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J. Biol. Chem., Vol. 278, Issue 31, 29184-29191, August 1, 2003

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Necrotic Cell Death in Response to Oxidant Stress Involves the Activation of the Apoptogenic Caspase-8/Bid Pathway^*

Xue Wang , Stefan W. Ryter , Chunsun Dai , Zi-Lue Tang , Simon C. Watkins ¶, Xiao-Ming Yin , Ruiping Song  and Augustine M. K. Choi  ||

From the Division of Pulmonary, Allergy, and Critical Care Medicine, Department of Medicine, the Department of Pathology, and the ^¶Center for Biologic Imaging, Department of Cell Biology and Physiology, School of Medicine, University of Pittsburgh, Pittsburgh, Pennsylvania 15213

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.

Received for publication, February 14, 2003 , and in revised form, May 14, 2003.

^* This work was supported by National Institutes of Health Grants R01-HL60234, R01-AI42365, and R01-HL55330 (to A. M. K. C.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

^|| To whom correspondence should be addressed: Dept. of Medicine, Division of Pulmonary, Allergy, and Critical Care Medicine, 3459 Fifth Ave., Montefiore University Hospital, 628 NW, Pittsburgh, PA 15213. Tel.: 412-692-2210; Fax: 412-692-2260; E-mail: Choiam{at}msx.upmc.edu.

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