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  • Original Paper
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Loss of XIAP protein expression by RNAi and antisense approaches sensitizes cancer cells to functionally diverse chemotherapeutics

Oncogene volume 23, pages 8105–8117 (2004)Cite this article

Abstract

Stable expression of short-hairpin RNAs (shRNAs) directed against the X-linked inhibitor of apoptosis (XIAP) resulted in the generation of three MDA-MB-231 cell lines (XIAP shRNA cells) with reductions in XIAP mRNA and protein levels >85% relative to MDA-MB-231 cells stably transfected with the U6 RNA polymerase III promoter alone (U6 cells). This RNA interference (RNAi) approach dramatically sensitized these cells to killing by the tumor necrosis factor-related apoptosis-inducing ligand (TRAIL). Importantly, loss of XIAP also sensitized the cells to killing by taxanes but had no additional effects on killing by carboplatin and doxorubicin. The increased sensitivity of the XIAP shRNA cells to killing by TRAIL and taxanes correlated with enhanced caspase cleavage and activation, including caspase-8, and robust processing of poly(ADP-ribose) polymerase and BID compared to U6 cells. Additionally, increasing XIAP levels by adenovirus-mediated expression protected both XIAP shRNA and U6 cells from TRAIL killing in a dose-dependent manner. The effects observed by stable RNAi with respect to TRAIL sensitization were also achieved following downregulation of XIAP in Panc-1 cells treated with a second-generation, mixed-backbone antisense oligonucleotide, AEG 35156/GEM640. These data indicate that reducing XIAP protein expression by either RNAi or antisense approaches increases cancer cell susceptibility to functionally diverse chemotherapeutic agents and supports the notion that downregulation of XIAP in vivo may synergize with disease-relevant chemotherapeutic regimes, including TRAIL and taxanes, to increase the effectiveness of antineoplastic agents.

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Acknowledgements

DCM is supported by Industrial Research Fellowship from the National Sciences and Engineering Research Council of Canada. We thank Dr Peter Liston for valuable discussions and help in setting up the cell-free caspase activity assays.

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  1. Dan C McManus: Supported by a National Sciences and Engineering Research Council of Canada (NSERC) Industrial Research Fellowship

Authors and Affiliations

  1. Aegera Oncology Inc., University of Ottawa and Children's Hospital of Eastern Ontario Research Institute, Ottawa, K1H 8L1, Canada

    Dan C McManus, Charles A Lefebvre, Gabriele Cherton-Horvat, Martine St-Jean, Jon P Durkin & Eric C LaCasse

  2. Hybridon Inc., Cambridge, 02139, MA, USA

    Ekambar R Kandimalla & Sudhir Agrawal

  3. Aegera Therapeutics Inc., Verdun, H3E 1A8, Canada

    Stephen J Morris & Jon P Durkin

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  1. Dan C McManus
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Correspondence to Eric C LaCasse.

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McManus, D., Lefebvre, C., Cherton-Horvat, G. et al. Loss of XIAP protein expression by RNAi and antisense approaches sensitizes cancer cells to functionally diverse chemotherapeutics. Oncogene 23, 8105–8117 (2004). https://doi.org/10.1038/sj.onc.1207967

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