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Originally published In Press as doi:10.1074/jbc.M007646200 on January 18, 2001

J. Biol. Chem., Vol. 276, Issue 16, 13153-13159, April 20, 2001
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Interferon gamma  (IFNgamma ) and Tumor Necrosis Factor alpha  Synergism in ME-180 Cervical Cancer Cell Apoptosis and Necrosis
IFNgamma INHIBITS CYTOPROTECTIVE NF-kappa B THROUGH STAT1/IRF-1 PATHWAYS*

Kyoungho SukDagger §||, Inik ChangDagger ||, Yun-Hee Kim**, Sunshin Kim**, Ja Young Kim**, Hocheol Kim§, and Myung-Shik LeeDagger **Dagger Dagger

From the Dagger  Clinical Research Center, Samsung Biomedical Research Institute and ** Department of Medicine, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul 135-710, Korea and § Graduate School of East-West Medical Science, Kyunghee University, Seoul, 130-701, Korea

We investigated the molecular mechanism of the synergism between interferon gamma  (IFNgamma ) and tumor necrosis factor alpha  (TNFalpha ) documented in a variety of biological occasions such as tumor cell death and inflammatory responses. IFNgamma /TNFalpha synergistically induced apoptosis of ME-180 cervical cancer cells. IFNgamma induced STAT1 phosphorylation and interferon regulatory factor 1 (IRF-1) expression. Transfection of phosphorylation-defective STAT1 inhibited IFNgamma /TNFalpha -induced apoptosis, whereas IRF-1 transfection induced susceptibility to TNFalpha . Dominant-negative Ikappa Balpha transfection sensitized ME-180 cells to TNFalpha . IFNgamma pretreatment attenuated TNFalpha - or p65-induced NF-kappa B reporter activity, whereas it did not inhibit p65 translocation or DNA binding of NF-kappa B. IRF-1 transfection alone inhibited TNFalpha -induced NF-kappa B activity, which was reversed by coactivator p300 overexpression. Caspases were activated by IFNgamma /TNFalpha combination; however, caspase inhibition did not abrogate IFNgamma /TNFalpha -induced cell death. Instead, caspase inhibitors directed IFNgamma /TNFalpha -treated ME-180 cells to undergo necrosis, as demonstrated by Hoechst 33258/propidium iodide staining and electron microscopy. Taken together, our results indicate that IFNgamma and TNFalpha synergistically act to destroy ME-180 tumor cells by either apoptosis or necrosis, depending on caspase activation, and STAT1/IRF-1 pathways initiated by IFNgamma play a critical role in IFNgamma /TNFalpha synergism by inhibiting cytoprotective NF-kappa B. IFNgamma /TNFalpha synergism appears to activate cell death machinery independently of caspase activation, and caspase activation seems to merely determine the mode of cell death.

* This work was supported by National Research Laboratory Grants 2000-N-NL-01-C-232 from the Korea Institute of Science and Technology Evaluation and Planning and by Science Research Center Grants from Korea Science and Engineering Foundation.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Supported by Brain Korea 21 project from the Ministry of Education, Korea.

|| These authors contributed equally to this work.

Dagger Dagger Recipient of Juvenile Diabetes Foundation International Research Grant 1-1999-760). To whom correspondence should be addressed. Tel.: 82-2-3410-3436; Fax: 82-2-3410-3849; E-mail: mslee@smc.samsung.co.kr.

Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
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