Methotrexate Polyglutamates in Cultured Human Cells

  • David S. Rosenblatt
  • V. Michael Whitehead
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 163)

Abstract

Cultured human fibroblasts accumulated methotrexate polyglutamates to levels far in excess of the dihydrofolate reductase binding capacity. After four days in methotrexate-free medium the intracellular drug level dropped by 70% but nearly 80% of the remaining methotrexate was in the form of polyglutamates. Reduced folates prevented the accumulation of polyglutamates and the effects of methotrexate on deoxyuridine incorporation into DNA and cell growth if present along with methotrexate from the beginning of the incubation. However, the reduced folates were less effective if added to cells after a long exposure to methotrexate alone. Thymidine, glycine, and adenosine (GAT) prevent methotrexate toxicity only if maintained in the incubation medium. However, preincubation with methotrexate and GAT permits continued synthesis and accumulation of polyglutamates so that when the GAT and methotrexate were removed, toxicity from the retained methotrexate polyglutamates could be expressed. (2,4-diamino-5-(3′4′-dichlorophenyl)-6 methyl pyrimidine (DDMP), an antifol that does not form polyglutamate derivatives, inhibited deoxyuridine incorporation into DNA as long as the DDMP remained in the culture medium. Compared to what was seen with longer exposures to methotrexate, removal of DDMP resulted in a greater reversal of inhibition of deoxyuridine incorporation.

Keywords

Folinic Acid Dihydrofolate Reductase Exchangeable Fraction Confluent Fibroblast Great Reversal 
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.

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Copyright information

© Springer Science+Business Media New York 1983

Authors and Affiliations

  • David S. Rosenblatt
    • 1
  • V. Michael Whitehead
    • 1
  1. 1.MRC Genetics Group and The Penny Cole Hematology Research LaboratoryMcGill University-Montreal Children’s Hospital Research InstituteMontrealCanada

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