Effect of Uridine on the Metabolism of 5-Fluorouracil in the CD8F1 Murine Mammary Carcinoma System

Abstract

The effect of uridine on the incorporation of 5-fluorouracil into RNA and the inhibition of DNA synthesis by the FdUMP block of thymidylate synthetase was studied in the CD8F1 murine mammary carcinoma system. The administration of exogenous uridine resulted in about a one third reduction of 5-fluorouracil in RNA of tumor and normal tissues. However, unlike thymidine, uridine was unable to reverse the early, partial inhibition of DNA synthesis. The amount of fluorouridine nucleotides and (5-fluorouracil)RNA formed in various tissues correlates with the level of orotate phosphoribosyl transferase activity suggesting that the major pathway for activation of 5-fluorouracil to nucleotide form in these tissues is via phosphoribosyl transferase. Enzyme preparations from three different murine tumors convert about 15 times as much 5-fluorouracil to FUMP as they do uracil to UMP. In contrast, the ratio of FUMP to UMP formed in enzyme preparations from gut and bone marrow is lower, 2–6 fold. However, in none of these tissues was the in vitro conversion of 5-fluorouracil to FUMP or incorporation into RNA substantially inhibited by uracil. Examination of tumor, gut and bone marrow uridine nucleotide pools showed that the thymidine-uridine-5-fluorouracil schedule does increase uridine nucleotide pools. Thus, the reduction in 5-fluorouracil in RNA is probably not due to inhibition of the conversion of 5-fluorouracil to FUMP by uracil (derived from phosphorylase cleavage of uridine) but, rather, is probably due to the elevated levels of UTP. We conclude that the protection from 5-fluorouracil toxicity afforded by the addition of uridine is due to the reduction in 5-fluorouracil in RNA rather than by reversal of the FdUMP block on thymidylate synthetase.