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Immunohistochemical detection of p53 overexpression in formalin-fixed, paraffin-embedded sections of endometrial
K Iwai1, T Hachisuga, T Iwasaka
1Department of Obstetrics and Gynecology, Saga Medical School, Japan.
Objective:
In the past, immunohistochemical detection of p53 has been possible only in the case of fresh-frozen tissues. However using CM1 and an enhanced method based on microwave heating and protease digestion enabled the immunohistochemical detection of p53 overexpression in formalin-fixed, paraffin-embedded sections of endometrial carcinoma, although very few positive stainings were obtained without such an enhanced treatment. To evaluate the enhancement method, a comparative study was performed on frozen materials and paraffin-embedded samples.
Methods:
Fifty-three frozen materials, including normal endometrial glands, endometrial hyperplasias and carcinomas, and paraffin-embedded samples from the same patients were used in this study. PAb1801 (mouse monoclonal antibody) was used to analyse p53 overexpression in the frozen sections, while CM1 (rabbit polyclonal anti-p53 antibody) was used for the paraffin sections.
Results:
PAb1801 and CM1 staining on normal endometriums and endometrial lesions showed a significant degree of concordance between frozen and paraffin-embedded materials.
Conclusion:
Enhanced immunodetection of p53 in paraffin-embedded tissues will provide a useful alternative to the usual fresh-tissue assay.
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