Immunohistochemical Study of p53 Protein in Colon, Gastric and Lung Cancer and Comparison with the Study of Genetic Alteration by the Method of Reverse Transcription Polymerase Chain Reaction Single Strand Conformation Polymorphisms

Masafumi Kataoka, Takahiro Okabayashi, Akira Nakashima, Shin Nakatani, Hiroki Johira, Kunzo Orita, Takato Noumi*, Hiroshi Kanazawa*

First Department of Surgery, Okayama University Medical School, *Okayama University Department of Biotechnology, Faculty of Engineering Science

Aberrations of p53 protein were investigated in surgical specimens of colon, gastric and lung cancer immunohistochemically, and to confirm the relation between immunohistochemical abnormality and genetic alteration, mutations in the DNA from specimens of colon cancer were analyzed by reverse transcriptionpolymerase chain reaction-single strand comformation polymorphism (RT-PCR-SSCP) method and subsequent direct sequence. Microwave-fixed paraffin-enbedded specimens were stained by monoclonal anbibody (PAb 1801, Oncogene Science Inc.) raised against p53 protein. Nine of 13 cases of colon cancer (69.2%), five of 8 cases of gastric cancer (62.5%) and four of 5 cases of lung cancer (80.0%) showed positive staining in the cancer cell nuclei. No significant correlation was found between immunohistochemical results and pathological findings. The RT-PCR-SSCP method detected mutations in six of 13 cases of colon cancer. The same results were obtained in 11 of 13 patients (84.6%) between immunohistochemical study and RT-PCR-SSCP study. Abnormality of p53 would be a good tumor marker because aberrant expression of p53 protein was found at a rate of more than 60% in each organ. The RT-PCR-SSCP method will be easy to use and reliable in screening for p53 point mutations.

Key words
p53 mutation of colon cancer and gastric cancer, PAb1801, polymerase chain reaction-single stand conformation polymorphism

Reprint requests
Masafumi Kataoka First Department of Surgery, Okayama University Medical School
2-5-1 Shikata-cho, Okayama, 700 JAPAN

Accepted
September 8, 1993

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