytes. Specific binding of the inhibitor nitrobenzylthioinosine to nucleoside transport sites in the erythrocyte membrane. Biochim Biophys Acta. 1974 Apr 12;345(1):1–10. doi: 10.1016/0005-2736(74)90239-9. [DOI] [PubMed] [Google Scholar]
  8. Chabner B. A., Hande K. R., Drake J. C. Ara-C metabolism: implications for drug resistance and drug interactions. Bull Cancer. 1979;66(1):89–92. [PubMed] [Google Scholar]
  9. Cheng Y. C., Domin B., Lee L. S. Human deoxycytidine kinase. Purification and characterization of the cytoplasmic and mitochondrial isozymes derived from blast cells of acute myelocytic leukemia patients. Biochim Biophys Acta. 1977 Apr 12;481(2):481–492. doi: 10.1016/0005-2744(77)90281-9. [DOI] [PubMed] [Google Scholar]
  10. Cohen A., Ullman B., Martin D. W., Jr Characterization of a mutant mouse lymphoma cell with deficient transport of purine and pyrimidine nucleosides. J Biol Chem. 1979 Jan 10;254(1):112–116. [PubMed] [Google Scholar]
  11. Coleman C. N., Stoller R. G., Drake J. C., Chabner B. A. Deoxycytidine kinase: properties of the enzyme from human leukemic granulocytes. Blood. 1975 Nov;46(5):791–803. [PubMed] [Google Scholar]
  12. Drahovsky D., Kreis W. Studies on drug resistance. II. Kinase patterns in P815 neoplasms sensitive and resistant to 1-beta-D-arabinofuranosylcytosine. Biochem Pharmacol. 1970 Mar;19(3):940–944. doi: 10.1016/0006-2952(70)90259-5. [DOI] [PubMed] [Google Scholar]
  13. Duggleby R. G. A nonlinear regression program for small computers. Anal Biochem. 1981 Jan 1;110(1):9–18. doi: 10.1016/0003-2697(81)90104-4. [DOI] [PubMed] [Google Scholar]
  14. Furth J. J., Cohen S. S. Inhibition of mammalian DNA polymerase by the 5'-triphosphate of 1-beta-d-arabinofuranosylcytosine and the 5'-triphosphate of 9-beta-d-arabinofuranoxyladenine. Cancer Res. 1968 Oct;28(10):2061–2067. [PubMed] [Google Scholar]
  15. Gale R. P. Advances in the treatment of acute myelogenous leukemia. N Engl J Med. 1979 May 24;300(21):1189–1199. doi: 10.1056/NEJM197905243002105. [DOI] [PubMed] [Google Scholar]
  16. Harris A. L., Grahame-Smith D. G. The relationship of Ara-C metabolism in vitro to therapeutic response in acute myeloid leukaemia. Cancer Chemother Pharmacol. 1982;9(1):30–35. doi: 10.1007/BF00296758. [DOI] [PubMed] [Google Scholar]
  17. Heichal O., Ish-Shalom D., Koren R., Stein W. D. The kinetic dissection of transport from metabolic trapping during substrate uptake by intact cells. Uridine uptake by quiescent and serum-activated Nil 8 hamster cells and their murine sarcoma virus-transformed counterparts. Biochim Biophys Acta. 1979 Feb 20;551(1):169–186. doi: 10.1016/0005-2736(79)90363-8. [DOI] [PubMed] [Google Scholar]
  18. Ho D. H., Frei E., 3rd Clinical pharmacology of 1-beta-d-arabinofuranosyl cytosine. Clin Pharmacol Ther. 1971 Nov-Dec;12(6):944–954. doi: 10.1002/cpt1971126944. [DOI] [PubMed] [Google Scholar]
  19. Ives D. H., Durham J. P. Deoxycytidine kinase. 3. Kinetics and allosteric regulation of the calf thymus enzyme. J Biol Chem. 1970 May 10;245(9):2285–2294. [PubMed] [Google Scholar]
  20. Khym J. X. An analytical system for rapid separation of tissue nucleotides at low pressures on conventional anion exchangers. Clin Chem. 1975 Aug;21(9):1245–1252. [PubMed] [Google Scholar]
  21. Kufe D. W., Major P. P., Egan E. M., Beardsley G. P. Correlation of cytotoxicity with incorporation of ara-C into DNA. J Biol Chem. 1980 Oct 10;255(19):8997–8900. [PubMed] [Google Scholar]
  22. Kufe D., Spriggs D., Egan E. M., Munroe D. Relationships among Ara-CTP pools, formation of (Ara-C)DNA, and cytotoxicity of human leukemic cells. Blood. 1984 Jul;64(1):54–58. [PubMed] [Google Scholar]
  23. Lithotripsy. Health and Public Policy Committee, American College of Physicians. Ann Intern Med. 1985 Oct;103(4):626–629. [PubMed] [Google Scholar]
