Abstract
New inhibitors of the enzyme thymidylate synthase (TS) are now reaching clinical application. Alteration of the dUTP: dTTP ratio may be critical to TS inhibition-induced tumor cell death. The DNA polymerase assay with modification was used to rapidly and sensitively measure dUTP, dTTP, and dUTP:dTTP ratios in cell extracts of HT29 human colon carcinoma cells treated with the specific TS inhibitor ZD1694 [N-(5-[N-(3,4-dihydro-2-methyl-4 -oxoquinazolin-6-ylmethyl)-N-methylamino]-2-thenoyl)-L-glutamic acid]. These results revealed an increase in the dUTP:dTTP ratio at 2 hr after a 2-hr exposure to ZD1694 at concentrations of 0.05 to 0.2 μM with significant normalization at 16 hr after a 2-hr exposure despite evidence of continued TS inhibition. This assay is highly sensitive and reproducible for levels of dUTP and is less labor intensive than traditional assays.
Original language | English (US) |
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Pages (from-to) | 635-638 |
Number of pages | 4 |
Journal | Biochemical Pharmacology |
Volume | 54 |
Issue number | 5 |
DOIs | |
State | Published - Sep 1 1997 |
Keywords
- Antifolate
- Colon cancer
- DNA polymerase
- Deoxynucleotides
- Thymidylate synthase
ASJC Scopus subject areas
- Biochemistry
- Pharmacology