Skip to main content
NCBI home page
British Journal of Cancer logoLink to British Journal of Cancer
. 1993 Jun;67(6):1254–1258. doi: 10.1038/bjc.1993.234

Comparison between p53 staining in tissue sections and p53 proteins levels measured by an ELISA technique.

B Vojtĕsek 1, C J Fisher 1, D M Barnes 1, D P Lane 1
PMCID: PMC1968521  PMID: 7685617

Abstract

We studied 51 paired samples of tissue sections and cytosol extracts from patients with breast cancer. A very high affinity monoclonal antibody to human p53 protein, DO-1, and polyclonal serum CM-1 to p53 protein were used for two site ELISA assays and CM-1 was used for immunohistochemistry to detect p53 protein accumulation in breast cancer samples. Eighteen carcinomas were positive for p53 by tissue staining and ELISA assay. Nineteen tumours were negative by ELISA and immunohistochemistry, and 14 cases with low levels of positive staining by immunohistochemistry were negative by the ELISA assay. A statistically significant correlation has been found between the degree of staining and the amount of p53 protein measured by ELISA (Pearson's correlation coefficient r = 0.59, P < 0.00001). Our ELISA assay offers an alternative approach to evaluating the p53 status of breast biopsy material, using cytosol extracts routinely prepared for steroid hormone receptor assays. This assay should also be of general application to other situations where the level of p53 protein needs to be determined.

