Skip to main content

Advertisement

Log in

Discovery and verification of protein differences between Er positive/Her2/neu negative breast tumor tissue and matched adjacent normal breast tissue

  • Preclinical study
  • Published:
Breast Cancer Research and Treatment Aims and scope Submit manuscript

Abstract

This study was designed to quantify and identify differences in protein levels between tumor and adjacent normal breast tissue from the same breast in 18 women with stage I/II ER positive/Her2/neu negative invasive breast cancer. Eighteen separate difference gel electrophoresis (DIGE) gels were run (1 gel per patient). Relative quantification was based on DIGE analysis. After excision and tryptic digestion, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and peptide mass mapping were used to identify protein spots. Two hundred and forty-three spots were differentially abundant between normal and cancer tissues. Fifty spots were identified: 41 were over abundant and nine were less abundant in cancers than in normal breast tissue. Western blotting provided independent confirmation for three of the most biologically and statistically interesting proteins. All 18 gels were replicated by another technician and 32% of the differentially abundant proteins were verified by the duplicate analysis. Follow-up studies are now examining these proteins as biomarkers in blood.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  1. Cancer Facts and Figures – 2007 (2007) American Cancer Society (ACS), Atlanta. www.seer.cancer.gov

  2. Wessel D, Flugge UI (1984) A method for the quantitative recovery of protein in dilute solution in the presence of detergents and lipids. Anal Biochem 138(1):141–143

    Article  CAS  PubMed  Google Scholar 

  3. Bradford MM (1976) Rapid and sensitive method for quantitation of microgram quantities of protein utilizing principle of protein-dye binding. Anal Biochem 72:248–254

    Article  CAS  PubMed  Google Scholar 

  4. Brown LM, Helmke SM, Hunsucker SW et al (2006) Quantitative and qualitative differences in protein expression between papillary thyroid carcinoma and normal thyroid tissue. Mol Carcinog 45(8):613–626

    Article  CAS  PubMed  Google Scholar 

  5. Decyder 2D Software User Manual 28-4010—06 AA. GE Healthcare Amersham Biosciences AB. Bjorkgatan 30. 751 84 Uppsala, Sweden

  6. Rosenfeld J, Capdevielle J, Guillemot JC et al (1992) In-gel digestion of proteins for internal sequence analysis after one- or two-dimensional gel electrophoresis. Anal Biochem 203(1):173–179

    Article  CAS  PubMed  Google Scholar 

  7. Henzel WJ, Billeci TM, Stults JT et al (1993) Identifying proteins from two-dimensional gels by molecular mass searching of peptide fragments in protein sequence databases. Proc Natl Acad Sci USA 90:5011–5015

    Article  CAS  PubMed  Google Scholar 

  8. James P, Quadroni M, Carafoli E et al (1993) Protein identification by mass profile fingerprinting. Biochem Biophys Res Commun 195:58–64

    Article  CAS  PubMed  Google Scholar 

  9. Legrand R, Falconnet JB, Prevost D et al (1993) Peptide mapping of recombinant human interferon-gamma by reversed-phase liquid chromatography with on-line identification by thermospray mass spectrometry and UV absorption spectrometry. J Chromatogr 647(1):3–12

    Article  CAS  PubMed  Google Scholar 

  10. Pappin DJ, Hojrup P, Bleasby AJ (1993) Rapid identification of proteins by peptide-mass fingerprinting. Curr Biol 3(6):327–332

    Article  CAS  PubMed  Google Scholar 

  11. Yates JR III, Speicher S, Griffin PR, Hunkapiller T (1993) Peptide mass maps: a highly informative approach to protein identification. Anal Biochem 214(2):397–408

    Article  CAS  PubMed  Google Scholar 

  12. Netea-Maier RT, Hunsucker SW, Hoevenaars BM et al (2008) Discovery and validation of protein abundance differences between follicular thyroid neoplasms. Cancer Res 68(5):1572–1580

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Lindsay-Rae B. Weitzel.

Additional information

M. W. Duncan and A. Elias contributed equally to this work.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (DOC 50 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Weitzel, LR.B., Byers, T., Allen, J. et al. Discovery and verification of protein differences between Er positive/Her2/neu negative breast tumor tissue and matched adjacent normal breast tissue. Breast Cancer Res Treat 124, 297–305 (2010). https://doi.org/10.1007/s10549-009-0721-6

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10549-009-0721-6

Keywords

Navigation