title: Discovery and verification of protein differences between Er positive/Her2/neu negative breast tumor tissue and matched adjacent normal breast tissue.
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.
authors.affiliation: Department of Preventive Medicine, University of Colorado Denver, Anschutz Medical Campus, Aurora, CO, USA. Lindsay.Weitzel@ucdenver.edu
authors.forename: Lindsay-Rae B
authors.initials: LR
authors.lastname: Weitzel
authors.forename: Tim
authors.initials: T
authors.lastname: Byers
authors.forename: Jenna
authors.initials: J
authors.lastname: Allen
authors.forename: Christina
authors.initials: C
authors.lastname: Finlayson
authors.forename: Steve M
authors.initials: SM
authors.lastname: Helmke
authors.forename: John E
authors.initials: JE
authors.lastname: Hokanson
authors.forename: Stephen W
authors.initials: SW
authors.lastname: Hunsucker
authors.forename: James R
authors.initials: JR
authors.lastname: Murphy
authors.forename: Keri
authors.initials: K
authors.lastname: Newell
authors.forename: Kelly M
authors.initials: KM
authors.lastname: Queensland
authors.forename: Meenakshi
authors.initials: M
authors.lastname: Singh
authors.forename: Paul E
authors.initials: PE
authors.lastname: Wischmeyer
authors.forename: Mark W
authors.initials: MW
authors.lastname: Duncan
authors.forename: Anthony
authors.initials: A
authors.lastname: Elias
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D014408:Biomarkers, Tumor / Q000032:analysis*; D015153:Blotting, Western; D001943:Breast Neoplasms / Q000737:chemistry* / Q000473:pathology; D030562:Databases, Protein; D015180:Electrophoresis, Gel, Two-Dimensional; D005260:Female; D006801:Humans; D008875:Middle Aged; D009361:Neoplasm Invasiveness; D009367:Neoplasm Staging; D015588:Observer Variation; D010449:Peptide Mapping; D040901:Proteomics / Q000379:methods; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000032:analysis*; D011960:Receptors, Estrogen / Q000032:analysis*; D015203:Reproducibility of Results; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D014408:Biomarkers, Tumor; D018719:Erb-b2 Receptor Tyrosine Kinases; D011960:Receptors, Estrogen; C508053:ERBB2 protein, human