Detection of oxaliplatin-induced DNA crosslinks in vitro and in cancer patients using the alkaline comet assay
Detection of oxaliplatin-induced DNA crosslinks in vitro and in cancer patients using the alkaline comet assay
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DOI：
10.1016/j.dnarep.2005.09.010
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发表时间：
2006-02-03
期刊：
影响因子：
3.8
通讯作者：
Jones, GDD
中科院分区：
文献类型：
--
作者：
Almeida, GM;Duarte, TL;Jones, GDD
奥沙利铂常用于癌症的治疗。在DNA中，奥沙利铂和顺铂一样，可以诱导交联体的形成，这通常被认为是导致铂类药物细胞毒性的原因。检测奥沙利铂诱导的DNA交联形成和修复可能是评估患者对该药物的反应情况的一个很好的衡量标准。在这项研究中，我们使用了一种经过验证的改良碱性彗星试验来检测这些交联物在体外和癌症患者中的存在。在体外用不同剂量的奥沙利铂和顺铂处理H460肿瘤细胞系，并比较两种药物之间随后的交联物的形成和修复。此外，对接受以奥沙利铂为基础的化疗的癌症患者的淋巴细胞进行了检测，以确定奥沙利铂诱导的交叉连接的形成和修复。在体外标本中观察到了剂量效应，在等摩尔浓度下顺铂比奥沙利铂产生更多的交联物，两种药物诱导的损伤修复效率不同。此外，在所有被研究的癌症患者的外周血淋巴细胞中观察到了交联物形成和修复的证据，以及交联物形成和修复效率的个体间变异性的检测。据我们所知，这是第一次在体外或使用碱性彗星试验在患者样本中检测到奥沙利铂DNA交联物。由于其灵敏、快速、小细胞样本和低成本，碱性彗星试验是一种检测奥沙利铂诱导的交联链及其后续修复的良好方法，在未来的临床研究中，可能被证明是评估/预测患者对化疗的反应的有价值的工具。(C)2005 Elsevier B.V.保留所有权利。
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. (c) 2005 Elsevier B.V. All rights reserved.