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Papers In Press, published online ahead of print January 5, 2001
J. Biol. Chem, 10.1074/jbc.C000569200
Submitted on August 21, 2000
Revised on January 5, 2001
Accepted on January 5, 2001
Department of Medicine, Beth Israel Deaconess Medical Center, Boston, MA 02115
Corresponding Author: jmier{at}caregroup.harvard.edu
The caspase-8 homologue Flice Inhibitory Protein (FLIP) functions as a caspase-8 dominant negative, blocking apoptosis induced by the oligomerization of the adapter protein FADD/MORT-1. FLIP expression correlates with resistance to apoptosis induced by various members of the TNF family such as TRAIL. Furthermore, forced expression of FLIP renders cells resistant to fas-mediated apoptosis. Although FLIP expression is regulated primarily by MEK1 activity in activated T cells, the oncogenic signaling pathways that regulate FLIP expression in tumor cells are largely unknown. In this report, we examined the roles of the MAP kinase and PI3-kinase signaling pathways in the regulation of FLIP expression in tumor cells. We observed that the MEK1 inhibitor PD98059 reduced FLIP levels in only two of eleven tumor cell lines tested. In contrast, disruption of the PI3-kinase pathway with the specific inhibitor LY294002 reduced Akt (pkB) phosphorylation and the levels of FLIP protein and mRNA in all cell lines evaluated. The introduction of a dominant negative Akt adenoviral construct also consistently reduced FLIP expression as well as the phosphorylation of the Akt target glycogen synthase kinase-3. In addition, infection of the same cell lines with a constitutively active Akt adenovirus increased FLIP expression and the phosphorylation of GSK-3. These data add FLIP to the growing list of apoptosis inhibitors whose expression or function are regulated by the PI3-kinase-Akt pathway.
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