PATHOLOGY & ONCOLOGY RESEARCHVol. 6 No. 4, 2000

 Article

p53 Protein Accumulation and p53 Gene Mutation in Colorectal Cancer

Anna NASIEROWSKA-GUTTMEJER1, Lech TRZECIAK3, Marek P NOWACKI2, Jerzy OSTROWSKI3

1Department of Pathology, Medical Center for Postgraduate Education, and Maria Sk³odowska-Curie Memorial Cancer Center and Institute of Oncology, Warsaw, Poland
2Department of Colorectal Diseases, the Maria Sk³odowska-Curie Memorial Cancer Center and Institute of Oncology, Warsaw, Poland
3Department of Gastroenterology, Medical Center for Postgraduate Education and Maria Sk³odowska-Curie Memorial Cancer Center and Institute of Oncology, Warsaw, Poland

 

Comparison of immunohistochemical methods for detection of protein p53 accumulation and molecular techniques for analysis p53 gene mutation in colorectal cancer is presented. Thirty eight patients were included: all underwent surgery without preoperative treatment. Sex of patients, tumor localisa-tion, macro and microscopic type of cancer and staging according to Astler-Coller and Jass classifications were evaluated. Protein p53 accumulation was detected by the streptavidin-biotin method using DO-7 (Dako) antibody. The number of cells stained were classified semiquanititatively according to a scoring system: (–)no positive cells, (+) : 10-30% positive cells, (++) : 40-70% positive cells, (+++) : >70% positive cells. For all cancer samples, exons 5 to 9 of p53 gene were amplified from isolated genomic DNA. PCR products were subjected to single standed conformational polymorphism analysis. All product were also directly sequenced on ABI PRISM 377 apparatus using fluorescent dideoxyterminators chemistry. The protein p53 accumulation was detected in 53% (20/38), whereas p53 gene mutation was seen in 55% (21/38). Among them, 15 patients (39%) with overexpression showed mutation in exon 5-8 gene p53. Discrepancies between results were noted in 29%. In conclusion, the necessity of both methods – immunohistochemical and molecular – is indicated for the objective evaluation of functional and structural status of p53 gene and protein. Pathology & Oncology Research, Vol 6, Nr 4, 275-279, 2000

Key words: p53 protein; p53 gene; molecular biology; immunohistochemical studies; colorectal cancer


Received: May 18, 2000; accepted: Nov 28, 2000
Correspondence: Anna NASIEROWSKA-GUTTMEJER, Department of Pathology, Medical Center for Postgraduate Education, and Maria Sk³odowska-Curie Memorial Cancer Center and Institute of Oncology, Roentgena str. 5 Warsaw 02-971, Poland, Fax: 022-6440208

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