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*Ovarian Cancer
Endocrinology Vol. 144, No. 2 623-630
Copyright © 2003 by The Endocrine Society

ARTICLE

Resistance of Human Ovarian Cancer Cells to Tumor Necrosis Factor {alpha} Is a Consequence of Nuclear Factor {kappa}B-Mediated Induction of Fas-Associated Death Domain-Like Interleukin-1ß-Converting Enzyme-Like Inhibitory Protein

Chao Wu Xiao, Xiaojuan Yan, Yulian Li, Shrikanth A. G. Reddy and Benjamin K. Tsang

Department of Obstetrics and Gynecology and Cellular and Molecular Medicine (C.W.X., X.Y., Y.L., B.K.T.), Reproductive Biology Unit and Division of Gynecologic Oncology, University of Ottawa, Ottawa Health Research Institute, The Ottawa Hospital, Ottawa, Ontario, Canada K1Y 4E9; Nutrition Research Division (C.W.X.), Food Directorate, Health Products and Food Branch, Health Canada, Banting Research Centre, Ottawa, Ontario, Canada K1A 0L2; Department of GI Oncology and Digestive Diseases (S.A.G.R.), Division of Medicine, M. D. Anderson Cancer Center, University of Texas, Houston, Texas 77030

Address all correspondence and requests for reprints to: Dr. Benjamin K. Tsang, Ottawa Health Research Institute, The Ottawa Hospital (Civic Campus), 725 Parkdale Avenue, Ottawa, Ontario, Canada K1Y 4E9. E-mail: btsang{at}ohri.ca.

The purpose of the present studies was to examine the role and regulation of Fas-associated death domain-like IL-1- converting enzyme-like inhibitory protein [FLIP; long (FLIPL) and short (FLIPS) forms] in human ovarian epithelial cancer cells by TNF{alpha} and their significance in the resistance of the cells to the proapoptotic action of the cytokine. OV2008, A2780-s, and OVCAR-3 cells were cultured in serum-free media with or without cycloheximide (CHX, 10 µg/ml) ± TNF{alpha} (5, 10, 20 ng/ml) or transfected with a mammalian expression vector containing either a dominant negative inhibitor {kappa}B (I{kappa}B), FLIPS sense or antisense cDNA and cultured with or without TNF{alpha}. In the presence of CHX, TNF{alpha} increased caspase-8 and -3 cleavage and apoptosis. It also induced I{kappa}B phosphorylation, nuclear factor {kappa}B activation, and the expression of FLIPS but not of FLIPL. Overexpression of dominant negative I{kappa}B attenuated TNF{alpha}-induced FLIPS expression and enhanced TNF{alpha}-induced apoptosis. Apoptosis induced by TNF{alpha} and CHX was facilitated by FLIPS antisense expression but attenuated by sense transfection. This study demonstrates that TNF{alpha} up-regulates FLIPS expression, and this effect is mediated by the activation of nuclear factor {kappa}B. The induction of FLIPS expression by TNF{alpha} might contribute to the resistance of ovarian epithelial cancer cells to the proapoptotic action of the cytokine.




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