Correlation between p53 gene mutations and p53 protein accumulation evaluated by different methodologies.
Mutations in the p53 gene are the most common genetic alterations in many tumour histotypes. Many of these mutations induce conformational changes resulting in p53 protein stabilisation and consequently an accumulation detectable with immunochemical methods. Available data on the correlation between p53 gene alterations and p53 overexpression widely vary. In this study we analysed the correlation between p53 gene alterations detected by DGGE, SSCP and sequencing and protein expression detected by flow cytometric and immunohistochemical approaches by using PAb 1801 antibody. The study was performed on 21 bladder tumours and 10 cell lines derived from different tumour histotypes as representative of different methodologic problems which can be met starting from different types of biological material. The best correlation (81%) was observed between p53 mutations and FCM results, using a double evaluation criterion for the latter which includes the percentage of positive cells and "delta values", evaluated as the difference between the mean values of Pab 1801 stained cells and isotypic control. The high correlation obtained between results from this FCM double criterion and p53 gene mutations is a good starting point for the analysis on large series of tumours and for a multiparameter FCM analysis including p53 protein levels.
Calistri
D
D
Istituto Oncologico Romagnolo, Forlì, Italy.
Barzanti
F
F
Dal Susino
M
M
Fedriga
R
R
Saragoni
L
L
Bernardi
L
L
Ricotti
L
L
Zoli
W
W
D002295:Carcinoma, Transitional Cell / Q000737:chemistry / Q000235:genetics; D004252:DNA Mutational Analysis / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D005188:False Negative Reactions; D005434:Flow Cytometry; D005629:Frozen Sections; D016158:Genes, p53; D006801:Humans; D007124:Immunoenzyme Techniques; D019656:Loss of Heterozygosity; D009369:Neoplasms / Q000737:chemistry / Q000235:genetics* / Q000473:pathology; D018807:Polymorphism, Single-Stranded Conformational; D011487:Protein Conformation; D016159:Tumor Suppressor Protein p53 / Q000032:analysis*; D001749:Urinary Bladder Neoplasms / Q000737:chemistry / Q000235:genetics
D016159:Tumor Suppressor Protein p53