Abstract
Ferric nitrilotriacetate induces oxidative damage in renal proximal tubules, a consequence of Fenton-like reaction, that ultimately leads to a high incidence of renal cell carcinoma (RCC) in rats. In order to find common genetic alterations in this oxystress-induced carcinogenesis model, RCCs were produced in F1 hybrid rats between Wistar and Long-Evans strains and genomes were screened for loss of heterozygosity (LOH) with microsatellite polymorphic markers by PCR. Five consecutive markers on chromosome 5 (D5Mgh5, D5Mit9, D5Mgh6, D5Mit11 and D5Mit6) showed LOH in >40% of the RCCs. As possible candidate tumor suppressor genes on chromosome 5, p15INK4B and p16INK4A were investigated for genetic alteration and aberrant methylation by Southern blot, PCR/SSCP/ sequencing and methylation-specific PCR. Genetic alteration (homozygous or hemizygous deletion with or without point mutation) or aberrant methylation were found in 30.7 and 53.8% of the RCC cases, respectively, which was proportionally associated with the histological nuclear grade and metastatic activity. Our data suggest that inactivation of p15 and p16 genes could be one of the major pathways responsible for oxystress-induced carcinogenesis.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Base Sequence
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Blotting, Southern
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Carcinogens / toxicity
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Carcinoma, Renal Cell / chemically induced
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Carcinoma, Renal Cell / genetics*
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Carcinoma, Renal Cell / pathology
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Carrier Proteins / genetics*
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Cell Cycle Proteins*
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Chromosomes, Human, Pair 5 / genetics*
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CpG Islands
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Crosses, Genetic
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Cyclin-Dependent Kinase Inhibitor p15
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Cyclin-Dependent Kinase Inhibitor p16*
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DNA Methylation
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DNA, Neoplasm / genetics
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Ferric Compounds / toxicity
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Gene Deletion
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Genes, Tumor Suppressor*
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Genes, p16*
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Humans
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Kidney Neoplasms / chemically induced
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Kidney Neoplasms / genetics*
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Kidney Neoplasms / pathology
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Kidney Tubules, Proximal / drug effects
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Loss of Heterozygosity
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Microsatellite Repeats
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Molecular Sequence Data
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Nitrilotriacetic Acid / analogs & derivatives
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Nitrilotriacetic Acid / toxicity
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Oxidative Stress
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Point Mutation
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Polymerase Chain Reaction
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Polymorphism, Single-Stranded Conformational
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Rats
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Rats, Wistar
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Tumor Suppressor Proteins*
Substances
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CDKN2B protein, human
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Carcinogens
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Carrier Proteins
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Cdkn2b protein, rat
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Cell Cycle Proteins
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Cyclin-Dependent Kinase Inhibitor p15
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Cyclin-Dependent Kinase Inhibitor p16
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DNA, Neoplasm
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Ferric Compounds
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Tumor Suppressor Proteins
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Nitrilotriacetic Acid
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ferric nitrilotriacetate