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Tumor necrosis factor-related apoptosis inducing ligand (TRAIL) up-regulates death receptor 5 (DR5) mediated by NFκB activation in epithelial derived cell lines

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Abstract

Tumor necrosis factor-related apoptosis inducing ligand (TRAIL/APO-2L) activates nuclear factor κB (NFκB). This activation is regulated by the recruitment of an adaptor protein Fas associating death domain (FADD) to TRAIL death receptors, death receptor 4 (DR4, TRAIL-R1) and death receptor 5 (DR5 TRAIL-R2). This leads to recruitment of caspase 8 and receptor interacting protein (RIP) to the receptor complex. Upon recruitment of caspase 8 and RIP, NFκB inducing kinase (NIK) becomes activated causing NFκB activation. The role of TRAIL induced NFκB activation in epithelial cells is unknown. Herein we demonstrate that TRAIL increases expression of DR5 in human embryonic kidney (HEK) 293, MCF-7 and MDA MB 231 epithelial cell lines while DR4 expression remains unchanged. Blockage of NFκB activation either by expression of dominant negative IκB or treatment with proteasome inhibitor lactacystin eliminates TRAIL induced DR5 expression. Expression of FADD dominant negative in HEK 293 cells that prevents the recruitment of caspase 8 and RIP to TRAIL death receptors also eliminates this increase. By over expression of the p65 subunit of NFκB that increases NFκB transcriptional activity, DR5 expression was increased compared to vector alone expressing cells. By blocking TRAIL induced NFκB activation, the sensitivity of cells to undergo TRAIL induced apoptosis was significantly decreased. Conversely, the amount of TRAIL induced apoptosis was increased in HEK 293 cells over expressing p65 subunit of NFκB. Finally blockage of NFκB activation eliminates the synergistic apoptotic response of TRAIL and etoposide. Thus, TRAIL mediated NFκB activation increases DR5 expression thereby amplifying the apoptotic response of TRAIL in epithelial derived cells.

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Authors and Affiliations

  1. Manitoba Institute of Cell Biology, University of Manitoba, 675 McDermot Ave, Winnipeg, MB, R3E 0V9

    S. Shetty, J. Brown Gladden, E. S. Henson, X. Hu, J. Villanueva, N. Haney & S. B. Gibson

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  1. S. Shetty
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  2. J. Brown Gladden
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  3. E. S. Henson
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  4. X. Hu
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  7. S. B. Gibson
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Shetty, S., Gladden, J.B., Henson, E.S. et al. Tumor necrosis factor-related apoptosis inducing ligand (TRAIL) up-regulates death receptor 5 (DR5) mediated by NFκB activation in epithelial derived cell lines. Apoptosis 7, 413–420 (2002). https://doi.org/10.1023/A:1020031023947

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