title: Identification and quantification of concentration-dependent biomarkers in MCF-7/BOS cells exposed to 17β-estradiol by 2-D DIGE and label-free proteomics.
abstract: This paper reports the identification of biomarkers resulting from the exposure of MCF-7/BOS cells to 17β-estradiol (E(2)). The biomarkers were identified using 2 independent and complementary techniques, 2-D DIGE/MALDI-TOF peptide mass fingerprint, and 2-D UPLC-ESI MS/MS. They were identified from the cytosolic fractions of cells treated for 24h with mitogenic concentrations of 1, 30 and 500 pM of 17β-estradiol. Five biomarkers were up-regulated proteins, namely HSP 74, EF2, FKBP4, EF1 and GDIB and one was a down-regulated protein, namely K2C8. Three of these proteins, EF2, FKBP4 and K2C8 are implicated in a network centered on the estrogen receptors ESR1 and ESR2 as well as on AKT1. After the discovery phase, three biomarkers were selected to test the presence of estrogens using selected reaction monitoring (SRM). They were monitored using SRM after incubation of MCF-7/BOS in the presence of E(2) for confirmation or selected xenoestrogens. Daidzein, coumestrol and enterolactone induced an up-regulation of EF2 and FKPB4 proteins, while tamoxifen and resveratrol induced a down-regulation. The exposure of all phytoestrogens induced the down-regulation of K2C8. These markers form a preliminary molecular signature that can be used when testing the estrogenic activity of xenobiotics, either pure or in mixtures.
authors.affiliation: Laboratory of Histology-Cytology, GIGA-R and CART, University of Liège, Allée de la Chimie 3, Sart-Tilman, B 4000 Liège, Belgium.
authors.forename: Mike
authors.initials: M
authors.lastname: Collodoro
authors.forename: Pascale
authors.initials: P
authors.lastname: Lemaire
authors.forename: Gauthier
authors.initials: G
authors.lastname: Eppe
authors.forename: Virginie
authors.initials: V
authors.lastname: Bertrand
authors.forename: Rowan
authors.initials: R
authors.lastname: Dobson
authors.forename: Gabriel
authors.initials: G
authors.lastname: Mazzucchelli
authors.forename: Joëlle
authors.initials: J
authors.lastname: Widart
authors.forename: Edwin
authors.initials: E
authors.lastname: De Pauw
authors.forename: Marie-Claire
authors.initials: MC
authors.lastname: De Pauw-Gillet
mesh_terms: D015415:Biomarkers / Q000032:analysis; D045744:Cell Line, Tumor; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D004958:Estradiol / Q000494:pharmacology*; D006801:Humans; D009374:Neoplasms, Experimental / Q000378:metabolism*; D010449:Peptide Mapping / Q000379:methods*; D020543:Proteome / Q000032:analysis*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D013194:Staining and Labeling
chemical_list: D015415:Biomarkers; D020543:Proteome; D004958:Estradiol