title: Use of the comet assay to detect hypoxic cells in murine tumours and normal tissues exposed to bioreductive drugs.
abstract: The alkaline comet assay was applied to individual cells from mice exposed to two bioreductive drugs, tirapazamine and RSU 1069, with the goal of comparing DNA damage to tumours and normal tissues. More DNA single-strand breaks (SSBs) and a greater heterogeneity in DNA damage were observed in tumour cells than in spleen and marrow cells of mice exposed to 10-100 mg/kg tirapazamine, consistent with the presence of hypoxic cells and the greater bioreductive capacity of tumours. In mice injected with 25-200 mg/kg RSU 1069, aerobic cells exhibited large numbers of SSBs while toxic DNA interstrand crosslinks were produced only in hypoxic cells. Cells from bone marrow and spleen showed extensive numbers of SSBs, but minimal crosslinking compared to tumours where 10-20% of cells were heavily crosslinked. DNA damage produced by these two bioreductive drugs may be useful in estimating the range of individual cell oxygen contents within tumours and normal tissues.
authors.affiliation: Medical Biophysics Department, British Columbia Cancer Research Centre, Vancouver, Canada.
authors.forename: P L
authors.initials: PL
authors.lastname: Olive
mesh_terms: D000332:Aerobiosis; D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology*; D001853:Bone Marrow / Q000187:drug effects / Q000473:pathology / Q000528:radiation effects; D002294:Carcinoma, Squamous Cell / Q000473:pathology*; D015687:Cell Hypoxia; D004249:DNA Damage; D004277:DNA, Single-Stranded / Q000032:analysis / Q000187:drug effects / Q000528:radiation effects; D004307:Dose-Response Relationship, Radiation; D004587:Electrophoresis, Agar Gel / Q000379:methods; D006863:Hydrogen-Ion Concentration; D008297:Male; D051379:Mice; D008809:Mice, Inbred C3H; D008920:Misonidazole / Q000031:analogs & derivatives* / Q000494:pharmacology / Q000633:toxicity; D009928:Organ Specificity; D010084:Oxidation-Reduction; D011838:Radiation-Sensitizing Agents / Q000494:pharmacology*; D013154:Spleen / Q000187:drug effects / Q000473:pathology / Q000528:radiation effects; D000077704:Tirapazamine; D014227:Triazines / Q000494:pharmacology* / Q000633:toxicity
chemical_list: D000970:Antineoplastic Agents; D004277:DNA, Single-Stranded; D011838:Radiation-Sensitizing Agents; D014227:Triazines; D000077704:Tirapazamine; C041712:1-(2-nitro-1-imidazolyl)-3-aziridino-2-propanol; D008920:Misonidazole