title: Proteomic analysis identifies nuclear protein effectors in PKC-delta signaling under high glucose-induced apoptosis in human umbilical vein endothelial cells.
abstract: Although experimental animal and clinical trials have suggested that additional mechanisms other than protein kinase C (PKC)-β activation are involved in the vascular pathology of diabetic complications, current knowledge on the role of PKC-delta is incomplete and inconclusive. Human umbilical vein endothelial cells (HUVECs) were cultured in both high and normal glucose conditions and infected with recombinant adenovirus to overexpress PKC-delta. PKC-delta expression was also down-regulated using the PKC-delta inhibitor, rottlerin. Using flow cytometric analysis, we showed that PKC-delta is activated and translocates to the nucleus under high glucose conditions. Augmented cell apoptosis and cell cycle arrest were observed in a PKC-delta-dependent manner in the HUVECs. Furthermore, proteomic analyses identified 51 high glucose-induced and PKC-delta-associated proteins, and subsequent matrix-assisted laser desorption/ionization time of flight mass spectrometry analysis uncovered a total of 37 unique proteins. The majority of identified proteins were previously unknown targets of PKC-delta signaling and were involved in the regulation of the cell cycle and apoptosis, tumor suppression, transcription, stress and signal transduction within the nucleus. Our data show that PKC-delta is an important mediator of cell apoptosis and cell cycle arrest in HUVECs under high glucose stress.
authors.affiliation: Department of Endocrinology, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, P.R. China.
authors.forename: Fang
authors.initials: F
authors.lastname: Sun
authors.forename: Bo
authors.initials: B
authors.lastname: Zhou
authors.forename: Xuebo
authors.initials: X
authors.lastname: Lin
authors.forename: Lian
authors.initials: L
authors.lastname: Duan
mesh_terms: D017209:Apoptosis / Q000187:drug effects*; D002453:Cell Cycle / Q000187:drug effects; D002467:Cell Nucleus / Q000187:drug effects / Q000378:metabolism; D015180:Electrophoresis, Gel, Two-Dimensional; D042783:Endothelial Cells / Q000166:cytology* / Q000187:drug effects / Q000201:enzymology; D004789:Enzyme Activation / Q000187:drug effects; D005947:Glucose / Q000494:pharmacology*; D006801:Humans; D009687:Nuclear Proteins / Q000378:metabolism*; D010449:Peptide Mapping; D051745:Protein Kinase C-delta / Q000378:metabolism*; D021381:Protein Transport / Q000187:drug effects; D040901:Proteomics / Q000379:methods*; D015398:Signal Transduction / Q000187:drug effects*; D013312:Stress, Physiological / Q000187:drug effects; D014471:Umbilical Veins / Q000166:cytology; D015854:Up-Regulation / Q000187:drug effects
chemical_list: D009687:Nuclear Proteins; D051745:Protein Kinase C-delta; D005947:Glucose