Identification and quantification of concentration-dependent biomarkers in MCF-7/BOS cells exposed to 17¦Â-estradiol by 2-D DIGE and label-free proteomics

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• 作者：Mike Collodoro ; Pascale Lemaire ; Gauthier Eppe ; Virginie Bertr ; Rowan Dobson ; Gabriel Mazzucchelli ; Jo?lle Widart ; Edwin De Pauw ; Marie-Claire De Pauw-Gillet
• 关键词：17¦Â-Estradiol ; MCF-7 BOS ; 2-D DIGE ; Label-free ; SRM ; Xenoestrogens
• 刊名：Journal of Proteomics
• 出版年：2012
• 出版时间：19 July, 2012
• 年：2012
• 卷：75
• 期：14
• 页码：4555-4569
• 全文大小：1541 K

文摘

This paper reports the identification of biomarkers resulting from the exposure of MCF-7/BOS cells to 17¦Â-estradiol (E₂). 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 24 h with mitogenic concentrations of 1, 30 and 500 pM of 17¦Â-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 E₂ 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. This article is part of a Special Issue entitled: Farm animal proteomics.