Application of the TGx-28.65 transcriptomic biomarker to classify genotoxic and non-genotoxic chemicals in human TK6 cells in the presence of rat liver S9.
Application of the TGx-28.65 transcriptomic biomarker to classify genotoxic and non-genotoxic chemicals in human TK6 cells in the presence of rat liver S9.
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DOI：
10.1002/em.22004
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发表时间：
2016-05
影响因子：
2.8
通讯作者：
Aubrecht, Jiri
中科院分区：
文献类型：
--
作者：
Yauk, Carole L.;Buick, Julie K.;Williams, Andrew;Swartz, Carol D.;Recio, Leslie;Li, Heng-Hong;Fornace, Albert J., Jr.;Thomson, Errol M.;Aubrecht, Jiri
关键词：
预测毒性的体外转录特征可以促进化学筛选。我们之前开发了一种转录组学生物标志物（称为TGx-28.65），用于将人类淋巴母细胞TK 6细胞中的药物分类为遗传毒性（DNA损伤）和非遗传毒性。由于TK 6细胞不表达细胞色素P450，我们证实了在共暴露于1% 5，6苯并噻二酮/苯巴比妥诱导的大鼠肝脏S9代谢活化的细胞中通过生物标志物进行的准确分类。然而，化学品可能需要不同类型的S9来激活。在此，我们研究了TK 6细胞对较高百分比的多氯联苯、苯并呋喃酮/苯巴比妥或乙醇诱导的大鼠肝脏S9的反应，以扩大TGx-28.65生物标志物的适用性。将TK 6细胞共暴露于供试化学品和S9 4 h后3 - 4 h，得出转录谱。初步研究证实，单独使用10%多氯联苯和5%乙醇诱导的S9不会诱导TGx-28.65生物标志物基因。然后使用其中一种S9（根据细胞存活率和微核诱导选择）检测7种遗传毒性和2种非遗传毒性化学品（以及并行溶剂和阳性对照）。在暴露后20小时，通过流式细胞术评估细胞的相对存活率和微核频率。使用不同的S9对遗传毒性/非遗传毒性化学品进行准确分类。用地塞米松和10%多氯联苯诱导的S9共处理的细胞的一个技术重复样本被错误地归类为遗传毒性，表明应谨慎使用高浓度的S9。即使是低浓度的遗传毒性化学品（不引起细胞毒性的化学品）也被正确分类，证明TGx-28.65是遗传毒性的敏感生物标志物。对13种化学品的数据集进行的Meta分析支持不同的S9可用于TK 6细胞，而不会损害使用TGx-28.65生物标志物的分类。Environ.摩尔变异体57：243-260，2016.© 2016 Her Majesty the Queen in Right of Canada.环境和分子诱变© 2016环境诱变剂协会
In vitro transcriptional signatures that predict toxicities can facilitate chemical screening. We previously developed a transcriptomic biomarker (known as TGx‐28.65) for classifying agents as genotoxic (DNA damaging) and non‐genotoxic in human lymphoblastoid TK6 cells. Because TK6 cells do not express cytochrome P450s, we confirmed accurate classification by the biomarker in cells co‐exposed to 1% 5,6 benzoflavone/phenobarbital‐induced rat liver S9 for metabolic activation. However, chemicals may require different types of S9 for activation. Here we investigated the response of TK6 cells to higher percentages of Aroclor‐, benzoflavone/phenobarbital‐, or ethanol‐induced rat liver S9 to expand TGx‐28.65 biomarker applicability. Transcriptional profiles were derived 3 to 4 hr following a 4 hr co‐exposure of TK6 cells to test chemicals and S9. Preliminary studies established that 10% Aroclor‐ and 5% ethanol‐induced S9 alone did not induce the TGx‐28.65 biomarker genes. Seven genotoxic and two non‐genotoxic chemicals (and concurrent solvent and positive controls) were then tested with one of the S9s (selected based on cell survival and micronucleus induction). Relative survival and micronucleus frequency was assessed by flow cytometry in cells 20 hr post‐exposure. Genotoxic/non‐genotoxic chemicals were accurately classified using the different S9s. One technical replicate of cells co‐treated with dexamethasone and 10% Aroclor‐induced S9 was falsely classified as genotoxic, suggesting caution in using high S9 concentrations. Even low concentrations of genotoxic chemicals (those not causing cytotoxicity) were correctly classified, demonstrating that TGx‐28.65 is a sensitive biomarker of genotoxicity. A meta‐analysis of datasets from 13 chemicals supports that different S9s can be used in TK6 cells, without impairing classification using the TGx‐28.65 biomarker. Environ. Mol. Mutagen. 57:243–260, 2016. © 2016 Her Majesty the Queen in Right of Canada. Environmental and Molecular Mutagenesis © 2016 Environmental Mutagen Society
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影响因子：
3.8
作者：
Ellinger-Ziegelbauer, Heidrun;Fostel, Jennifer M.;Aubrechtk, Jiri
通讯作者：
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影响因子：
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影响因子：
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作者：
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作者：
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Heflich, Robert H.
DOI：
10.1016/j.mrfmmm.2004.01.009
发表时间：
2004-05-15
影响因子：
2.3
作者：
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通讯作者：
Aubrecht, J