Characterization and Interlaboratory Comparison of a Gene Expression Signature for Differentiating Genotoxic Mechanisms
Characterization and Interlaboratory Comparison of a Gene Expression Signature for Differentiating Genotoxic Mechanisms
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DOI：
10.1093/toxsci/kfp103
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发表时间：
2009-08-01
影响因子：
3.8
通讯作者：
Aubrechtk, Jiri
中科院分区：
文献类型：
--
作者：
Ellinger-Ziegelbauer, Heidrun;Fostel, Jennifer M.;Aubrechtk, Jiri
遗传毒性试验组合对化学致癌物的检测具有高度灵敏度。然而，它的特点是特异性低，只提供有限的机械信息所需的风险评估的积极结果。这在体外染色体损伤试验中出现阳性结果的情况下尤其重要，因为染色体损伤也可能继发于细胞死亡。越来越多的证据表明，细胞应激反应的毒理基因组学分析提供了一个深入了解遗传毒物的作用机制。为了评估这种毒理基因组学分析的效用，我们评估了TK6细胞的基因表达谱与四个模型的遗传毒性剂使用有针对性的高密度实时PCR方法在多实验室项目协调的健康与环境科学研究所委员会的基因组学在机制为基础的风险评估的应用。我们表明，这种基因分析技术在实验室中产生了可重复的数据，使我们能够得出结论，相关基因集的表达分析能够区分导致DNA加合物或双链断裂的化合物与干扰有丝分裂纺锤体功能或导致染色体损伤的化合物。此外，我们的数据表明，在早期时间点的基因表达谱是最有可能提供相关的遗传毒性损伤机制的信息，较大的基因表达阵列可能会提供更丰富的信息，用于区分遗传毒物的分子作用机制。虽然需要测试更多的化合物来识别稳健的分子特征，但本研究证实了毒理基因组学分析用于研究遗传毒性机制的潜力。
The genotoxicity testing battery is highly sensitive for detection of chemical carcinogens. However, it features a low specificity and provides only limited mechanistic information required for risk assessment of positive findings. This is especially important in case of positive findings in the in vitro chromosome damage assays, because chromosome damage may be also induced secondarily to cell death. An increasing body of evidence indicates that toxicogenomic analysis of cellular stress responses provides an insight into mechanisms of action of genotoxicants. To evaluate the utility of such a toxicogenomic analysis we evaluated gene expression profiles of TK6 cells treated with four model genotoxic agents using a targeted high density real-time PCR approach in a multilaboratory project coordinated by the Health and Environmental Sciences Institute Committee on the Application of Genomics in Mechanism-based Risk Assessment. We show that this gene profiling technology produced reproducible data across laboratories allowing us to conclude that expression analysis of a relevant gene set is capable of distinguishing compounds that cause DNA adducts or double strand breaks from those that interfere with mitotic spindle function or that cause chromosome damage as a consequence of cytotoxicity. Furthermore, our data suggest that the gene expression profiles at early time points are most likely to provide information relevant to mechanisms of genotoxic damage and that larger gene expression arrays will likely provide richer information for differentiating molecular mechanisms of action of genotoxicants. Although more compounds need to be tested to identify a robust molecular signature, this study confirms the potential of toxicogenomic analysis for investigation of genotoxic mechanisms.