Studies of Formation and Efflux of Methotrexate Polyglutamates with Cultured Hepatic Cells

  • John Galivan
  • Malgorzata Balinska
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 163)


Methotrexate polyglutamates are extensively synthesized when cultured hepatocytes and H35 hepatoma cells are exposed to micro-molar concentrations of methotrexate. The predominant species found within the cell have from two to four additional γ-linked glutamate residues. When either cell type containing a mixture of methotrexate and its polyglutamate derivatives is exposed to medium lacking methotrexate, there is a rapid release of methotrexate. This release has a (math) of 2 to 4 min and is apparently complete within 30 to 60 min. Methotrexate polyglutamates leave the cells much more slowly and appear to do so by two mechanisms. Although cleavage to methotrexate and subsequent efflux appears to be quantitatively the more important pathway, there is also a slow, finite loss of intact methotrexate polyglutamates from cells which exclude trypan blue. The T1/2 for the loss of methotrexate polyglutamates by both cell types, when placed in medium lacking methotrexate, is approximately 6 to 8 hr. These results, together with those of an earlier study (Galivan, J. (1980) Mol. Pharmacol. 17: 105–110), suggest that the polyglutamate derivatives are forms of methotrexate which are as cytotoxic as methotrexate but which offer a potentially greater capacity for cellular destruction because they are retained longer in the tissue.


Dihydrofolate Reductase Tetrahydrofolic Acid Methotrexate Polyglutamates Efficient Cell Kill Polyglutamate Derivative 
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.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Baugh CM, Krumdieck CL and Nair MG (1973) Biochem. Biophys. Res. Commun. 52:27–34.PubMedCrossRefGoogle Scholar
  2. 2.
    Covey JM (1980) Life Soi. 26:665–678.CrossRefGoogle Scholar
  3. 3.
    Jacobs SA, Adamson RH, Chabner BA, Derr CJ and Johns DG (1975) Biochen. Biophys. Res. Commun. 63:692–698.CrossRefGoogle Scholar
  4. 4.
    Whitehead VM (1977) Cancer Res. 37:408–412.PubMedGoogle Scholar
  5. 5.
    Galivan J (1980) Mol. Pharmacol. 17:105–110.PubMedGoogle Scholar
  6. 6.
    Schilsky RL, Bailey BD and Chabner BA (1980) Proc. Natl. Acad. Sci. 77:2919–2922.PubMedCrossRefGoogle Scholar
  7. 7.
    Goldman ID (1977) Cancer Treat. Rep. 61:549–558.PubMedGoogle Scholar
  8. 8.
    Sirotnak FM (1980) Pharmac. Ther. 8:71 – 103.CrossRefGoogle Scholar
  9. 9.
    Galivan J (1979) In Chemistry and Biology of Pteridines, Kisliuk RL and Brown GM (eds) Vol. 4, pp. 543–548, Elsevier, New York.Google Scholar
  10. 10.
    Galivan J (1979) Res. Commun. Chem. Pathol. Pharmcol. 24:571–582.Google Scholar
  11. 11.
    Galivan J, Tarentino A and Samsonoff W (1980) Ann. N.Y. Acad. Sci. 349:332–345.PubMedCrossRefGoogle Scholar
  12. 12.
    Tarentino AL and Galivan J (1980) .In Vitro 16:833–846.Google Scholar
  13. 13.
    Galivan J (1979) Cancer Res. 39:735–743.PubMedGoogle Scholar
  14. 14.
    Galivan J (1980) Arch. Biochem. Biophys. 206:113–121.CrossRefGoogle Scholar
  15. 15.
    Galivan J (1981) Cancer Res. 41: 1757–1762.PubMedGoogle Scholar
  16. 16.
    Balinska M, Galivan J and Coward JK (1981) Cancer Res. 41:2751–2756.PubMedGoogle Scholar
  17. 17.
    Galivan J (1981) Exp. Cell Res. 131:379–385.PubMedCrossRefGoogle Scholar
  18. 18.
    Laishes BA and Williams GM (1976) In Vitro 12: 821–832.Google Scholar
  19. 19.
    Miller MR, Casteilot JJ, Jr. and Pardee AB (1978) Biochemistry 17:1073–1078.PubMedCrossRefGoogle Scholar
  20. 20.
    Poser RG, Sirotnak FM and Chello PL (1980) Biochem. Pharmacol. 22:2701–2704.CrossRefGoogle Scholar
  21. 21.
    Yalowich JC, Fry DW and Goldman ID (1981) Proceed. Amer. Assoc. Cancer Res. 22:207.Google Scholar
  22. 22.
    Jolivet J, Schilsky RL and Chabner BA (1981) Proceed. Amer. Assoc. Cancer Res. 22:229.Google Scholar

Copyright information

© Springer Science+Business Media New York 1983

Authors and Affiliations

  • John Galivan
    • 1
  • Malgorzata Balinska
    • 1
  1. 1.Division of Laboratories and ResearchNew York State Department of HealthAlbanyUSA

Personalised recommendations