Assessment of cisplatin-induced nephrotoxicity by microarray technology.
Microarrays are a new technology used to study global gene expression and to decipher biological pathways. In the current study, microarrays were used to examine gene expression patterns associated with cisplatin-mediated nephrotoxicity. Sprague-Dawley rats received either single or seven daily ip doses of cisplatin (0.5 or 1 mg/kg/day) or the inactive isomer transplatin (1 or 3 mg/kg/day). Histopathological evaluation revealed renal proximal tubular necrosis in animals that received cisplatin for 7 days, but no hepatotoxic findings. Microarray analyses were performed using rat specific arrays containing 250 toxicity-related genes. Prominent gene expression changes were observed only in the kidneys of rats that received cisplatin for 7 days. Mechanistically, the gene expression pattern elicited by cisplatin (e.g., Bax upward arrow and SMP-30 downward arrow) suggested the occurrence of apoptosis and the perturbation of intracellular calcium homeostasis. The induction of multidrug resistance genes (MDR1 upward arrow, P-gp upward arrow) and tissue remodeling proteins (clusterin upward arrow, IGFBP-1 upward arrow, and TIMP-1 upward arrow) indicated the development of cisplatin resistance and tissue regeneration. Select gene expression changes were further confirmed by TaqMan analyses. Gene expression changes were not observed in the liver following cisplatin administration. In contrast to these in vivo findings, studies using NRK-52E kidney epithelial cells and clone-9 liver cells suggested that liver cells were more sensitive to cisplatin treatment. The discrepancies between the in vivo and in vitro results suggest that caution should be taken when extrapolating data from in vivo to in vitro systems. Nonetheless, the current study elucidates the biochemical pathways involved in cisplatin toxicity and demonstrates the utility of microarrays in toxicological studies.
Huang
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Department of Toxicology, Boehringer Ingelheim Pharmaceuticals, Inc., 900 Ridgebury Road, P.O. Box 368, Ridgefield, Connecticut 06877-0368, USA. qhuang@rdg.boeringer-ingelheim.com
Dunn
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Jayadev
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DiSorbo
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Pack
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Farr
S B
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Stoll
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Blanchard
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D000818:Animals; D002135:Calcium-Binding Proteins / Q000378:metabolism; D002460:Cell Line; D002945:Cisplatin / Q000008:administration & dosage / Q000633:toxicity*; D051152:Clusterin; D004847:Epithelial Cells / Q000187:drug effects / Q000378:metabolism; D015870:Gene Expression / Q000187:drug effects*; D019450:Genes, MDR / Q000187:drug effects; D006023:Glycoproteins / Q000378:metabolism; D022781:Hepatocytes / Q000187:drug effects; D007274:Injections, Intraperitoneal; D018970:Insulin-Like Growth Factor Binding Protein 1 / Q000378:metabolism; D007668:Kidney / Q000187:drug effects*; D007683:Kidney Tubular Necrosis, Acute / Q000139:chemically induced / Q000378:metabolism; D008099:Liver / Q000187:drug effects; D008297:Male; D018832:Molecular Chaperones / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis; D016133:Polymerase Chain Reaction; D011518:Proto-Oncogene Proteins / Q000378:metabolism; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism; D011897:Random Allocation; D051381:Rats; D017207:Rats, Sprague-Dawley; D013237:Stereoisomerism; D015238:Sulfotransferases; D013997:Time Factors; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000378:metabolism; D051028:bcl-2-Associated X Protein
C494828:Bax protein, rat; D002135:Calcium-Binding Proteins; D051152:Clusterin; D006023:Glycoproteins; D018970:Insulin-Like Growth Factor Binding Protein 1; D018832:Molecular Chaperones; D011518:Proto-Oncogene Proteins; D019253:Proto-Oncogene Proteins c-bcl-2; D019715:Tissue Inhibitor of Metalloproteinase-1; D051028:bcl-2-Associated X Protein; C034697:transplatin; D015238:Sulfotransferases; C028152:alcohol sulfotransferase; D002945:Cisplatin