title: Comprehensive mutational analysis of the VHL gene in sporadic renal cell carcinoma: relationship to clinicopathological parameters.
abstract: To delineate more precisely the somatic von Hippel-Lindau disease (VHL) gene alteration as well as to elucidate its etiologic role in renal tumorigenesis, we examined a total of 240 sporadic renal cell carcinomas (RCCs) for somatic VHL gene alterations by DNA-SSCP followed by sequencing, methylation-specific PCR assay, microsatellite LOH study, and Southern blot analysis. Intragenic mutation of the VHL gene was found exclusively in clear-cell or variant-type RCCs at a frequency of 51% (104/202). Hypermethylation of the VHL promoter region was detected in an additional 11 clear-cell RCCs. Microsatellite analysis demonstrated that LOH of the VHL locus was found in 140/155 (90%) informative clear-cell RCCs. The VHL gene therefore seems to be inactivated in a two-hit manner by intragenic mutation or hypermethylation plus allelic loss in clear-cell RCC. Genomic rearrangement of the VHL gene detected by Southern analysis was not found (0/216 cases); this is in contrast to germ lines in which Southern aberrations consisted of 7-19% of the mutations. Clinicopathologic data demonstrated that VHL mutation/LOH did not vary according to tumor progression in clear-cell RCC, including tumor diameter, stage, grading, distant metastasis, and lymph node metastasis. Interestingly, VHL mutation was significantly less frequent in RCCs occurring in younger (< or = 55 years) than that in older (> or = 56 years) patients. These data suggested that the inactivation of the VHL tumor-suppressor gene is a specific genetic change in clear-cell RCC, and that it may occur at an early or first step in the clear-cell tumorigenic pathway rather than as a late event.
authors.affiliation: Department of Urology, Yokohama City University School of Medicine, Yokohama, Japan.
authors.forename: Keiichi
authors.initials: K
authors.lastname: Kondo
authors.forename: Masahiro
authors.initials: M
authors.lastname: Yao
authors.forename: Minoru
authors.initials: M
authors.lastname: Yoshida
authors.forename: Takeshi
authors.initials: T
authors.lastname: Kishida
authors.forename: Taro
authors.initials: T
authors.lastname: Shuin
authors.forename: Takeshi
authors.initials: T
authors.lastname: Miura
authors.forename: Masatoshi
authors.initials: M
authors.lastname: Moriyama
authors.forename: Kazuki
authors.initials: K
authors.lastname: Kobayashi
authors.forename: Naoki
authors.initials: N
authors.lastname: Sakai
authors.forename: Shigeki
authors.initials: S
authors.lastname: Kaneko
authors.forename: Satoshi
authors.initials: S
authors.lastname: Kawakami
authors.forename: Masaya
authors.initials: M
authors.lastname: Baba
authors.forename: Noboru
authors.initials: N
authors.lastname: Nakaigawa
authors.forename: Yoji
authors.initials: Y
authors.lastname: Nagashima
authors.forename: Yukio
authors.initials: Y
authors.lastname: Nakatani
authors.forename: Masahiko
authors.initials: M
authors.lastname: Hosaka
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000595:Amino Acid Sequence; D015139:Blotting, Southern; D002292:Carcinoma, Renal Cell / Q000235:genetics* / Q000473:pathology*; D019175:DNA Methylation; D004252:DNA Mutational Analysis; D004273:DNA, Neoplasm / Q000235:genetics; D005260:Female; D015321:Gene Rearrangement / Q000235:genetics; D006801:Humans; D007680:Kidney Neoplasms / Q000235:genetics* / Q000473:pathology*; D008025:Ligases / Q000235:genetics*; D019656:Loss of Heterozygosity / Q000235:genetics; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics; D025521:Tumor Suppressor Proteins; D044767:Ubiquitin-Protein Ligases; D051794:Von Hippel-Lindau Tumor Suppressor Protein
chemical_list: D004273:DNA, Neoplasm; D025521:Tumor Suppressor Proteins; D044767:Ubiquitin-Protein Ligases; D051794:Von Hippel-Lindau Tumor Suppressor Protein; D008025:Ligases; C497446:VHL protein, human