Background: The aim of the study was to identify p53 gene mutations by FAMA (fluorescence-assisted mismatchanalysis) in colorectal cancers.Patients and methods: Analytical scanning of the p53 gene (exons 5–9) was performed in colon cancer samplesfrom 44 consecutive patients by FAMA. FAMA is a semiautomatic scanning approach based on the chemical cleavageof the mismatch in fluorescently labeled heteroduplex DNA, obtained from the combination of a normal and a mutatedallele. FAMA has already shown optimal levels of diagnostic accuracy and sensitivity in detecting gene mutations(nucleotide substitutions, insertions/deletions) both at the germline and somatic level. The peculiar feature of FAMA isits ability to detect and localize mutations, by a redundant pattern of signals due to fluorescent DNA fragmentsgenerated by chemical cleavage. Moreover, previous data have demonstrated that normal contaminating DNA fromstromal cells in the sample does not affect the sensitivity of the procedure, leading to the identification of the mutationeven when the ratio mutant/normal allele is 10%.Results: Eighteen mutations (12 missense, one nonsense, two deletions, three nucleotide substitutions at the level ofthe splice-junctions) and two polymorphisms were detected by FAMA in 17 patients (39%) and then confirmed byautomated sequence analysis. Six of 18 mutations (33%) were not previously reported for colon cancer samples andtwo of 18 lesions (11%) were identified as novel p53 mutations.Conclusions: Analytical scanning of the p53 gene by FAMA in DNA from colon cancer samples provides a sensitive,accurate and specific diagnostic procedure for routine clinical application.
|Numero di pagine||6|
|Rivista||Annals of Oncology|
|Stato di pubblicazione||Published - 2006|
All Science Journal Classification (ASJC) codes