p53 immunoreactivity and single-strand conformational polymorphism analysis often fail to predict p53 mutational status

The intent of this study was to investigate the ability of p53 expression a nd single-strand conformational polymorphism analysis (SSCP) to predict p53 mutational status in archival, paraffin-embedded tissues of gastric cancer . We evaluated paraffin-embedded tissues from 78 patients with advanced gas tric cancer. The mutational status of the p53 gene (exons 5-9) was examined by SSCP analysis and by direct sequencing, These results were compared wit h p53 expression as assessed by immunohistochemical analysis (IHC). We grad ed p53 expression on a scale from 0 to 8 on the basis of both the intensity and the number of cells staining. Overall, we detected p53 immunoreactivit y in 75.6% of the gastric cases; 19 (32.2%) of these cases scored from 1 to 4, and 40 (67.8%) cases scored from 5 to 8, p53 gene mutations were detect ed in 18 cases (23.1%) by SSCP and in 28 cases (36%) by direct sequencing. Thus, SSCP failed to detect 38% of the mutations found by sequencing. The m ajority of missed mutations involved exons 7 and 8. The concordance between IHC and SSCP was 37%, and the concordance between IHC and direct sequencin g was 50%, Forty-five percent of cases positive by IHC failed to show mutat ions in exons 5 through 9. Five percent of cases negative by IHC (4 cases) contained mutations. One had a 1-base pair insertion; one had a mutation th at resulted in a stop codon; the third had a mutation in exon 8; and the fo urth had a mutation in both exons 5 and 8. Our findings indicate that p53 i mmunoreactivity correlates with the presence or absence of gene mutations i n 50% of advanced gastric cancers when exons 5 through 9 are examined and t hat IHC cannot be reproducibly used as a marker of mutation in the most com monly mutated exons of the p53 gene. Furthermore, the sensitivity of SSCP f or detecting mutations is only 62%, Thus, SSCP analysis cannot be used reli ably to screen for p53 mutations.