Abstract
Melanoma cells are relatively resistant to Apo2L/TRAIL (TNF-related apoptosis-inducing ligand). We postulated that resistance might result from higher expression of inhibitors of apoptosis including Bcl-2, FLIP (FLICE-like inhibitory protein) or IAPs such as XIAP (X-linked inhibitor of apoptosis) or survivin. Compared to scrambled or mismatch controls, targeting individual inhibitors with siRNA (si-Bcl-2, si-XIAP, si-FLIP or si-Surv), followed by Apo2L/TRAIL resulted in marked increase in apoptosis in melanoma cells. Compared to Bcl-2 or FLIP, siRNAs against XIAP and survivin were most potent in sensitizing melanoma cells. A similar substantial increase in apoptosis was seen in renal carcinoma cells (SKRC-45, Caki-2), following the inhibition of either XIAP or survivin by siRNAs. Apo2L/TRAIL treatment in IAP-targeted cells resulted in cleavage of Bid, activation of caspase-9 and cleavage of PARP (poly ADP-ribose polymerase). Thus, Apo2L/TRAIL resistance can be overcome by interfering with expression of inhibitors of apoptosis regulating both extrinsic (death receptor) or intrinsic (mitochondrial) pathways of apoptosis in melanoma cells.
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Abbreviations
- IAP:
-
inhibitor of apoptosis
- XIAP:
-
X-linked inhibitor of apoptosis
- FLIP:
-
flice-like inhibitory protein
- siRNA:
-
small interfering ribonucleic acid
- Apo2L/TRAIL:
-
TNF-related apoptosis-inducing ligand
- si-Scr:
-
random scrambled siRNA
- IFN:
-
interferon
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Acknowledgements
We thank Dr. Avi Ashkenazi (Genentech, Inc., San Franscisco, CA, USA) for providing recombinant soluble Apo2L/TRAIL protein. We gratefully acknowledge the technical assistance by Alex Rodriguez for flow cytometry. This study was supported by NIH R01 Grant CA 90914 (EC Borden).
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Chawla-Sarkar, M., Bae, S., Reu, F. et al. Downregulation of Bcl-2, FLIP or IAPs (XIAP and survivin) by siRNAs sensitizes resistant melanoma cells to Apo2L/TRAIL-induced apoptosis. Cell Death Differ 11, 915–923 (2004). https://doi.org/10.1038/sj.cdd.4401416
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DOI: https://doi.org/10.1038/sj.cdd.4401416


