Screening for von-Hippel Lindau gene mutations in chemically-induced kidney tumors of rats using intron-derived primers for PCR-SSCP analysis

von Hippel-Lindau (VHL) gene mutations occur throughout 3 exons including the exon-intron boundaries in human familial and sporadic renal cell carcinomas. To explore the possible role of the VHL gene in chemically-induced rat kidney tumors originating from various cell types, more than 150 bp of rat VHL intron sequences flanking the 3 exons were determined by dideoxy sequencing. Five primer sets were selected for PCR to amplify the coding regions of VHL exons 1-3 and the exon-intron boundaries. Rat kidney tumors (10 eosinophilic epithelial tumors; 10 nephroblastomas; 7 mesenchymal tumors) induced by various nitroso compounds were examined for VHL mutations by PCR-SSCP analysis. No mutation was detected in any tumor type, indicating that VHL mutations do not drive pathogenesis in rat kidney tumors arising from the distal region of the renal tubule, the metanephrogenic blastema, or stromal tissues of the cortex. This methodology may, however, be useful for screening other types of rat tumors for VHL mutations.

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