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Related Experiment Videos

p53 protein immunostaining in routinely processed paraffin-embedded sections

J M Bruner1, J H Connelly, H Saya

  • 1Department of Pathology, University of Texas M. D. Anderson Cancer Center, Houston.

Modern Pathology : an Official Journal of the United States and Canadian Academy of Pathology, Inc
|March 1, 1993
PubMed
Summary

Detecting mutant p53 protein in archived tumors is now possible using a new paraffin-embedded section immunostaining technique. This method shows high concordance with frozen sections, offering a valuable screening tool for cancer research.

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Area of Science:

  • Oncology
  • Molecular Biology
  • Pathology

Background:

  • Mutation of the p53 tumor suppressor gene is a frequent event in human cancers.
  • Altered p53 protein accumulates and can be detected via immunohistochemistry.
  • Standard detection methods often require fresh tissue, limiting the use of archived samples.

Purpose of the Study:

  • To develop and validate a novel immunostaining technique for detecting mutant p53 protein in routinely processed paraffin-embedded human tumor tissues.
  • To compare the concordance of this new method with traditional frozen-section immunostaining.
  • To assess the utility of this technique for screening p53 mutations in archival tumor specimens.

Main Methods:

  • Immunohistochemistry was performed on paraffin-embedded tissue sections from human gliomas and colorectal carcinomas.

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  • A monoclonal antibody against the altered p53 protein was used for staining.
  • Results were compared with immunostaining on corresponding frozen sections.
  • Tumor cell lines with known p53 alterations served as controls.

    • Main Results:

    • The paraffin-embedded section method demonstrated high concordance with frozen sections: 100% in colorectal carcinomas and 83% in gliomas.
    • No false positives were observed in paraffin-embedded sections.
    • A small discrepancy was noted in some gliomas, potentially due to technical factors like fixation time.

    Conclusions:

    • A reliable paraffin-embedded section immunostaining technique for detecting mutant p53 protein has been established.
    • This method is suitable for screening p53 mutations in archived human tumors.
    • The technique holds potential for correlation with clinical data, aiding in cancer diagnosis and outcome prediction.