Yeast functional assay of the p53 gene status in human cell lines maintained in our laboratory.
We used a yeast functional assay (functional analysis of separated alleles in yeast: FASAY) to determine the p53 gene status of human cell lines maintained in our laboratory. This assay enables the researcher to score wild-type p53 expression on the basis of the ability of expressed p53 to transactivate the reporter gene HIS 3 via the p53-responsive GAL 1 promoter in Saccharomyces cerevisiae. The cell lines examined were ten hepatoma, two hepatoblastoma, three in vitro immortalized fibroblast, two osteosarcoma, a chondrosarcoma, an ovarian teratocarcinoma and a colon cancer cell line. Out of 20 cell lines, 11 cell lines had mutations in both alleles of the p53 gene, and another 8 cell lines had no mutation in the p53 gene. Thus, 55% of the cell lines examined had mutations in the p53. Interestingly, PA-1 cells had both the normal and the mutant p53 alleles, showing that FASAY is a useful method for detecting the wild-type and mutated p53 genes simultaneously. As for the three liver cell lines harboring HBsAg, there was no relationship between their p53 gene status and the presence of HBsAg. Two cell lines were normal for p53 status, while the other had a mutation of the p53 gene.
Mihara
K
K
Department of Cell Biology, Okayama University Medical School, Japan.
Miyazaki
M
M
Kondo
T
T
Fushimi
K
K
Tsuji
T
T
Inoue
Y
Y
Fukaya
K
K
Ishioka
C
C
Namba
M
M
D000483:Alleles; D006528:Carcinoma, Hepatocellular / Q000235:genetics; D016158:Genes, p53; D015316:Genetic Therapy; D006801:Humans; D008113:Liver Neoplasms / Q000235:genetics; D009154:Mutation; D016133:Polymerase Chain Reaction; D014407:Tumor Cells, Cultured; D015003:Yeasts / Q000235:genetics*