title: A non-radioactive sensitive assay to measure 5-fluorouracil incorporation into DNA of solid tumors.
abstract: A non-radioactive method to determine 5-Fluorouracil (5FU) incorporation into DNA has been developed. Isolated DNA was enzymatically degraded to bases and the resulting 5FU was measured with standard gas-chromatography coupled to mass spectrometry (GC-MS) and compared with that of radioactive 5FU in a cell line. Incorporation into DNA of the murine Colon 26-B tumor treated with maximal tolerated doses of 5FU and fluorodeoxyuridine (FUdR) was maximal after 2 hour and was 15.4 and 71.0 fmol/microg DNA, respectively. After a plateau for about 3 days a decrease was observed to +/- 2 fmol/microg DNA after 10 days. The assay is very sensitive and reproducible and can be used in a clinical setting.
authors.affiliation: Department of Medical Oncology, VU University Medical Center, Amsterdam, The Netherlands.
authors.forename: P
authors.initials: P
authors.lastname: Noordhuis
authors.forename: U
authors.initials: U
authors.lastname: Holwerda
authors.forename: J A M
authors.initials: JA
authors.lastname: Van Laar
authors.forename: C L
authors.initials: CL
authors.lastname: Van der Wilt
authors.forename: G J
authors.initials: GJ
authors.lastname: Peters
mesh_terms: D000818:Animals; D000964:Antimetabolites, Antineoplastic / Q000493:pharmacokinetics*; D045744:Cell Line, Tumor; D003110:Colonic Neoplasms / Q000188:drug therapy; D004247:DNA / Q000187:drug effects* / Q000378:metabolism; D004354:Drug Screening Assays, Antitumor / Q000379:methods*; D005467:Floxuridine / Q000494:pharmacology*; D005472:Fluorouracil / Q000493:pharmacokinetics*; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods*; D051379:Mice; D009369:Neoplasms / Q000188:drug therapy*; D012680:Sensitivity and Specificity; D013997:Time Factors
chemical_list: D000964:Antimetabolites, Antineoplastic; D005467:Floxuridine; D004247:DNA; D005472:Fluorouracil