Cancer Chemotherapy and Pharmacology

, Volume 24, Issue 4, pp 251–255 | Cite as

A study of the mechanisms of cytotoxicity of Ara-C on three human leukemic cell lines

  • J. Zittoun
  • J. Marquet
  • J. C. David
  • D. Maniey
  • R. Zittoun
Original Articles Cytotoxicity of Ara-C, Human Leukemia Cell
Received:
Accepted:

Summary

The main biochemical determinants involved in cytosine arabinoside (Ara-C) metabolism were studied in one lymphoblastic (Reh) and two myeloid (HL60 and K562) human leukemic cell lines exhibiting various sensitivities to Ara-C, Reh being the most and HL60 the least sensitive. The level of intracellular Ara-C accumulation and Ara-CTP formation was far more important in Reh cells than in myeloid cell lines but was not closely related to deoxycytidine kinase activity or to deoxycytidine triphosphate pool size. The level of Ara-C incorporated into DNA was similar in the three cell lines. Ara-CTP formation correlated better with the cytotoxicity to clonogenic cells than did Ara-C incorporation into DNA. DNA polymerase α was moderately inhibited to various degrees, depending on the cell line; this moderate inhibition does not seem sufficient to explain the inhibition of DNA synthesis. The activity of DNA ligase, the enzyme joining the Okazaki fragments, which was not detected in Reh cells, was strongly inhibited by Ara-C in HL60 and to a lesser degree, in K562 cells. The inhibition of DNA ligase probably also contributes to the inhibition of DNA synthesis and, thus, to the cytotoxic effect of Ara-C and may explain the smaller size of DNA fragments observed following Ara-C treatment. The variations in each critical determinant observed in these three cell lines increase the complexity and plurality of the mechanisms of Ara-C action.

