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Interferon regulatory factor-1-induced apoptosis mediated by a ligand-independent fas-associated death domain pathway in breast cancer cells

Abstract

Interferon (IFN) regulatory factor-1 (IRF-1) is a transcription factor that has apoptotic anti-tumor activity. In breast cancer cell types, IRF-1 is implicated in mediating apoptosis by both novel and established anti-tumor agents, including the anti-estrogens tamoxifen and faslodex. Here we demonstrate that in MDA468 breast cancer cells, apoptosis by IFN-γ is mediated by IRF-1 and IFN-γ, and IRF-1-induced apoptosis is caspase-mediated. IRF-1 induction results in cleavage of caspase-8, -3 and -7, and application of caspase inhibitors attenuate activated cleavage products. IRF-1-induced apoptosis involves caspase-8 since apoptosis is significantly decreased by the caspase-8-specific inhibitor IETD, c-FLIP expression and in caspase-8-deficient cancer cells. Furthermore, we demonstrate that IRF-1-induced apoptosis requires fas-associated death domain (FADD) since dominant-negative FADD expressing cells resist IRF-1-induced apoptosis and activated downstream products. Immunofluorescent studies demonstrate perinuclear colocalization of FADD and caspase-8. Despite the known role of FADD in mediating death-ligand induced apoptosis, neutralizing antibodies against classical death receptors do not inhibit IRF-1 induced apoptosis, and no secreted ligand appears to be involved since MDA468 coincubated with IRF-1 transfected cells do not apoptose. Therefore, we demonstrate that IRF-1 induces a ligand-independent FADD/caspase-8-mediated apoptosis in breast cancer cells.

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Abbreviations

EFs:

embryonic fibroblasts

FADD:

fas-associated death domain

IFN:

interferon

IRF-1:

IFN regulatory factor-1

MIS:

mullerian inhibiting substance

MOI:

multiplicity of infection

siRNA:

small interfering RNA

TNF:

tumor necrosis factor

TRAIL:

TNF-related apoptosis inducing ligand

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Acknowledgements

This work was supported by NIH Grant CA098403. We thank Dr Timothy R Billiar for review of the data and helpful suggestions and Dr Donna Stolz and the Center for Biologic Imaging for assistance with immunofluorescent studies.

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Correspondence to J H Yim.

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Supplementary Information accompanies the paper on the Oncogene website (http://www.nature.com/onc).

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Stang, M., Armstrong, M., Watson, G. et al. Interferon regulatory factor-1-induced apoptosis mediated by a ligand-independent fas-associated death domain pathway in breast cancer cells. Oncogene 26, 6420–6430 (2007). https://doi.org/10.1038/sj.onc.1210470

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