Use of a Low-Density Microarray for Studying Gene Expression Patterns Induced by Hepatotoxicants on Primary Cultures of Rat Hepatocytes

• Published in: Toxicological sciences 2003-10, Vol.75 (2), p.378-392
• Main Authors: de Longueville, Francoise, Atienzar, Franck A., Marcq, Laurence, Dufrane, Simon, Evrard, Stéphanie, Wouters, Lydia, Leroux, Florence, Bertholet, Vincent, Gerin, Brigitte, Whomsley, Rhys, Arnould, Thierry, Remacle, José, Canning, Mickael
• Format: Article
• Language: English
• Subjects: Animals; Biological and medical sciences; Chemical and industrial products toxicology. Toxic occupational diseases; Cytochrome P-450 Enzyme System - biosynthesis; Cytochrome P-450 Enzyme System - genetics; DNA microarrays; Dose-Response Relationship, Drug; drug metabolism; Drug toxicity and drugs side effects treatment; Enzyme Induction - drug effects; Formazans - metabolism; gene expression pattern; Gene Expression Profiling; Hepatocytes - drug effects; Hepatocytes - metabolism; hepatotoxicants; low-density microarray; Medical sciences; Oligonucleotide Array Sequence Analysis - methods; Pharmacology. Drug treatments; Rats; Rats, Wistar; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger - metabolism; Tetrazolium Salts - metabolism; Toxicity: digestive system; toxicogenomics; Toxicology; Various organic compounds; Xenobiotics - classification; Xenobiotics - toxicity
• ISSN: 1096-6080; 1096-0929; 1096-0929
• DOI: 10.1093/toxsci/kfg196

In the field of gene expression analysis, DNA microarray technology is having a major impact on many different areas including toxicology. For instance, a number of studies have shown that transcription profiling can generate the information needed to assign a compound to a mode-of-action class. In this study, we investigated whether compounds inducing similar toxicological endpoints produce similar changes in gene expression. In vitro primary rat hepatocytes were exposed to 11 different hepatotoxicants: acetaminophen, amiodarone, clofibrate, erythromycin estolate, isoniazid, α-naphtylylisothiocyanate, β-naphtoflavone, 4-pentenoic acid, phenobarbital, tetracycline, and zileuton. These molecules were selected on the basis of their variety of hepatocellular effects observed such as necrosis, cholestasis, steatosis, and induction of CYP P450 enzymes. We used a low-density DNA microarray containing 59 genes chosen as relevant toxic and metabolic markers. The in vitro gene expression data generated in this study were generally in good agreement with the literature, which mainly concerns in vivo data. Furthermore, gene expression profiles observed in this study have been confirmed for several genes by real-time PCR assays. All the tested drugs generated a specific gene expression profile. Our results show that even with a relatively limited gene set, gene expression profiling allows a certain degree of classification of compounds with similar hepatocellular toxicities such as cholestasis, necrosis. The clustering analysis revealed that the compounds known to cause steatosis were linked, suggesting that they functionally regulate similar genes and possibly act through the same mechanisms of action. On the other hand, the drugs inducing necrosis and cholestasis were pooled in the same cluster. The drugs arbitrarily classified as the CYP450 inducers formed individual clusters. In conclusion, this study suggests that low-density microarrays could be useful in toxicological studies.