A non-radioactive sensitive assay to measure 5-fluorouracil incorporation into DNA of solid tumors.
نویسندگان
چکیده
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.
منابع مشابه
Quantification of 5-fluorouracil incorporation into RNA of human and murine tumors as measured with a sensitive gas chromatography-mass spectrometry assay.
5-Fluorouracil (5FU) can exert its cytotoxic activity by either inhibition of thymidylate synthase or incorporation into RNA. The extent and importance of the latter in tumors of patients are not clear, due to the lack of sensitive and reproducible methods. RNA from 5FU-treated human WiDr colon tumor cells was isolated and [(14)CL]5FU incorporation into RNA was measured by traditional scintilla...
متن کاملEffects of 5-fluorouracil on 5-fluorodeoxyuridine 5'-monophosphate and 2-deoxyuridine 5'-monophosphate pools, and DNA synthesis in solid mouse L1210 and rat Walker 256 tumors.
We have examined some of the factors determining the action of 5-fluorouracil (FUra) against a responsive and nonresponsive rodent tumor. In mice bearing the FUraresponsive solid L1210 tumor, [3H]deoxyuridine (dUrd) incorporation into tumor DNA ceased promptly after FUra (100 mg/kg i.p.), and recovery was delayed for more than 96 hr. Incorporation of [3H]dUrd into bone marrow and small intestin...
متن کاملMeasurement of DNA synthesis in leucocyte microcultures.
A microculture technique is described in which human peripheral blood leucocytes undergo DNA synthesis in response to specific and non-specific blastogenic factors. A sensitive Millipore filter assay technique is shown to be suitable for determining the incorporation of radioactive thymidine into the DNA of the stimulated cells in these microcultures. With these methods it is possible to measur...
متن کاملEffects of 5-Fluorouracil on 5-Fluorodeoxyuridine 5'-Monophosphate and 2-Deoxyuridine 5'-Monophosphate Pools, and DMA Synthesis in Solid Mouse L1210 and Rat Walker 256 Tumors1
We have examined some of the factors determining the action of 5-fluorouracil (FUra) against a responsive and nonresponsive rodent tumor. In mice bearing the FUraresponsive solid L1210 tumor, [3H]deoxyuridine (dUrd) incorporation into tumor DNA ceased promptly after FUra (100 mg/kg i.p.), and recovery was delayed for more than 96 hr. Incorporation of [3H]dUrd into bone marrow and small intestin...
متن کاملThe effect of 5-fluorouracil on DNA-thymine synthesis in human tumors.
eluent. Uptake patterns were highly individual, and large differences were noted both in the baseline uptakes and in the effect of 5-fluorouracil on precursor incorporation. Neither tissue of origin nor control uptake influenced the in vitro response to the drug. A decrease in formate incorporation was usually associated with a reciprocal increase in thymidine incorporation. Clinical data conce...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
عنوان ژورنال:
- Nucleosides, nucleotides & nucleic acids
دوره 23 8-9 شماره
صفحات -
تاریخ انتشار 2004