SELDI protein profiling of dunning R-3327 derived cell lines: identification of molecular markers of prostate cancer progression

Prostate. 2007 Oct 1;67(14):1565-75. doi: 10.1002/pros.20646.


Background: We recently demonstrated the protein expression profiling of Dunning rat tumor cell lines of varying metastatic potential (G (0%), AT-1 ( approximately 20%), and MLL (100%)) using SELDI-TOF-MS. As a parallel effort, we have been pursuing the identification of the protein(s) comprising the individual discriminatory "peaks" and evaluating their utility as potential biomarkers for prostate cancer progression.

Methods: To identify the observed SELDI-TOF-MS m/z (mass/charge) values with discriminatory expression between different sublines, we employed a combination of chemical pre-fractionation, liquid chromatography, gel electrophoresis and tandem mass spectroscopy. Identified proteins were then verified by immuno-assay and Western analysis.

Results: A 17.5 K m/z SELDI-TOF-MS peak was found to retain discriminatory value in each of two separate study-sets with an increased expression in the metastatic MLL line. Sequence identification and subsequent immunoassays verified that Histone H2B is the observed 17.5 K m/z SELDI peak. SELDI-based immuno-assay and Western Blotting revealed that Histone H2B is specifically over-expressed in metastatic MLL lines.

Conclusions: SELDI-TOF MS analysis of the Dunning prostate cancer cell lines confirmed the consistent overexpression of a 17.5 K m/z peak in metastatic MLL subline. The 17.5 kDa protein from MLL has been isolated and identified as Histone H2B.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Adenocarcinoma / chemistry
  • Adenocarcinoma / metabolism*
  • Adenocarcinoma / secondary
  • Animals
  • Biomarkers, Tumor / analysis
  • Biomarkers, Tumor / metabolism*
  • Cell Line, Tumor
  • Chromatography, High Pressure Liquid
  • Disease Progression
  • Electrophoresis, Polyacrylamide Gel
  • Male
  • Neoplasm Metastasis
  • Neoplasm Proteins / analysis
  • Neoplasm Proteins / metabolism*
  • Prostatic Neoplasms / chemistry
  • Prostatic Neoplasms / metabolism*
  • Prostatic Neoplasms / pathology
  • Protein Array Analysis*
  • Proteomics
  • Rats
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / methods*
  • Tandem Mass Spectrometry


  • Biomarkers, Tumor
  • Neoplasm Proteins