Role of Methotrexate Polyglutamates in Methotrexate- and Sequential Methotrexate-5-Fluorouracil-mediated Cell Kill

John J. McGuire, Enrico Mini, Pearl Hsieh, Joseph R. Bertino

Research output: Contribution to journalArticlepeer-review

34 Scopus citations

Abstract

The synthesis of methotrexate (MTX) polyglutamates has been investigated in a human leukemia cell line, CCRF-CEM. An increase in either the extracellular MTX concentration (0.1-10 μM) or exposure time (4-24 h) allowed a greater accumulation of polyglutamate derivatives. The species which predominated (di-, tri-, or tetraglutamate) was also time and concentration dependent. Derivatives up to the pentaglutamate were readily detectable, with hexaglutamate present under some conditions. Polyglutamate derivatives were preferentially retained when the cells were transferred to drug-free medium. MTX itself rapidly exited the cells under these conditions. In the absence of extracellular MTX, the preformed intracellular MTX polyglutamates continued to be elongated. Accumulation of MTX polyglutamates, particularly triglutamate and longer species, positively correlated with cytotoxicity to CCRF-CEM cells as measured by a clonogenic assay. Investigation of the methotrexate-fluorouracil interaction revealed no effect of 5-fluorouracil on MTX polyglutamate accumulation under various conditions where widely different degrees of cytotoxicity were found. The synergy observed when MTX precedes fluorouracil treatment is therefore not an effect on MTX polyglu-tamylation. MTX polyglutamates may, however, play an indirect role in this synergy.

Original languageEnglish (US)
Pages (from-to)6395-6400
Number of pages6
JournalCancer Research
Volume45
StatePublished - Dec 1 1985

All Science Journal Classification (ASJC) codes

  • Oncology
  • Cancer Research

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