Molecular Detection of Von Hippel-Lindau Gene Mutations in Urine and Lymph Node Samples in Patients with Renal Cell Carcinoma: Potential Biomarkers for Early Diagnosis and Postoperative Metastatic Status

Abstract

These data indicate the presence of detectable levels of tumor derived DNA in the urine of patients with renal cell carcinoma and suggest that nested single strand conformational polymorphism analysis of VHL gene of urine samples provides a possible tool for the early detection of renal cell carcinoma. Furthermore, mutation specific nested reverse transcription polymerase chain reaction is useful to detect occult lymph node micrometastasis and may predict patients at risk for local recurrence. These 2 combined approaches using VHL gene mutations may contribute to the total therapy for and prognosis of renal cell carcinoma.

Associations between VHL genotype and clinical phenotype in familial von Hippel–Lindau disease

“…Phenotypic expression of VHL disease is deeply influenced by genetic modifiers [48] and the genotypes can now be determined easily using MLPA, qPCR with TaqMan probes and by LOH and SSCP analyses. Such multiple methods substantially improve our ability to link particular genotypes with clinical manifestations and can facilitate the clinical diagnosis of VHL disease [49,50]. In addition, using DNA testing, individuals with no symptoms have been found to have changes in the VHL gene regardless of age.…”

Germline mutations in the von Hippel–Lindau disease (VHL) gene in mainland Chinese families

“…Genomic DNA (50-150 ng) was ampliWed by PCR in 50 l reaction mixture containing 10£ buVer 5 l, 200 M dNTP 1 l, 10% DMSO 1.5 l, Taq Polymerase 1.0 U and 10 pmol/ l of each primer 2 l. PCR conditions were 40 cycles of 94°C for 45 s, 59°C for 45 s, and 72°C for 45 s. Primers used for PCR and sequencing were as same as previous reports (Ashida et al 2003;Ashida et al 2000). PCR products were puriWed and subjected to sequence analysis using an ABI 3730 automated sequencer.…”

Classical pathology versus molecular pathology in renal cell carcinoma

“…Different histologic subtypes other than conventional (clear cell) RCC have also been shown to harbor alterations in the VHL gene [21]. Further, VHL mutations can be detected in peripheral blood and urine [22,23]. Studies have shown a strong association between VHL gene alterations and advanced stage [24].…”