Detection of p53 point mutations in primary human vulvar cancer by PCR and temperature gradient gel electrophoresis.
Clinical and experimental evidence is consistent with a key role of point mutations of the p53 tumor suppressor gene in the etiology of squamous cell carcinoma. To determine the relation of tumor behavior and patient survival in vulvar cancer in regard to p53 status, we retrospectively analyzed 38 paraffin-embedded specimens of primary vulvar cancer for genetic alterations of exons 5-8 of the p53 gene. For detection of p53 point mutations we used in vitro amplification by polymerase chain reaction (PCR) and temperature gradient gel electrophoresis (TGGE) and, as a detection method, direct sequencing for mutation verification. p53 point mutations were detected in 12/38 tumor specimens. Patients bearing p53 point mutations showed a significantly shorter relapse-free (log-rank test, P = 0.002) and overall survival time (log-rank test, P = 0.0006). We conclude that PCR-TGGE is an appropriate method for detection of p53 point mutations in paraffin-embedded material. We show that loss of wild-type p53 is an adverse prognostic factor in patients suffering from vulvar cancer.
Sliutz
G
G
Department of Gynecology and Obstetrics, General Hospital, University of Vienna, Austria.
Schmidt
W
W
Tempfer
C
C
Speiser
P
P
Gitsch
G
G
Eder
S
S
Schneeberger
C
C
Kainz
C
C
Zeillinger
R
R
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D001483:Base Sequence; D004252:DNA Mutational Analysis; D004587:Electrophoresis, Agar Gel; D005091:Exons; D005260:Female; D016158:Genes, p53 / Q000235:genetics*; D006358:Hot Temperature; D006801:Humans; D008875:Middle Aged; D008969:Molecular Sequence Data; D017354:Point Mutation; D016133:Polymerase Chain Reaction; D012189:Retrospective Studies; D014846:Vulvar Neoplasms / Q000235:genetics*