Abstract
Human epithelial (A549) cells exposed to hyperoxia die by cellular necrosis. In the current study, we demonstrated the involvement of apoptogenic factors in epithelial cell necrosis in response to hyperoxia, including the formation of the Fas-related death-inducing signaling complex and initiation of mitochondria-dependent apoptotic pathways. We showed increased activation of both Bid and Bax in A549 cells subjected to hyperoxia. Bax activation involved a Bid-assisted conformational change. We discovered that the response to hyperoxia in vivo predominantly involved the activation of the Bid/caspase-8 pathway without apparent increases in Bax expression. Disruption of the Bid pathway by gene deletion protected against cell death in vivo and in vitro. Likewise, inhibition of caspase-8 by Flip also protected against cell death. Taken together, we have demonstrated the involvement of apoptogenic factors in epithelial cell responses to hyperoxia, despite a final outcome of cellular necrosis. We have, for the first time, identified a predominant role for the caspase-8/Bid pathway in signaling associated with hyperoxic lung injury and cell death in vivo and in vitro.
Publication types
-
Research Support, U.S. Gov't, P.H.S.
MeSH terms
-
Adenocarcinoma
-
Animals
-
Apoptosis*
-
BH3 Interacting Domain Death Agonist Protein
-
CASP8 and FADD-Like Apoptosis Regulating Protein
-
Carrier Proteins / genetics
-
Carrier Proteins / metabolism*
-
Carrier Proteins / physiology
-
Caspase 8
-
Caspase 9
-
Caspase Inhibitors
-
Caspases / metabolism*
-
Cells, Cultured
-
Enzyme Inhibitors / pharmacology
-
Fas Ligand Protein
-
Fibroblasts
-
Gene Deletion
-
Humans
-
Intracellular Signaling Peptides and Proteins*
-
Lung
-
Lung Neoplasms
-
Male
-
Membrane Glycoproteins / deficiency
-
Membrane Glycoproteins / physiology
-
Mice
-
Mice, Inbred C3H
-
Mice, Inbred C57BL
-
Mice, Knockout
-
Oxidative Stress*
-
Oxygen / administration & dosage
-
Tumor Cells, Cultured
-
fas Receptor / genetics
-
fas Receptor / physiology
Substances
-
BH3 Interacting Domain Death Agonist Protein
-
BID protein, human
-
Bid protein, mouse
-
CASP8 and FADD-Like Apoptosis Regulating Protein
-
CFLAR protein, human
-
Carrier Proteins
-
Caspase Inhibitors
-
Cflar protein, mouse
-
Enzyme Inhibitors
-
FASLG protein, human
-
Fas Ligand Protein
-
Fasl protein, mouse
-
Intracellular Signaling Peptides and Proteins
-
Membrane Glycoproteins
-
fas Receptor
-
CASP8 protein, human
-
CASP9 protein, human
-
Casp8 protein, mouse
-
Casp9 protein, mouse
-
Caspase 8
-
Caspase 9
-
Caspases
-
Oxygen