Comparison between the binding of [3H]misonidazole and AF-2 in Chinese hamster V79 spheroids.
Binding of two hypoxia probes, [3H]misonidazole and AF-2 (2-(2-Furyl)-3-(5-nitro-2-furyl)acrylamide), was compared in Chinese hamster V79 spheroids incubated under different oxygen concentrations. Fluorescence-activated cell sorting based on Hoechst 33342 penetration was used to obtain populations of cells from different depths within the spheroid, and sorted cells were analyzed by cytofluorometry for AF-2 content and by liquid scintillation counting for [3H]misonidazole content. The patterns of AF-2 and misonidazole binding were very similar, with about 20-fold more localization of both drugs in anoxic compared to aerobic cells. Similar results were obtained when cells were sorted on the basis of AF-2 rather than Hoechst 33342 fluorescence. When mean cellular fluorescence of AF-2 was plotted versus cpm misonidazole/cell for different oxygen tensions, it appeared that oxygen was equally effective in inhibiting AF-2 and misonidazole binding. Internal cells of anoxic spheroids bound about twice as much AF-2 and misonidazole as external cells, apparently due to an increased rate of nitroreduction by chronically hypoxic cells. AF-2 was found to enhance the retention of misonidazole in oxic and hypoxic spheroids when both drugs were present.
D000818:Animals; D002460:Cell Line; D002469:Cell Separation; D006224:Cricetinae; D003412:Cricetulus; D005434:Flow Cytometry; D005668:Furylfuramide / Q000378:metabolism; D008920:Misonidazole / Q000378:metabolism; D008954:Models, Biological; D009369:Neoplasms / Q000473:pathology*; D009581:Nitrofurans; D010100:Oxygen; D012588:Scintillation Counting
D009581:Nitrofurans; D005668:Furylfuramide; D008920:Misonidazole; D010100:Oxygen