Use of Flow Microfluorometry in Detailed Analysis of Effects of Chemical Agents on Cell Cycle Progression

摘要: A technique has been developed for determining effects of chemotherapeutic agents on cell cycle progression in synchronized line CHO Chinese hamster cells grown vitro . The proctocol combines autoradiographic and cell-enumeration techniques with analysis DNA distribution patterns by means the Los Alamos flow microfluorometer. were accumulated a state G1 arrest growth isoleucine-deficient medium; then resuspended fresh, complete medium containing isoleucine plus test agent to reinitiate traverse presence drug. Ten hr later, microfluorometric population contents cultures yielded fractions G1, S, G2 or M. For reversible effects, confirmatory data obtained after resuspension drug-free thymidine-3H; aliquots removed at intervals determination capacity, even slowly nonprogressing populations. Four differing examined, namely, hydroxyurea, cytosine arabinoside, bleomycin, camptothecin. Both hydroxyurea (10-3 m) arabinoside (5 µg/ml) grossly decreased rate into S phase, resulting an accumulation G1-S boundary. Neither completely prevented from initiating synthesis. Bleomycin (100 allowed initiation completion genome replication occur nearly normal rate, but most lost capacity enter mitosis. Camptothecin (1 reduced overall few replicate complement DNA. relationship these findings chemotherapy regimen design is discussed.