title: P53 in breast carcinomas: association between presence of mutation and immunohistochemical expression using a semiquantitative approach.
abstract: The aim of this study was to evaluate the concordance between the presence of p53 mutations in breast carcinomas expressing the protein by immunohistochemistry. A series of 60 breast carcinomas was evaluated by immunohistochemistry using monoclonal antibodies against p53 protein (DO 7 and PAb 1801). Twenty cases classified as being positive for p53 according to the current approach (if 5% or more of neoplastic cells contained reaction product in the nucleus) were used for molecular studies. These cases were re-assessed semi-quantitatively using a scoring system based on intensity and percentage of stained cells. DNA was phenol-chloroform extracted from microdissected normal and tumour cells obtained from formalin-fixed, paraffin-embedded tissue sections. Mutations in the p53 gene were analysed by SSCP (single strand conformational polymorphism) with primers covering exons 2-3 to 11. Ten out of the 20 p53-positive cases presented mutations detected by SSCP analysis. Mutations have been found in several exons ranging from exon 4 to exon 10. We observed a positive relationship between the presence of mutations and immunohistochemical evaluation of p53 protein expression using a semiquantitative scoring system. All cases with more than 2/3 stained tumour cells and strong intensity of staining exhibited p53 mutations. At variance, no p53 mutations were found in cases with less than 1/3 stained tumour cells and moderate intensity of staining. Therefore, only the identification of positivity for p53 detected by immunohistochemistry did not always reflect the detection of p53 mutations in breast cancer, however the use of a semi-quantitative approach seems to be useful as an indicator of the presence of mutation.
authors.affiliation: Institute of Molecular Pathology and Immunology, IPATIMUP, Porto Medical School, Portugal. fernando.schmitt@ipatimup.pt
authors.forename: F C
authors.initials: FC
authors.lastname: Schmitt
authors.forename: R
authors.initials: R
authors.lastname: Soares
authors.forename: L
authors.initials: L
authors.lastname: Cirnes
authors.forename: R
authors.initials: R
authors.lastname: Seruca
mesh_terms: D000328:Adult; D000368:Aged; D001943:Breast Neoplasms / Q000737:chemistry / Q000235:genetics* / Q000473:pathology; D018270:Carcinoma, Ductal, Breast / Q000737:chemistry / Q000235:genetics* / Q000473:pathology; D004273:DNA, Neoplasm / Q000032:analysis; D005260:Female; D016158:Genes, p53 / Q000235:genetics*; D006801:Humans; D007124:Immunoenzyme Techniques; D008875:Middle Aged; D020125:Mutation, Missense; D016133:Polymerase Chain Reaction; D018807:Polymorphism, Single-Stranded Conformational; D016159:Tumor Suppressor Protein p53 / Q000032:analysis
chemical_list: D004273:DNA, Neoplasm; D016159:Tumor Suppressor Protein p53