Quantitative proteomic determination of diethylstilbestrol action on prostate cancer

作     者：Pierre Bigot Kevin Mouzat Muriel Bahut Nora Benhabiles Geraldine Cancel Tassin Abdel-Rahmene Azzouzi Olivier Cussenot 

作者机构：Department of Urology Angers University Hospital Angers 49933 France CeRePP Tenon Hospital Paris 75970 France Department of Biochemistry N'imes University Hospital NTmes 30029 France Plate-Forme Technologique de Biotechnologie Moleculaire Angers University Angers 49933 France 5CEA DSV IRCM SREIT Laboratoire de Cancerologie Experimentale Fontenay-aux-Roses 92265 France 

出 版 物：《Asian Journal of Andrology》 (亚洲男性学杂志（英文版）)

年 卷 期：2013年第15卷第3期

页      面：413-420页

核心收录：

学科分类：0710[理学-生物学] 071010[理学-生物化学与分子生物学] 081704[工学-应用化学] 07[理学] 08[工学] 0817[工学-化学工程与技术] 09[农学] 071007[理学-遗传学] 0901[农学-作物学] 0836[工学-生物工程] 090102[农学-作物遗传育种] 

基　　金：the Association Francaise d'Urologie (AFU) the Association pour la recherche sur les tumeurs de la prostate (ARTP) 

主　　题：cultured cells DES diethylstilbestrol ingenuity pathway analysis isotope labelling mass spectrometry prostate cancer proteomics 

摘      要：Diethylstilbestrol （DES） has a direct cellular mechanism inhibition on prostate cancer. Its action is independent from the oestrogen receptors and is preserved after a first-line hormonal therapy. We aimed to identify proteins involved in the direct cellular inhibition effects of DES on prostate cancer. We used a clonogenic assay to establish the median lethal concentration of DES on 22RV1 cells. 22RV1 cells were exposed to standard and DES-enriched medium. After extraction, protein expression levels were obtained by two-dimensional differential in-gel electrophoresis （2D-DIGE） and isotope labelling tags for relative and absolute quantification （iTRAQ）. Proteins of interest were analysed by quantitative RT-PCR and western blotting. The differentially regulated proteins （P〈0.01） were interrogated against a global molecular network based on the ingenuity knowledge base. The 2D-DIGE analyses revealed DES-induced expression changes for 14 proteins （〉 1.3 fold; P〈0.05）. The iTRAQ analyses allowed the identification of 895 proteins. Among these proteins, 65 had a modified expression due to DES exposure （i.e., 23 overexpressed and 42 underexpressed）. Most of these proteins were implicated in apoptosis and redox processes and had a predicted mitochondrial expression. Additionally, ingenuity pathway analysis placed the OAT and HSBP1 genes at the centre of a highly significant network. RT-PCR confirmed the overexpression of OAT （P=0.006） and HSPB1 （P=0.046）.