Summary
Changes are reported in [14C]-formate incorporation into nucleic acids and protein of Ehrlich ascites tumor cells during exposure to methotrexate (MTX) and fluoropyrimidines. The rate of [14C]-formate incorporation into RNA, DNA, and protein in the presence of only MTX was inhibited by 82%, 91%, and 75% respectively, when compared with control rates. However, in the presence of 5-fluorodeoxyuridine (FdUrd) plus MTX, formate incorporation into RNA, DNA, and protein was inhibited by 67%, 85%, and 66%. Incubation of cells in vitro with [3H]-dihydrofolate (DHF) results in its rapid conversion to [3H]-tetrahydrofolate (THF). The THF/DHF ratio from the soluble fraction of cells that were incubated with [3H]-DHF was 43% greater in the presence of FdUrd and MTX than in the presence of MTX alone. As the rate of [3H]-dUrd incorporation into DNA was reduced by 88% and 99% by pretreating cells with 0.1 μM and 1 μM FdUrd, respectively, the inhibitory effect of MTX on [14C]-formate incorporation into (a) RNA was decreased by 63% and 46%; (b) DNA was decreased by 74% and 61%; and (c) protein was decreased by 63% and 32%. These data suggest that fluoropyrimidines can antagonize the effects of MTX on purines or nucleic acid synthesis and protein synthesis by preventing the consumption of THF for dTMP synthesis.
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Abbreviations
- MTX:
-
methotrexate
- FdUrd:
-
5-fluorodeoxyuridine
- Fura:
-
5-fluorouracil
- FdUMP:
-
5-fluorodeoxyuridine monophosphate
- dUrd:
-
deoxyuridine
- dUMP:
-
deoxyuridine monophosphate
- dTMP:
-
thymidylate
- DHF:
-
dihydrofolate
- THF:
-
tetrahydrofolate
- 5,10-CH2THF:
-
5,10-methylene tetrahydrofolate
- DHFR:
-
dihydrofolate reductase
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Bowen, D., Fölsch, E. & Guernsey, L.A. The interaction between fluoropyrimidines and methotrexate, and [14C]-formate incorporation into nucleic acids and protein. Cancer Chemother. Pharmacol. 4, 111–116 (1980). https://doi.org/10.1007/BF00254031
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DOI: https://doi.org/10.1007/BF00254031