Espina Marta, Corte-Rodríguez Mario, Aguado Leticia, Montes-Bayón María, Sierra Marta I, Martínez-Camblor Pablo, Blanco-González Elisa, Sierra L María
Dpt. of Functional Biology (Genetic Area) and Oncology University Institute (IUOPA), University of Oviedo, Oviedo 33006, Spain.
Metallomics. 2017 May 24;9(5):564-574. doi: 10.1039/c7mt00014f.
Cisplatin, one of the most extensively used metallodrugs in cancer treatment, presents the important drawback of patient resistance. This resistance is the consequence of different processes including those preventing the formation of DNA adducts and/or their quick removal. Thus, a tool for the accurate detection and quantitation of cisplatin-induced adducts might be valuable for predicting patient resistance. To prove the validity of such an assumption, highly sensitive plasma mass spectrometry (ICP-MS) strategies were applied to determine DNA adduct levels and intracellular Pt concentrations. These two metal-relative parameters were combined with an evaluation of biological responses in terms of genomic stability (with the Comet assay) and cell cycle progression (by flow cytometry) in four human cell lines of different origins and cisplatin sensitivities (A549, GM04312, A2780 and A2780cis), treated with low cisplatin doses (5, 10 and 20 μM for 3 hours). Cell viability and apoptosis were determined as resistance indicators. Univariate linear regression analyses indicated that quantitation of cisplatin-induced G-G intra-strand adducts, measured 1 h after treatment, was the best predictor for viability and apoptosis in all of the cell lines. Multivariate linear regression analyses revealed that the prediction improved when the intracellular Pt content or the Comet data were included in the analysis, for all sensitive cell lines and for the A2780 and A2780cis cell lines, respectively. Thus, a reliable cisplatin resistance predictive model, which combines the quantitation of adducts by HPLC-ICP-MS, and their repair, with the intracellular Pt content and induced genomic instability, might be essential to identify early therapy failure.
顺铂是癌症治疗中使用最广泛的金属药物之一，存在患者耐药这一重要缺陷。这种耐药性是由不同过程导致的，包括那些阻止DNA加合物形成和/或使其快速清除的过程。因此，一种准确检测和定量顺铂诱导的加合物的工具对于预测患者耐药性可能具有重要价值。为了证明这一假设的有效性，应用了高灵敏度等离子体质谱（ICP-MS）策略来测定DNA加合物水平和细胞内铂浓度。将这两个与金属相关的参数与对四种不同来源和顺铂敏感性的人类细胞系（A549、GM04312、A2780和A2780cis）的基因组稳定性（通过彗星试验）和细胞周期进程（通过流式细胞术）方面的生物学反应评估相结合，这些细胞系用低剂量顺铂（5、10和20μM，处理3小时）进行处理。将细胞活力和凋亡作为耐药指标进行测定。单变量线性回归分析表明，处理后1小时测量的顺铂诱导的G-G链内加合物定量是所有细胞系中活力和凋亡的最佳预测指标。多变量线性回归分析显示，分别对于所有敏感细胞系以及A2780和A2780cis细胞系，当分析中纳入细胞内铂含量或彗星试验数据时，预测效果得到改善。因此，一个可靠的顺铂耐药预测模型，该模型将通过HPLC-ICP-MS对加合物的定量及其修复与细胞内铂含量和诱导的基因组不稳定性相结合，对于识别早期治疗失败可能至关重要。
