Semiautomated 14C-thymidine incorporation assay for in vitro screening of anticancer drug.
We developed a rapid screening test for anticancer drugs by using 14C-thymidine (14C-TdR), a microculture filtration plate and a radiochromatoscanner. Mitomycin C (MMC), tamoxifen and 5-fluorouracil (5FU) were used as the anticancer drugs against four human gastric cancer cell lines. The rates of cell inactivation (14C-TdR uptake) determined with the radiochromatoscanner were almost the same as those determined with the liquid scintillation counter. Comparing the rate of cell inactivation obtained by our method with that obtained by proliferative activity of cells derived from the cell count, both assays showed the approximately same results by tamoxifen and MMC but the rates of cell inactivation by 5FU in two cell lines obtained by the 14C-TdR uptake assay was considerably lower than those obtained by the cell count. These results show that the radio-labeled DNA precursor uptake assay is not suitable for metabolic inhibitors of DNA synthesis. Ninety six samples on a plate, however, were assayed semiautomatically and rapidly just as well as in the tetrazolium-based calorimetric (MTT) assay. Therefore, our 14C-TdR uptake assay system is useful for the cancer chemotherapeutic agents except the metabolic inhibitors of DNA precursors.
Miyazaki
N
N
Department of Radioisotope, Tokyo Institute of Psychiatry.
Ohkura
H
H
Kajimura
N
N
Okazaki
N
N
D000230:Adenocarcinoma / Q000473:pathology; D000970:Antineoplastic Agents / Q000494:pharmacology*; D002250:Carbon Radioisotopes; D018279:Carcinoma, Signet Ring Cell / Q000473:pathology; D002455:Cell Division; D004354:Drug Screening Assays, Antitumor / Q000379:methods*; D005472:Fluorouracil / Q000494:pharmacology; D006801:Humans; D016685:Mitomycin / Q000494:pharmacology; D013274:Stomach Neoplasms / Q000473:pathology*; D013629:Tamoxifen / Q000494:pharmacology; D013936:Thymidine; D014407:Tumor Cells, Cultured
D000970:Antineoplastic Agents; D002250:Carbon Radioisotopes; D013629:Tamoxifen; D016685:Mitomycin; D005472:Fluorouracil; D013936:Thymidine