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Serain, Elena (2008) Uso dell'analisi proteomica differenziale in fase liquida nello studio dell'epressione indotta da selenio e della resistenza al Cisplatino. [Ph.D. thesis]

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The effect of 50 nM, 100 nM sodium selenite and 100 nM selenomethionine supplementation was studied on LNCaP cells by a proteomic approach, on ProteomeLabTM PF 2D platform. Proteins were separated by liquid phase bi-dimensional chromatography and analyzed by pair-wise alignment of peaks to detect those differentially expressed. Differential expression threshold was set at a twice difference level and proteins matching this criterion were identified by MALDI-TOF and confirmed by ESI-Ion Trap MS/MS. Not all differentially expressed proteins found by PF 2DTM could be identified by MS analysis, the sensitivity of which emerging as the limiting factor. Thus, only the most abundant proteins, differently expressed following selenium supplementation, were identified. We positively showed an increase of expression of thioredoxin reductase 1, enolase 1, phosphoglycerate mutase 1, glyceraldehyde-3-phosphate dehydrogenase, heterogeneous nuclear ribonucleoprotein A2/B1, isoform A2, Ras-GTPase-activating protein SH3-domain-binding protein and keratin 18 and a decrease of expression of peroxiredoxin 1 and heat shock 70 kDa protein 8, isoform 1. Results are consistent, at least in part, with the less oxidant environment brought about by the synthesis of Se-dependent peroxidases, keeping low the steady-state concentration of hydrogen peroxide.
The same proteomic approach was applied for studying proteins differentially expressed on 2008 and C13* cells, cisplatin-sensitive and resistant ovarian cancer cells, respectively. Proteins identified by MALDI-TOF analysis were 24 under-expressed on C13* and 20 over-expressed. Glutathione transferase, TSA (peroxiredoxin 2), protein disulfide isomerase, stress-induced-phosphoprotein 1 (Hsp70/Hsp90-organizing protein), thioredoxin domain-containing protein 2 e BCL2-associated athanogene 3, isoform CRA_a were over-expressed, while beta-tubulin and other cytoskeletal proteins were under-expressed. Results are consistent with mechanisms of cisplatin resistance

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EPrint type:Ph.D. thesis
Tutor:Ursini, Fulvio
Ph.D. course:Ciclo 20 > Scuole per il 20simo ciclo > BIOCHIMICA E BIOTECNOLOGIE > BIOCHIMICA E BIOFISICA
Data di deposito della tesi:31 January 2008
Anno di Pubblicazione:31 January 2008
Key Words:Selenium; liquid phase bi-dimensional chromatography; differentially expressed proteins; MALDI-TOF; cisplatin resistance.
Settori scientifico-disciplinari MIUR:Area 05 - Scienze biologiche > BIO/10 Biochimica
Struttura di riferimento:Dipartimenti > pre 2012 Dipartimento di Chimica Biologica
Codice ID:287
Depositato il:22 Oct 2008
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