Molecular diagnosis of von Hippel-Lindau disease (VHL) using VHL tumor suppressor gene and the research for the VHL gene product

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To define the molecular basis of VHL patients in Japanese populations, we collected 72 unrelated Japanese VHL patients and completely tested for germline mutations of the VHL gene in 45 VHL patients by single-strand conformation polymorphism (SSCP) analysis and Southern blot analysis. We detected 23 (51%) intragenic mutations of the VHL gene and three (6.7%) deletions by SSCP analysis and Southern blot respectively. The intragenic mutations consisted of 14 missense mutations, seven microdeletions or insertions and two splice-site mutations. Nine of 10 mutations in exon 1 are localized in a short region of 37 nucleotides. Five unique sites of mutation were included, which were not seen in prevoius stuidied. Unlike Western VHL patients, nonsense mutations were not found in Japaneses VHL patients. The mutations found in 22 VHL patients without pheochromocytoma consisted of 11 missense mutations, six microdeletions or insertions, two splice-site alterations and three deletions. The mutations found in four VHL patients with pheochromocytomas consisted of one missense mutation at nucleotide 683 (codon 228), two missense mutations at uncleotide 712 (codon 38) and a novel 20 bp insertion at nucleotide 776 (codon 259). Our results suggest that unique features compared with those in Western patients.In situ hybridyzation study with VHL gene probe showed expression of VHL gene at cerebral and cerebellar neuronal cells and stromal cells of hemangioblastoma. In addition, VHL protein expression with anti-VHL protein monoclonal antibody showed expression of VHL protein at neuronal cells and brain tumor cells.