  24. Major P. P., Egan E. M., Beardsley G. P., Minden M. D., Kufe D. W. Lethality of human myeloblasts correlates with the incorporation of arabinofuranosylcytosine into DNA. Proc Natl Acad Sci U S A. 1981 May;78(5):3235–3239. doi: 10.1073/pnas.78.5.3235. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Momparler R. L., Fischer G. A. Mammalian deoxynucleoside kinase. I. Deoxycytidine kinase: purification, properties, and kinetic studies with cytosine arabinoside. J Biol Chem. 1968 Aug 25;243(16):4298–4304. [PubMed] [Google Scholar]
  26. Mulder J. H., Harrap K. R. Cytosine arabinoside uptake by tumour cells in vitro. Eur J Cancer. 1975 May;11(5):373–379. doi: 10.1016/0014-2964(75)90066-3. [DOI] [PubMed] [Google Scholar]
  27. Paterson A. R., Kolassa N., Cass C. E. Transport of nucleoside drugs in animal cells. Pharmacol Ther. 1981;12(3):515–536. doi: 10.1016/0163-7258(81)90096-6. [DOI] [PubMed] [Google Scholar]
  28. Paterson A. R., Yang S. E., Lau E. Y., Cass C. E. Low specificity of the nucleoside transport mechanism of RPMI 6410 cells. Mol Pharmacol. 1979 Nov;16(3):900–908. [PubMed] [Google Scholar]
  29. Plagemann P. G., Marz R., Wohlhueter R. M. Transport and metabolism of deoxycytidine and 1-beta-D-arabinofuranosylcytosine into cultured Novikoff rat hepatoma cells, relationship to phosphorylation, and regulation of triphosphate synthesis. Cancer Res. 1978 Apr;38(4):978–989. [PubMed] [Google Scholar]
  30. Rustum Y. M., Preisler H. D. Correlation between leukemic cell retention of 1-beta-D-arabinofuranosylcytosine 5'-triphosphate and response to therapy. Cancer Res. 1979 Jan;39(1):42–49. [PubMed] [Google Scholar]
  31. Schrecker A. W. Metabolism of 1-beta-D-arabinofuranosylcytosine in leukemia L1210: nucleoside and nucleotide kinases in cell-free extracts. Cancer Res. 1970 Mar;30(3):632–641. [PubMed] [Google Scholar]
  32. Tattersall M. H., Ganeshaguru K., Hoffbrand A. V. Mechanisms of resistance of human acute leukaemia cells to cytosine arabinoside. Br J Haematol. 1974 May;27(1):39–46. doi: 10.1111/j.1365-2141.1974.tb06772.x. [DOI] [PubMed] [Google Scholar]
  33. Weinstein H. J., Griffin T. W., Feeney J., Cohen H. J., Propper R. D., Sallan S. E. Pharmacokinetics of continuous intravenous and subcutaneous infusions of cytosine arabinoside. Blood. 1982 Jun;59(6):1351–1353. [PubMed] [Google Scholar]
  34. White J. C., Hines L. H., Rathmell J. P. Inhibition of 1-beta-D-arabinofuranosylcytosine transport and net accumulation by teniposide and etoposide in Ehrlich ascites cells and human leukemic blasts. Cancer Res. 1985 Jul;45(7):3070–3075. [PubMed] [Google Scholar]
  35. White J. C., Mikulecky D. C. Application of network thermodynamics to the computer modeling of the pharmacology of anticancer agents: a network model for methotrexate action as a comprehensive example. Pharmacol Ther. 1981;15(2):251–291. doi: 10.1016/0163-7258(81)90045-0. [DOI] [PubMed] [Google Scholar]
  36. Wiley J. S., Jones S. P., Sawyer W. H. Cytosine arabinoside transport by human leukaemic cells. Eur J Cancer Clin Oncol. 1983 Aug;19(8):1067–1074. doi: 10.1016/0277-5379(83)90029-9. [DOI] [PubMed] [Google Scholar]
  37. Wiley J. S., Jones S. P., Sawyer W. H., Paterson A. R. Cytosine arabinoside influx and nucleoside transport sites in acute leukemia. J Clin Invest. 1982 Feb;69(2):479–489. doi: 10.1172/JCI110472. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Wiley J. S., Taupin J., Jamieson G. P., Snook M., Sawyer W. H., Finch L. R. Cytosine arabinoside transport and metabolism in acute leukemias and T cell lymphoblastic lymphoma. J Clin Invest. 1985 Feb;75(2):632–642. doi: 10.1172/JCI111741. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Wohlhueter R. M., Marz R., Plagemann P. G. Thymidine transport in cultured mammalian cells. Kinetic analysis, temperature dependence and specificity of the transport system. Biochim Biophys Acta. 1979 May 17;553(2):262–283. doi: 10.1016/0005-2736(79)90231-1. [DOI] [PubMed] [Google Scholar]

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