ABSTRACT
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.
Subject(s)
Cisplatin/toxicity , Gene Expression/drug effects , Kidney/drug effects , Oligonucleotide Array Sequence Analysis , Animals , Calcium-Binding Proteins/metabolism , Cell Line , Cisplatin/administration & dosage , Clusterin , Epithelial Cells/drug effects , Epithelial Cells/metabolism , Genes, MDR/drug effects , Glycoproteins/metabolism , Hepatocytes/drug effects , Injections, Intraperitoneal , Insulin-Like Growth Factor Binding Protein 1/metabolism , Kidney Tubular Necrosis, Acute/chemically induced , Kidney Tubular Necrosis, Acute/metabolism , Liver/drug effects , Male , Molecular Chaperones/metabolism , Polymerase Chain Reaction , Proto-Oncogene Proteins/metabolism , Proto-Oncogene Proteins c-bcl-2/metabolism , Random Allocation , Rats , Rats, Sprague-Dawley , Stereoisomerism , Sulfotransferases , Time Factors , Tissue Inhibitor of Metalloproteinase-1/metabolism , bcl-2-Associated X ProteinABSTRACT
Heterozygous p53+/- transgenic mice are being studied for utility as a short-term alternative model to the 2-yr rodent carcinogenicity bioassay. During a 26-wk study to assess the potential carcinogenicity of oxymetholone using p-cresidine as a positive control, glass/polypropylene microchips (radio transponder identification devices) were subcutaneously implanted into male and female p53+/- mice. During week 15, the first palpable mass was clinically observed at an implant site. This rapidly growing mass virtually quadrupled in size by week 25. Microscopic examination of all implant sites revealed that 18 of 177 animals had a subcutaneous histologically malignant sarcoma. The neoplasms were characterized as undifferentiated sarcomas unrelated to drug treatment, as indicated by the relatively even distribution among dose groups, including controls. An unusual preneoplastic mesenchymal change characterized by the term "mesenchymal dysplasia" was present in most groups and was considered to be a prodromal change to sarcoma development. The tumors were observed to arise from dysplastic mesenchymal tissue that developed within the tissue capsule surrounding the transponder. The preneoplastic changes, including mesenchymal dysplasia, appeared to arise at the transponder's plastic anchoring barb and then progressed as a neoplasm to eventually surround the entire microchip. Capsule membrane endothelialization, inflammation, mesenchymal basophilia and dysplasia, and sarcoma were considered unequivocal preneoplastic/neoplastic responses to the transponder and were not related to treatment with either oxymetholone or p-cresidine.
Subject(s)
Genes, p53/genetics , Polypropylenes/adverse effects , Sarcoma, Experimental/pathology , Transducers/adverse effects , Anabolic Agents/toxicity , Animals , Carcinogens/toxicity , DNA, Neoplasm/analysis , DNA, Neoplasm/genetics , Female , Heterozygote , Male , Mice , Mice, Inbred C57BL , Mice, Transgenic , Oxymetholone/toxicity , Sarcoma, Experimental/etiology , Sarcoma, Experimental/genetics , Skin/drug effects , Skin/pathology , Skin/ultrastructure , Skin Neoplasms/etiology , Skin Neoplasms/genetics , Skin Neoplasms/pathology , Survival AnalysisABSTRACT
Light microscopic and electron microscopic findings in thymuses from 4-week old feline leukemia virus-infected and 4- and 9-week old noninfected kittens were evaluated and found to be morphologically similar to each other. Thymuses from 9-week old feline leukemia virusinfected kittens were markedly atrophied and individual lobules within each thymus varied in the severity of atrophy. Loubules having the least severe atrophy had a moderate thinning of the cortex and a heterogeneous thymuses included intense eosinopoiesis at the corticomedullary junction, increased prominence of vasculature, and enlarged Hassal's corpuscles. In addition to these changes lobules of thymus having the most severe atrophy had a marked cortical thymocyte depletion, lobule collapse, and increased numbers of mast cells. Degeneration of epithelial cells in most lobules was indicated by electronlucency of the cytoplasmic matrix and often greatly dilated rough endoplasmic reticulum.
