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Originally published In Press as doi:10.1074/jbc.M313141200 on December 29, 2003

J. Biol. Chem., Vol. 279, Issue 11, 10822-10828, March 12, 2004
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Tumor Necrosis Factor-induced Nonapoptotic Cell Death Requires Receptor-interacting Protein-mediated Cellular Reactive Oxygen Species Accumulation*

Yong Lin{ddagger}§, Swati Choksi{ddagger}, Han-Ming Shen{ddagger}, Qing-Feng Yang{ddagger}, Gang Min Hur{ddagger}, You Sun Kim{ddagger}, Jamie Hong Tran{ddagger}, Sergei A. Nedospasov¶, and Zheng-gang Liu{ddagger}

From the {ddagger}Cell and Cancer Biology Branch, Center for Cancer Research, NCI, National Institutes of Health, Bethesda, Maryland 20892 and Basic Research Program, Science Applications International Corporation Frederick, LMI, CCR, NCI, National Institutes of Health, Frederick, Maryland 21702

The mechanism of tumor necrosis factor (TNF)-induced nonapoptotic cell death is largely unknown, although the mechanism of TNF-induced apoptosis has been studied extensively. In wild-type mouse embryonic fibroblast cells under a caspase-inhibited condition, TNF effectively induced cell death that morphologically resembled necrosis. In this study, we utilized gene knockout mouse embryonic fibroblasts cells and found that tumor necrosis factor receptor (TNFR) I mediates TNF-induced necrotic cell death, and that RIP, FADD, and TRAF2 are critical components of the signaling cascade of this TNF-induced necrotic cell death. Inhibitors of NF-{kappa}B facilitated TNF-induced necrotic cell death, suggesting that NF-{kappa}B suppresses the necrotic cell death pathway. JNK, p38, and ERK activation seem not to be required for this type of cell death because mitogen-activated protein kinase inhibitors did not significantly affect TNF-induced necrotic cell death. In agreement with the previous reports that the reactive oxygen species (ROS) may play an important role in this type of cell death, the ROS scavenger butylated hydroxyanisole efficiently blocked TNF-induced necrotic cell death. Interestingly, during TNF-induced necrotic cell death, the cellular ROS level was significantly elevated in wild type, but not in RIP-/-, TRAF2-/-, and FADD-/- cells. These results suggest that RIP, TRAF2, and FADD are crucial in mediating ROS accumulation in TNF-induced necrotic cell death.

Received for publication, December 2, 2003 , and in revised form, December 22, 2003.

* 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: Cell and Cancer Biology Branch, NCI, National Institutes of Health, Bldg. 10, Rm. 6N105, 9000 Rockville Pike, Bethesda, MD 20892. Tel.: 301-496-3390; Fax: 301-402-1997; E-mail: linyo{at}mail.nih.gov.


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