Keywords

K562 Cell Deoxycytidine Arabinoside Cytosine Arabinoside Clonogenic Cell 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.
This is a preview of subscription content, log in to check access.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Bhalla K, MacLaughlin W, Cole J, Arlin Z, Baker M, Graham G, Grant S (1987) Deoxycytidine preferentially protects normal versus leukemic myeloid progenitor cells from cytosine arabinoside-mediated cytotoxicity. Blood 70:561Google Scholar
  2. 2.
    Bremerskov V (1975) Inhibition of DNA synthesis by beta-d-arabinofuranosylcytosine. Biomedicine 23:217Google Scholar
  3. 3.
    Collins SJ, Gallo RC, Gallagher RE (1977) Continuous growth and differentiation of human myeloid leukemic cells in suspension culture. Nature 270:325Google Scholar
  4. 4.
    David JC, Zittoun R, Bassez T, Maniey D, Rusquet R, Bonhommet M, Le Prise PY, Thevenin D, Suberville AM, Marie JP (1985) DNA ligases in human leukemia. Leuk Res 9:851Google Scholar
  5. 5.
    DePamphilis ML, Wassarman PM (1980) Replication of eukaryotic chromosomes: a close-up of the replication fork. Ann Rev Biochem 49:627Google Scholar
  6. 6.
    Fox RM, Piddington SK, Tripp EH, Dudman NP, Tattersall MHN (1979) Thymidine sensitivity of cultured leukaemic lymphocytes. Lancet II:391Google Scholar
  7. 7.
    Fram RJ, Kufe DW (1982) DNA strand breaks caused by inhibitors of DNA synthesis: 1-β-d-arabinofuranosylcytosine and aphidicolin. Cancer Res 42:4050Google Scholar
  8. 8.
    Furth JJ, Cohen SS (1968) Inhibition of mammalian DNA polymerase by the 5′-triphosphate of 1-β-d-arabinofuranosylcytosine and the 5′-triphosphate of 9-β-d-arabinofuranosyladenine. Cancer Res 28:2061Google Scholar
  9. 9.
    Graham FL, Whitmore GF (1970) Studies in mouse L-cells on the incorporation of 1-β-d-arabinofuranosylcytosine into DNA and on inhibition of DNA polymerase by 1-β-d-arabinofuranosylcytosine 5′-triphosphate. Cancer Res 30:2618Google Scholar
  10. 10.
    Johnson RT, Collins ARS, Squires S, Mullinger AM, Elliot GC, Downes CS, Rasko I (1987) DNA repair under stress. J Cell Sci [Suppl] 6:263Google Scholar
  11. 11.
    Kufe DW, Major PP (1982) Studies on the mechanism of action of cytosine arabinoside. Med Pediatr Oncol 1 [Suppl]:26Google Scholar
  12. 12.
    Kufe DW, Major PP, Egan EE, Beardsley GP (1980) Correlation of cytotoxicity with incorporation of Ara-C into DNA. J Biol Chem 255:8997Google Scholar
  13. 13.
    Leclerc JM, Grise-Miron L, Labonte L (1987) Inhibition of DNA polymerase by Ara-CMP in the presence of a regulatory protein extracted from human promyelocytic leukemic cells (HL 60). Semin Oncol 14 (2) [Suppl 1]:226Google Scholar
  14. 14.
    Lehman R (1974) DNA ligase: structure, mechanism and function. Science 186:790Google Scholar
  15. 15.
    Lozzio CB, Lozzio BB (1975) Human chronic myelogenous leukemia cell-line with positive Philadelphia chromosome. Blood 45:321Google Scholar
  16. 16.
    Lindell TJ, Weinberg F, Morris PW, Roeder RG, Rutter WJ (1970) Specific inhibition of nuclear polymerase II by α amanitin. Science 170:425Google Scholar
  17. 17.
    Major PP, Egan EM, Herrick DJ, Kufe DW (1982) Effect of Ara-C incorporation on deoxyribonucleic acid synthesis in cells. Biochem Pharmacol 31:2937Google Scholar
  18. 18.
    Modrich P, Lehman IR (1970) Enzymatic joining of polynucleotides: IX. A simple and rapid assay of polynucleotide joining ligase by measurement of circle formation from linear deoxyadenylate-deoxythymidylate. J Biol Chem 245:3626Google Scholar
  19. 19.
    Momparler RL (1969) Effect of cytosine arabinoside 5′-triphosphate on mammalian DNA polymerase. Biochem Biophys Res Commun 34:245Google Scholar
  20. 20.
    Momparler RL (1972) Kinetic and template studies with 1-β-d-arabinofuranosylcytosine 5′-triphosphate and mammalian deoxyribonucleic acid polymerase. Mol Pharmacol 8:182Google Scholar
  21. 21.
    Piall EM, Aherne GW, Marks VW (1979) A radioimmunoassay for cytosine arabinoside. Br J Cancer 40:548Google Scholar
  22. 22.
    Robert De Saint Vincent B, Buttin G (1973) Studies on 1-β-d-arabinofuranosylcytosine-resistant mutants of Chinese hamster fibroblasts: a mitochondrial deoxycytidine kinase devoid of activity on arabinocytosine. Eur J Biochem 37:481Google Scholar
  23. 23.
    Robichaud NJ, Fram RJ (1987) Potentiation of Ara-C induced cytotoxicity by hydroxyurea in LoVo colon carcinoma cells. Biochem Pharmacol 18:1673Google Scholar
  24. 24.
    Rosenfeld C, Goutner A, Choquet C, Venuat AM, Keyibanda B, Pico JL, Greaves MF (1977) Phenotypic characterisation of a unique non-T, non-B acute lymphoblastic leukemia cell line. Nature 267:841Google Scholar
  25. 25.
    Soderhall S (1976) DNA ligase during rat liver regeneration. Nature 260:640Google Scholar
  26. 26.
    Skoog L (1970) An enzymatic method for the determination of dCTP and dGTP in picomole amounts. Eur J Biochem 17:202Google Scholar
  27. 27.
    Tsang Lee MYW, Byrnes JJ, Downey KM, So AG (1980) Mechanism of inhibition of deoxyribonucleic acid synthesis by 1-β-d-arabinofuranosyladenosine triphosphate and its potentiation by 6-mercaptopurine ribonucleoside 5′-triphosphate. Biochemistry 19:210Google Scholar
  28. 28.
    Waqar MA, Burgoyne LA, Atkinson MR (1971) Deoxyribonucleic acid synthesis in mammalian nuclei. Incorporation of deoxyribonucleotides and chain-terminated nucleotide analogues. Biochem J 121:803Google Scholar
  29. 29.
    Wiley JS, Jones SP, Sawyer WH, Patterson ARP (1982) Cytosine arabinoside influx and nucleoside transport sites in acute leukemia. J Clin Invest 69:259Google Scholar
  30. 30.
    Wist E, Krokan H, Prydz H (1976) Effect of 1-β-d-arabinofuranosylcytosine triphosphate on DNA synthesis in isolated HeLa cell nuclei. Biochemistry 15:1825Google Scholar

Copyright information

© Springer-Verlag 1989

Authors and Affiliations

  • J. Zittoun
    • 1
  • J. Marquet
    • 1
  • J. C. David
    • 2
  • D. Maniey
    • 2
  • R. Zittoun
    • 3
  1. 1.Laboratoire Central d'HématologieHôpital Henri MondorCréteilFrance
  2. 2.Laboratoire de Biochimie du DéveloppementUniversité de RennesFrance
  3. 3.Service d'HématologieParisFrance

Personalised recommendations