p53 Detection in Breast Cancer

  • Penelope L. Davis
  • J. Dirk lglehart
Part of the Methods in Molecular Medicine™ book series (MIMM, volume 14)

Abstract

p53 1s a nuclear phosphoprotem whose function is classified as a tumor suppressor (1) Mutattons in the p53 gene are currently regarded as the most common genetic alteration in human cancer (2), including breast cancer (reviewed in ref. 3). Most of these are point mutations within highly conserved regions of the gene that produce altered protein wtth increased stability (4), allowing easy detection in affected cells by in immunohistochemical and immnunoblotting techniques. The presence of elevated levels of mutant p53 may itself be a prognostic factor in human breast cancer (5,6). Furthermore, a significant associatton between high levels of p53 and established pathologrcal crtteria (e.g, tumor stage, estrogen, and progesterone receptor levels) have been described in a number of studies (5,6). Thus chapter will describe two procedures routinely used in our laboratory for the detection of p53 protein in mammary eptthehal cell lines and in breast-tumor tissue. The methods have been used to analyze p53 levels in breast tumors (5, 6, 7) and used in many studies to monitor p53 expression following DNA damage in cell lines (8, 9, 10).

Keywords

Dust Microwave Estrogen Citrate Fluoride 

References

  1. 1.
    Levine A J, Momand J, and Fmlay C. A (1991) The p53 tumour supressor gene Nature 351, 453–456PubMedCrossRefGoogle Scholar
  2. 2.
    Holistem M., Sidransky D, Vogeistem B, and Hans C C. (1991) P53 mutationsinhuman cancer Science 235, 49–53.Google Scholar
  3. 3.
    Harris A L (1992) P53 expressioninhuman breast cancer Adv Cancer Res 59, 69–88PubMedCrossRefGoogle Scholar
  4. 4.
    Caron de Fromentel C and SOUSSI T. (1992) P53 tumour supressor gene a model for investtgating human mutagenesis Genes Chrom Cancer 4, 1–15CrossRefGoogle Scholar
  5. 5.
    Marks J M., Humphrey P. A, Wu K., Berry D., Bandarenko N., Kerns B. M, and Iglehart J. D (1994) Overexpresston of p53 and HER-2neu proteins as prognostic markersinearly stage breast cancer Ann Surg 219, 332–341PubMedCrossRefGoogle Scholar
  6. 6.
    Davidoff A M., Herndon J E, Glover N S, Kerns B. M, Pence J C, Iglehart J D., and Marks J R (1991) Relation between p53 overexpresston and established prognostic factors in breast cancer. Surgery 110, 259–264PubMedGoogle Scholar
  7. 7.
    Bayer E A., Skuteisky E, and Wtlchek M. (1979) The avtdm-btotmin complex in affinity cytochemistry Meth Enzymol. 62, 308–315.PubMedCrossRefGoogle Scholar
  8. 8.
    Elbendary A, Berchuck A, Davis P. L., Havnlesky L, Bast J. C, Iglehart J. D, and Marks J. R. (1994) Transforming growth factor b1 can induce CIPl/WAFl expression independent of the p53 pathway in ovarian cancer celis. Cell Growth Differ 5, 1301–1307PubMedGoogle Scholar
  9. 9.
    Gudas J., Nguyen H, Li T, Hill D., and Cowan K. H. (1995) Effects of cell cycle, wtldtype p53 and DNA damage on p21 expression in human breast epithe ha1 celis Oncogene 11, 253–261PubMedGoogle Scholar
  10. 10.
    Kastan M. B., Onyekwere O, Srdransky D, Voglestein B., and Craig R. (1991) Particrpatton of p53 proteininthe cellular response to DNA damage. Cancer Res 51, 6304–6311PubMedGoogle Scholar
  11. 11.
    Kerns B. M, Jordan P A, Moore M. H, Humphrey P A., Berchuck A, Kohler M F, Bast R C, Iglehart J D., and Marks J R. (1992) P53 over-expressioninformalm-fixed, paraffin-embedded tissue detected by immunohts-tochemistry. J Hutochem Cytochem. 40, 1047–1051.CrossRefGoogle Scholar
  12. 12.
    Towbm H, Staehlm T, and Gordon J (1979) Electrophorettc transfer of proteins from polyacrylamide geis to nitrocellulose sheets procedure and some applications Proc Natl Acad Sci USA 76, 435–4354CrossRefGoogle Scholar
  13. 13.
    Whttehead T. P. (1979) Analyttcal luminescence its potenttalinthe clinical laboratory Clan Chem. 25, 153l–1546Google Scholar
  14. 14.
    Barret A. J and Salvesen G, eds. (1986) Protemase mhibttors,in Research Monographs in Cell and Tzssue Physzology, vol 12 Eisevler, Amsterdam, The Netherlands, pp. 3–18Google Scholar

Copyright information

© Humana Press Inc, Totowa, NJ 1998

Authors and Affiliations

  • Penelope L. Davis
    • 1
  • J. Dirk lglehart
    • 1
  1. 1.Duke University Medical CenterDurham

Personalised recommendations