title: Quantitative proteomic determination of diethylstilbestrol action on prostate cancer.
abstract: 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).
authors: Department of Urology, Angers University Hospital, Angers 49933, France. pibigot@chu-angers.fr
authors: Pierre
authors: P
authors: Bigot
authors: Kevin
authors: K
authors: Mouzat
authors: Souhil
authors: S
authors: Lebdai
authors: Muriel
authors: M
authors: Bahut
authors: Nora
authors: N
authors: Benhabiles
authors: Géraldine Cancel
authors: GC
authors: Tassin
authors: Abdel-Rahmène
authors: AR
authors: Azzouzi
authors: Olivier
authors: O
authors: Cussenot
mesh_terms: D045744:Cell Line, Tumor; D004054:Diethylstilbestrol / Q000494:pharmacology*; D004968:Estrogens, Non-Steroidal / Q000494:pharmacology*; D015870:Gene Expression / Q000187:drug effects*; D055551:HSP27 Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism; D006360:Heat-Shock Proteins; D006801:Humans; D008297:Male; D018832:Molecular Chaperones; D009953:Ornithine-Oxo-Acid Transaminase / Q000235:genetics / Q000378:metabolism; D011471:Prostatic Neoplasms / Q000235:genetics / Q000378:metabolism*; D020543:Proteome / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D040901:Proteomics
chemical_list: D004968:Estrogens, Non-Steroidal; D055551:HSP27 Heat-Shock Proteins; C057725:HSPB1 protein, human; D006360:Heat-Shock Proteins; D018832:Molecular Chaperones; D020543:Proteome; D004054:Diethylstilbestrol; D009953:Ornithine-Oxo-Acid Transaminase