Chondrocyte cell death mediated by reactive oxygen species-dependent activation of PKC-betaI.
Signals generated by the extracellular matrix (ECM) promote cell survival. We have shown that chondrocytes detached from their native ECM and plated without serum at low density on poly-l-lysine undergo significant cell death that is associated with the production of reactive oxygen species (ROS). No cell death or ROS production was observed when cells were plated on fibronectin under the same conditions. Cell death on poly-l-lysine could be completely inhibited with the addition of either antioxidants or inhibitors of specific protein kinase C (PKC) isoforms including PKC-betaI. PKC-betaI was noted to translocate from the cytosol to the particulate membrane after plating on poly-l-lysine, and this translocation was inhibited by the addition of an antioxidant. Time-course analyses implicated endogenous ROS production as a secondary messenger leading to PKC-betaI activation and subsequent chondrocyte cell death. Cell survival on poly-l-lysine was significantly improved in the presence of oligomycin or DIDS, suggesting that ROS production occurred via complex V of the electron transport chain of the mitochondria and that ROS were released to the cytosol via voltage-dependent anion channels. Together, these results represent a novel mechanism by which ROS can initiate cell death through the activation of PKC-betaI.
DelCarlo
Marcello
M
Department of Medicine, Section of Rheumatology and Department of Biochemistry, Rush Medical College, Chicago, Illinois, USA.
Loeser
Richard F
RF
D000975:Antioxidants / Q000494:pharmacology; D018929:Cell Culture Techniques; D016923:Cell Death; D002478:Cells, Cultured; D019902:Chondrocytes / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D045222:Electron Transport Chain Complex Proteins / Q000037:antagonists & inhibitors; D004789:Enzyme Activation; D005353:Fibronectins; D006801:Humans; D008928:Mitochondria / Q000378:metabolism; D011107:Polylysine; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000378:metabolism*; D064546:Protein Kinase C beta; D051745:Protein Kinase C-delta / Q000037:antagonists & inhibitors / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism*; D013997:Time Factors
D000975:Antioxidants; D045222:Electron Transport Chain Complex Proteins; D005353:Fibronectins; D017382:Reactive Oxygen Species; D011107:Polylysine; D011493:Protein Kinase C; D064546:Protein Kinase C beta; D051745:Protein Kinase C-delta