The “anti-pyrimidine effect” of hypoxia and brequinar sodium (NSC 368390) is of consequence for tumor cell growth

Abstract

The rationale of the present study was to investigate the simultaneous effect of hypoxia and drugs with an “anti-pyrinudine effect” on tumor cell proliferation to evaluate putative changes in the sensitivity of cells to these kinds of chemotherapeutic treatment on reduced O 2 tension. Pyrimidine de novo biosynthesis, at the stage of respiratory chain-dependent dihydroorotate dehydrogenase, was found to be a biochemical target site for oxygen deficiency as well as for Brequinar Sodium TM (6-fluoro-2-(2'-nuoro-1,1'-biphenyl-4-yl)-3-methyl-4 quinoline carboxylic acid sodium salt) (Brequinar). Increasing drug concentrations (0.1–50 μM) reduced the proliferation rate of in vitro cultured Ehrlich ascites tumor cells ( ic 50 = 0.25 μM). Decreasing concentrations of O 2 reduced the proliferation rate (50% at ~3.5% O 2). Brequinar at 2.5 and 12.5 μM stimulated the incorporation of exogenous [ 14C]uridine into RNA to 140 and 190% of controls, respectively, as a result of active salvage pathways, whereas it decreased the incorporation of [ 14C]NaHCO 3 by the de novo pathway (to 20 and 5% of controls, respectively). Cells routinely grown in glucose-free, undine-supplemented medium were resistant to 12.5 μM of the drug. The complete growth pattern of the tumor cells (increase in cell number and protein, RNA and DNA content of cultures during a 24-hr culture period) was examined (i) on reducing the O 2 tension of the atmosphere stepwise from 20 to 1% O 2 (ii) on addition of 0.125 μM Brequinar; and (iii) under both conditions. The combination was found to give an additive inhibitory effect under moderate hypoxia (5–20% 0;) and a greater than additive effect if the oxygen tension was further reduced (1–5%).