Integration of metabolic activation with a predictive toxicogenomics signature to classify genotoxic versus nongenotoxic chemicals in human TK6 cells
Integration of metabolic activation with a predictive toxicogenomics signature to classify genotoxic versus nongenotoxic chemicals in human TK6 cells
复制标题
DOI：
10.1002/em.21940
复制
发表时间：
2015-07-01
影响因子：
2.8
通讯作者：
Yauk, Carole L.
中科院分区：
文献类型：
--
作者：
Buick, Julie K.;Moffat, Ivy;Yauk, Carole L.
在遗传毒理学中使用综合方法，包括结合基因表达数据来确定参与反应的分子途径，正变得越来越普遍。在一篇相关文章中，在人类TK6细胞中开发了一种基因组生物标志物，用于将化学物质分类为遗传毒性或非遗传毒性。由于TK6细胞不具有代谢能力，我们开始扩大该生物标志物的应用范围，用于需要代谢激活的化学物质。具体来说，化学暴露是在大鼠肝脏S9存在的情况下进行的。评估生物标志物对遗传毒性（苯并[a]芘，BaP；黄曲霉毒素B1， AFB1）和非遗传毒性（地塞米松，DEX；苯巴比妥，PB）药物的正确分类能力。细胞暴露于浓度增加的化学物质中4小时，并在暴露后0小时、4小时和20小时收集。在24小时测量相对存活率、细胞凋亡和微核频率。用安捷伦微阵列测量转录组谱。统计建模和生物信息学工具应用基因组生物标志物对每种化学物质进行分类。在三个时间点上，BaP和AFB1均被正确归类为中、高浓度遗传毒性，而DEX在所有浓度和时间点上均被正确归类为非遗传毒性。高浓度PB在24小时出现误分类，提示较晚时间点的细胞毒性可能导致误分类。这些数据表明，使用S9不会损害生物标志物对TK6细胞遗传毒性进行分类的能力。最后，我们证明了生物标志物也能够使用来自人类HepaRG细胞的公开数据集准确分类遗传毒性。环绕。中国生物医学工程学报，2015,36(6):532 -534。(c) 2015年作者。环境与分子诱变出版于Wiley期刊公司。
The use of integrated approaches in genetic toxicology, including the incorporation of gene expression data to determine the molecular pathways involved in the response, is becoming more common. In a companion article, a genomic biomarker was developed in human TK6 cells to classify chemicals as genotoxic or nongenotoxic. Because TK6 cells are not metabolically competent, we set out to broaden the utility of the biomarker for use with chemicals requiring metabolic activation. Specifically, chemical exposures were conducted in the presence of rat liver S9. The ability of the biomarker to classify genotoxic (benzo[a]pyrene, BaP; aflatoxin B1, AFB1) and nongenotoxic (dexamethasone, DEX; phenobarbital, PB) agents correctly was evaluated. Cells were exposed to increasing chemical concentrations for 4 hr and collected 0 hr, 4 hr, and 20 hr postexposure. Relative survival, apoptosis, and micronucleus frequency were measured at 24 hr. Transcriptome profiles were measured with Agilent microarrays. Statistical modeling and bioinformatics tools were applied to classify each chemical using the genomic biomarker. BaP and AFB1 were correctly classified as genotoxic at the mid- and high concentrations at all three time points, whereas DEX was correctly classified as nongenotoxic at all concentrations and time points. The high concentration of PB was misclassified at 24 hr, suggesting that cytotoxicity at later time points may cause misclassification. The data suggest that the use of S9 does not impair the ability of the biomarker to classify genotoxicity in TK6 cells. Finally, we demonstrate that the biomarker is also able to accurately classify genotoxicity using a publicly available dataset derived from human HepaRG cells. Environ. Mol. Mutagen. 56:520-534, 2015. (c) 2015 The Authors. Environmental and Molecular Mutagenesis Published by Wiley Periodicals, Inc.