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Measurement of deoxyuridine triphosphate and thymidine triphosphate in the extracts of thymidylate synthase-inhibited cells using a modified DNA polymerase assay.
Horowitz, R W; Zhang, H; Schwartz, E L; Ladner, R D; Wadler, S.
Afiliação
  • Horowitz RW; Department of Oncology, Montefiore Medical Center, and the Albert Einstein Cancer Center, Bronx, NY 10467, U.S.A.
Biochem Pharmacol ; 54(5): 635-8, 1997 Sep 01.
Article em En | MEDLINE | ID: mdl-9337081
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 dUTPdTTP 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 dUTPdTTP ratio at 2 hr after a 2-hr exposure to ZD1694 at concentrations of 0.05 to 0.2 microM 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.
Assuntos
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Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Quinazolinas / Tiofenos / Timidilato Sintase / Nucleotídeos de Timina / Nucleotídeos de Desoxiuracil / Antimetabólitos Antineoplásicos Limite: Humans Idioma: En Ano de publicação: 1997 Tipo de documento: Article
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Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Quinazolinas / Tiofenos / Timidilato Sintase / Nucleotídeos de Timina / Nucleotídeos de Desoxiuracil / Antimetabólitos Antineoplásicos Limite: Humans Idioma: En Ano de publicação: 1997 Tipo de documento: Article