Mapping cyclosporine-induced changes in protein secretion by renal cells using stable isotope labeling with amino acids in cell culture (SILAC)
| Autor: | Louis Noël Gastinel, Fabien Lamoureux, Marie Essig, Elodie Mestre, Pierre Marquet |
|---|---|
| Přispěvatelé: | Pharmacologie des Immunosuppresseurs et de la Transplantation (PIST), Université de Limoges (UNILIM)-CHU Limoges-Génomique, Environnement, Immunité, Santé, Thérapeutique (GEIST FR CNRS 3503)-Institut National de la Santé et de la Recherche Médicale (INSERM), Service de Pharmacologie, toxicologie et pharmacovigilance [CHU Limoges], CHU Limoges, Service de Néphrologie [Rouen], CHU Rouen, Normandie Université (NU)-Normandie Université (NU)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU), Service de Néphrologie, Dialyse, Transplantations [CHU Limoges], Marquet, Pierre |
| Rok vydání: | 2012 |
| Předmět: |
MESH: Extracellular Matrix Proteins
MESH: Amino Acids MESH: Isotope Labeling Proteome 030232 urology & nephrology Kidney Proteomics MESH: Cyclosporine Biochemistry MESH: Down-Regulation Cyclophilins 0302 clinical medicine Stable isotope labeling by amino acids in cell culture MESH: Up-Regulation Amino Acids Cells Cultured chemistry.chemical_classification Extracellular Matrix Proteins 0303 health sciences MESH: Cyclophilin A [SDV.SP]Life Sciences [q-bio]/Pharmaceutical sciences MESH: Gene Expression Regulation Up-Regulation 3. Good health Amino acid [SDV.SP] Life Sciences [q-bio]/Pharmaceutical sciences MESH: Proteome medicine.anatomical_structure Isotope Labeling MESH: HEK293 Cells Toxicity Cyclosporine MESH: Cell Adhesion Molecules MESH: Immunosuppressive Agents Cyclophilin A Immunosuppressive Agents MESH: Cells Cultured Biophysics Down-Regulation Biology MESH: Cyclophilins Nephrotoxicity 03 medical and health sciences medicine Humans Cell adhesion 030304 developmental biology MESH: Humans MESH: Kidney HEK293 Cells Secretory protein Gene Expression Regulation chemistry Cell Adhesion Molecules |
| Zdroj: | Journal of Proteomics Journal of Proteomics, Elsevier, 2012, 75 (12), pp.3674-87. ⟨10.1016/j.jprot.2012.04.024⟩ |
| ISSN: | 1874-3919 1876-7737 |
| DOI: | 10.1016/j.jprot.2012.04.024 |
| Popis: | International audience; Nephrotoxicity is an adverse event that strongly limits the use of the immunosuppressant cyclosporine in solid organ transplantation and the precise molecular mechanisms underlying this toxicity remain unclear. MS-based proteomic analysis of the secretome of HEK-293 renal cells exposed to cyclosporine was performed to identify changes in protein secretion, as a first step to discover potential biomarkers of such nephrotoxicity. To detect and quantify the perturbed proteins in the culture medium we used SILAC and nano-scale liquid chromatography followed by MALDI-TOF/TOF mass spectrometry. Among 106 proteins identified, 80 were quantified in both forward/reverse SILAC experiments and quantitative proteomic analysis revealed altered levels of expression for 24 secreted proteins. These included the down-regulation of a number of extracellular matrix/cell adhesion components, and the up-regulation of secreted cyclophilins A and B, macrophage inhibition factor and phosphatidylethanolamine-binding protein 1. These changes in protein secretion were not prevented by co-incubation with the antioxidant N-acetylcysteine, suggesting that they were not triggered by cyclosporine-induced oxidative stress. The results from the present study provide important new knowledge to gain insights into the molecular mechanisms of cyclosporine-related toxicity. Some of the proteins identified here should be tested as potential biomarkers of cyclosporine nephrotoxicity in subsequent clinical studies. |
| Databáze: | OpenAIRE |
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