Application of a yeast assay to detect functional p53 mutations in archival prostate cancer tissue.
Detection and functional evaluation of mutant p53 alleles using a yeast assay could yield significant information for predicting the prognosis of patients with prostate cancer (CaP). Since the current version of this yeast assay is not applicable to archival tissues, we developed a modified assay for use on formalin-fixed, paraffin-embedded tissue and have applied it to the study of patient samples. Using this modified assay, we examined archival CaP samples from 10 patients for mutations in exons 5-8 of p53 gene. Mutations were detected in four samples: three resulted in the formation of red yeast colonies indicating complete loss of function, while one gave pink yeast colonies, indicating that this mutant retained partial function. In parallel, we analyzed these samples for p53 abnormalities using a single-strand conformational polymorphism (SSCP) approach. Only three of the four yeast-positive samples gave abnormal SSCP bands. In each case where abnormal p53 was found by both methods, DNA sequencing revealed the identical base change. These results suggest that the modified yeast assay may be more sensitive than SSCP for detection of p53 mutations, and demonstrate that the modified method can be used to detect and evaluate the function of p53 mutants present in archival tissue.
Shi
Xu-Bao
XB
Department of Urology, University of California, Davis School of Medicine, 4860 Y Street, Suite 3500, Sacramento, CA 95817, USA.
Di Mauro
Stephen M
SM
Highshaw
Ralph
R
Deitch
Arline D
AD
Evans
Christopher P
CP
Gumerlock
Paul H
PH
deVere White
Ralph W
RW
D016158:Genes, p53; D006801:Humans; D008297:Male; D009154:Mutation; D018807:Polymorphism, Single-Stranded Conformational; D011471:Prostatic Neoplasms / Q000235:genetics*; D015003:Yeasts / Q000235:genetics