A comparative study of detection of p53 mutations in human breast cancer by flow cytometry, single-strand conformation polymorphism and genomic sequencing

Abstract

The accuracy of immunodetection by dual parameter flow cytometry (FCM), polymerase chain reaction-mediated single strand conformation polymorphism (PCR-SSCP) and genomic sequencing to detect p53 mutations were compared. Analysis by the last two techniques was restricted to exons 5-8. Initially, 110 breast tumours were screened for p53 expression by FCM. Seventy (64%) of tumours were immunopositive. Fifteen highly immunopositive and 15 completely immunonegative tumours were selected for further analysis by PCR-SSCP and genomic sequencing. Eleven out of 15 immunopositive tumours were found to have mutation by PCR-SSCP. Genomic sequencing confirmed the presence of mutation in 10 of these 11 immunopositive tumours. Therefore, four immunopositive tumours failed to show mutation by SSCP and five by genomic sequencing. Of the 15 immunonegative tumours, one showed mutation by both PCR-SSCP and genomic sequencing and one tumour has undergone deletion of the p53 gene. Overall, immunoreactivity correlated with both PCR-SSCP and genomic sequencing in 80% of cases (24/30), and there was 96.5% (28/29) concordance between PCR-SSCP and genomic sequencing. We conclude that there is good concordance between mutations detected by PCR-SSCP and genomic sequencing, but immunochemical detection of p53 overexpression is not an absolute indicator of p53 gene mutation.