title: Detection of oxaliplatin-induced DNA crosslinks in vitro and in cancer patients using the alkaline comet assay.
abstract: Oxaliplatin is frequently used in the therapy of cancer. In DNA, oxaliplatin induces, like cisplatin, the formation of crosslinks, which are commonly accepted as being responsible for the cytotoxicity of platinum agents. The detection of oxaliplatin-induced DNA crosslink formation and repair could be a good measure of assessing how a patient is responding to the agent. In this study, we used a validated modification of the alkaline comet assay for detecting the presence of these crosslinks in vitro and in cancer patients. The H460 tumour cell line was treated in vitro with a range of oxaliplatin and cisplatin doses, and the subsequent crosslink formation and repair compared between the two agents. In addition, lymphocytes from cancer patients undergoing oxaliplatin-based chemotherapy were assayed for the formation and repair of oxaliplatin-induced crosslinks. A dose-response was observed in the in vitro samples, with cisplatin producing more crosslinks than oxaliplatin at equimolar concentrations and lesions induced by both agents showing different repair efficiencies. Furthermore, evidence of crosslink formation and repair was observed in the peripheral blood lymphocytes of all cancer patients studied, along with the detection of interindividual variability in crosslink formation and repair efficiencies. To the best of our knowledge, this is the first time that oxaliplatin DNA crosslinks have been detected either in vitro or in patient samples using the alkaline comet assay. Due to its sensitivity, rapidity, small cell sample and low cost, the alkaline comet assay is a good method for the detection of oxaliplatin-induced crosslinks and their subsequent repair and, in future clinical studies, could prove to be a valuable tool in assessing/predicting a patient's response to chemotherapy.
authors.affiliation: Department of Cancer Studies and Molecular Medicine, University of Leicester, University Road, Leicester LE1 7RH, UK.
authors.forename: Gabriela M
authors.initials: GM
authors.lastname: Almeida
authors.forename: Tiago L
authors.initials: TL
authors.lastname: Duarte
authors.forename: William P
authors.initials: WP
authors.lastname: Steward
authors.forename: George D D
authors.initials: GD
authors.lastname: Jones
mesh_terms: D000368:Aged; D018287:Carcinoma, Large Cell / Q000188:drug therapy*; D045744:Cell Line, Tumor; D002945:Cisplatin / Q000494:pharmacology; D015179:Colorectal Neoplasms / Q000188:drug therapy*; D020552:Comet Assay / Q000379:methods; D003432:Cross-Linking Reagents / Q000494:pharmacology*; D004247:DNA / Q000235:genetics* / Q000378:metabolism; D004249:DNA Damage; D004260:DNA Repair; D004259:DNA-Directed DNA Polymerase / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D005260:Female; D006801:Humans; D008175:Lung Neoplasms / Q000188:drug therapy*; D008214:Lymphocytes / Q000166:cytology / Q000378:metabolism; D008297:Male; D008875:Middle Aged; D008956:Models, Chemical; D015233:Models, Statistical; D009362:Neoplasm Metastasis; D009369:Neoplasms / Q000235:genetics*; D009944:Organoplatinum Compounds / Q000494:pharmacology*; D000077150:Oxaliplatin; D012680:Sensitivity and Specificity; D013997:Time Factors; D000094769:DNA Polymerase iota
chemical_list: D003432:Cross-Linking Reagents; D009944:Organoplatinum Compounds; D000077150:Oxaliplatin; D004247:DNA; D004259:DNA-Directed DNA Polymerase; D002945:Cisplatin; D000094769:DNA Polymerase iota; C000722070:POLI protein, human