Full text

PDF
1254

Images in this article

1255Figure 1
on p.1255

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Ahuja H., Bar-Eli M., Advani S. H., Benchimol S., Cline M. J. Alterations in the p53 gene and the clonal evolution of the blast crisis of chronic myelocytic leukemia. Proc Natl Acad Sci U S A. 1989 Sep;86(17):6783–6787. doi: 10.1073/pnas.86.17.6783. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Baker S. J., Fearon E. R., Nigro J. M., Hamilton S. R., Preisinger A. C., Jessup J. M., vanTuinen P., Ledbetter D. H., Barker D. F., Nakamura Y. Chromosome 17 deletions and p53 gene mutations in colorectal carcinomas. Science. 1989 Apr 14;244(4901):217–221. doi: 10.1126/science.2649981. [DOI] [PubMed] [Google Scholar]
  3. Bartek J., Iggo R., Gannon J., Lane D. P. Genetic and immunochemical analysis of mutant p53 in human breast cancer cell lines. Oncogene. 1990 Jun;5(6):893–899. [PubMed] [Google Scholar]
  4. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  5. Callahan R. p53 mutations, another breast cancer prognostic factor. J Natl Cancer Inst. 1992 Jun 3;84(11):826–827. doi: 10.1093/jnci/84.11.826. [DOI] [PubMed] [Google Scholar]
  6. Cattoretti G., Rilke F., Andreola S., D'Amato L., Delia D. P53 expression in breast cancer. Int J Cancer. 1988 Feb 15;41(2):178–183. doi: 10.1002/ijc.2910410204. [DOI] [PubMed] [Google Scholar]
  7. Chiba I., Takahashi T., Nau M. M., D'Amico D., Curiel D. T., Mitsudomi T., Buchhagen D. L., Carbone D., Piantadosi S., Koga H. Mutations in the p53 gene are frequent in primary, resected non-small cell lung cancer. Lung Cancer Study Group. Oncogene. 1990 Oct;5(10):1603–1610. [PubMed] [Google Scholar]
  8. Davidoff A. M., Herndon J. E., 2nd, Glover N. S., Kerns B. J., Pence J. C., Iglehart J. D., Marks J. R. Relation between p53 overexpression and established prognostic factors in breast cancer. Surgery. 1991 Aug;110(2):259–264. [PubMed] [Google Scholar]
  9. Davidoff A. M., Humphrey P. A., Iglehart J. D., Marks J. R. Genetic basis for p53 overexpression in human breast cancer. Proc Natl Acad Sci U S A. 1991 Jun 1;88(11):5006–5010. doi: 10.1073/pnas.88.11.5006. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Davidoff A. M., Iglehart J. D., Marks J. R. Immune response to p53 is dependent upon p53/HSP70 complexes in breast cancers. Proc Natl Acad Sci U S A. 1992 Apr 15;89(8):3439–3442. doi: 10.1073/pnas.89.8.3439. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hall P. A., Ray A., Lemoine N. R., Midgley C. A., Krausz T., Lane D. P. p53 immunostaining as a marker of malignant disease in diagnostic cytopathology. Lancet. 1991 Aug 24;338(8765):513–513. doi: 10.1016/0140-6736(91)90586-e. [DOI] [PubMed] [Google Scholar]
  12. Hopwood D., Slidders W., Yeaman G. R. Tissue fixation with phenol-formaldehyde for routine histopathology. Histochem J. 1989 Apr;21(4):228–234. doi: 10.1007/BF01747525. [DOI] [PubMed] [Google Scholar]
  13. Iggo R., Gatter K., Bartek J., Lane D., Harris A. L. Increased expression of mutant forms of p53 oncogene in primary lung cancer. Lancet. 1990 Mar 24;335(8691):675–679. doi: 10.1016/0140-6736(90)90801-b. [DOI] [PubMed] [Google Scholar]
  14. Levine A. J., Momand J., Finlay C. A. The p53 tumour suppressor gene. Nature. 1991 Jun 6;351(6326):453–456. doi: 10.1038/351453a0. [DOI] [PubMed] [Google Scholar]
  15. Lu X., Park S. H., Thompson T. C., Lane D. P. Ras-induced hyperplasia occurs with mutation of p53, but activated ras and myc together can induce carcinoma without p53 mutation. Cell. 1992 Jul 10;70(1):153–161. doi: 10.1016/0092-8674(92)90541-j. [DOI] [PubMed] [Google Scholar]
  16. McGuire W. L. Breast cancer prognostic factors: evaluation guidelines. J Natl Cancer Inst. 1991 Feb 6;83(3):154–155. doi: 10.1093/jnci/83.3.154. [DOI] [PubMed] [Google Scholar]
  17. Midgley C. A., Fisher C. J., Bártek J., Vojtesek B., Lane D., Barnes D. M. Analysis of p53 expression in human tumours: an antibody raised against human p53 expressed in Escherichia coli. J Cell Sci. 1992 Jan;101(Pt 1):183–189. doi: 10.1242/jcs.101.1.183. [DOI] [PubMed] [Google Scholar]
  18. Nigro J. M., Baker S. J., Preisinger A. C., Jessup J. M., Hostetter R., Cleary K., Bigner S. H., Davidson N., Baylin S., Devilee P. Mutations in the p53 gene occur in diverse human tumour types. Nature. 1989 Dec 7;342(6250):705–708. doi: 10.1038/342705a0. [DOI] [PubMed] [Google Scholar]
  19. Ostrowski J. L., Sawan A., Henry L., Wright C., Henry J. A., Hennessy C., Lennard T. J., Angus B., Horne C. H. p53 expression in human breast cancer related to survival and prognostic factors: an immunohistochemical study. J Pathol. 1991 May;164(1):75–81. doi: 10.1002/path.1711640113. [DOI] [PubMed] [Google Scholar]
  20. Rodrigues N. R., Rowan A., Smith M. E., Kerr I. B., Bodmer W. F., Gannon J. V., Lane D. P. p53 mutations in colorectal cancer. Proc Natl Acad Sci U S A. 1990 Oct;87(19):7555–7559. doi: 10.1073/pnas.87.19.7555. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Takahashi T., Nau M. M., Chiba I., Birrer M. J., Rosenberg R. K., Vinocour M., Levitt M., Pass H., Gazdar A. F., Minna J. D. p53: a frequent target for genetic abnormalities in lung cancer. Science. 1989 Oct 27;246(4929):491–494. doi: 10.1126/science.2554494. [DOI] [PubMed] [Google Scholar]
  22. Thor A. D., Moore DH I. I., Edgerton S. M., Kawasaki E. S., Reihsaus E., Lynch H. T., Marcus J. N., Schwartz L., Chen L. C., Mayall B. H. Accumulation of p53 tumor suppressor gene protein: an independent marker of prognosis in breast cancers. J Natl Cancer Inst. 1992 Jun 3;84(11):845–855. doi: 10.1093/jnci/84.11.845. [DOI] [PubMed] [Google Scholar]
  23. Vojtesek B., Bártek J., Midgley C. A., Lane D. P. An immunochemical analysis of the human nuclear phosphoprotein p53. New monoclonal antibodies and epitope mapping using recombinant p53. J Immunol Methods. 1992 Jul 6;151(1-2):237–244. doi: 10.1016/0022-1759(92)90122-a. [DOI] [PubMed] [Google Scholar]
  24. Zambetti G. P., Olson D., Labow M., Levine A. J. A mutant p53 protein is required for maintenance of the transformed phenotype in cells transformed with p53 plus ras cDNAs. Proc Natl Acad Sci U S A. 1992 May 1;89(9):3952–3956. doi: 10.1073/pnas.89.9.3952. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from British Journal of Cancer are provided here courtesy of Cancer Research UK

RESOURCES