title: Photogenotoxicity of skin phototumorigenic fluoroquinolone antibiotics detected using the comet assay.
abstract: The fluoroquinolone (FQ) antibiotics photosensitize human skin to solar UV radiation and are reported to photosensitize tumor formation in mouse skin. As tumor initiation will not occur without genotoxic insult, we examined the potential of ciprofloxacin, lomefloxacin, fleroxacin, BAYy3118 (a recently developed monofluorinated quinolone) and a nalidixic acid to photosensitize DNA damage in V79 hamster fibroblasts in vitro. Cells were exposed to 37.5 kJ/m2 UVA (320-400 nm; glass filtered Sylvania psoralen + UVA (PUVA) tubes; calibrated Waldmann radiometer) at 4 degrees C in the presence of FQ and immediately afterwards embedded in agarose, lysed and placed in an electrophoretic field at pH 12. Under these denaturing conditions, the presence of DNA single-strand breaks (SSB), alkali-labile sites (ALS) and double-strand breaks (DSB) can be visualized as DNA migrating away from the nucleus (characteristic "comet" appearance) after staining with a specific fluorochrome. At FQ concentrations that induced minimal loss of cell viability (neutral red uptake assay) the compounds tested induced comets with a rank order of BAYy3118 > norfloxacin > ciprofloxacin > lomefloxacin > fleroxacin > nalidixic acid. If cells were incubated after treatment for 1 h at 37 degrees C, the comet score decreased, suggesting efficient removal of SSB/ALS/DSB. Addition of the DNA polymerase(alpha) inhibitor, aphidicolin, to cells treated with either ciprofloxacin alone or ciprofloxacin + UVA resulted in an accumulation of SSB due to the endo/exonuclease steps of excision repair. We have demonstrated that the FQ are photogenotoxic in mammalian cells but the FQ-photosensitized SSB are efficiently repaired. Preliminary evidence that ciprofloxacin photosensitizes the formation of DNA lesions warranting excision repair may indicate production of more mutagenic lesions.
authors.affiliation: Photobiology Unit, Ninewells Hospital, Dundee, Scotland.
authors.forename: H J
authors.initials: HJ
authors.lastname: Reavy
authors.forename: N J
authors.initials: NJ
authors.lastname: Traynor
authors.forename: N K
authors.initials: NK
authors.lastname: Gibbs
mesh_terms: D000818:Animals; D000890:Anti-Infective Agents / Q000633:toxicity*; D016590:Aphidicolin / Q000494:pharmacology; D006224:Cricetinae; D003412:Cricetulus; D004247:DNA / Q000528:radiation effects; D004260:DNA Repair; D004587:Electrophoresis, Agar Gel / Q000379:methods*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D024841:Fluoroquinolones; D006801:Humans; D051379:Mice; D009153:Mutagens / Q000633:toxicity*; D019384:Nucleic Acid Synthesis Inhibitors; D017319:Photosensitizing Agents / Q000633:toxicity*; D012867:Skin / Q000528:radiation effects*
chemical_list: D000890:Anti-Infective Agents; D004791:Enzyme Inhibitors; D024841:Fluoroquinolones; D009153:Mutagens; D019384:Nucleic Acid Synthesis Inhibitors; D017319:Photosensitizing Agents; D016590:Aphidicolin; D004247:DNA