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| Status |
Public on Apr 15, 2015 |
| Title |
Integration of metabolic activation with a predictive toxicogenomics signature to classify genotoxic versus nongenotoxic chemicals in human TK6 cells |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by array
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| Summary |
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 paper, a genomic biomarker was developed in human TK6 cells to classify chemicals as genotoxic or non-genotoxic. 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 non-genotoxic (dexamethasone, DEX; phenobarbital, PB) agents correctly was evaluated. Cells were exposed to increasing chemical concentrations for 4h and collected 0h, 4h and 20h post-exposure. Relative survival, apoptosis, and micronucleus frequency were measured at 24h. 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 non-genotoxic at all concentrations and time points. The high concentration of PB was misclassified at 24h, 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.
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| Overall design |
This experiment examined the whole genome transcriptional response of TK6 cells exposed to DMSO (+S9) and 10 μg/ml BaP (-S9) (both compared to DMSO (-S9)) for 4 hours followed by a 20h in fresh media (24h time point), in addition to 7.5 mM dexamethasone (+S9) (compared to DMSO (+S9)). Each condition had 3 biological replicates, which were pooled for gene expression analysis. There were a total 3 samples (6 arrays) included in the final analysis using a two-colour dye swap design.
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| Contributor(s) |
Buick JK, Moffat I, Swartz CD, Recio L, Williams A, Hyduke DR, Li H, Fornace AJ, Aubrecht J, Yauk CL |
| Citation(s) |
25733247 |
| Submission date |
Jun 24, 2014 |
| Last update date |
Oct 11, 2016 |
| Contact name |
Julie Buick |
| E-mail(s) |
julie.buick@canada.ca
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| Phone |
613-960-1599
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| Organization name |
Health Canada
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| Department |
Mechanistic Studies Division
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| Lab |
Genomics Lab
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| Street address |
50 Colombine Driveway
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| City |
Ottawa |
| State/province |
Ontario |
| ZIP/Postal code |
K1A 0K9 |
| Country |
Canada |
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| Platforms (1) |
| GPL14550 |
Agilent-028004 SurePrint G3 Human GE 8x60K Microarray (Probe Name Version) |
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| Samples (3) |
| GSM1419482 |
24h - Vehicle Control (+S9) - Pooled Replicate |
| GSM1419483 |
24h - Benzo[a]pyrene - 10 μg/ml (-S9) - Pooled Replicate |
| GSM1419484 |
24h - Dexamethasone - 7.5 mM (+S9) - Pooled Replicate |
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| Relations |
| BioProject |
PRJNA255257 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE58782_RAW.tar |
36.5 Mb |
(http)(custom) |
TAR (of TXT) |
| Processed data included within Sample table |
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