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    Effects of cIAP-1, cIAP-2 and XIAP triple knockdown on prostate cancer cell susceptibility to apoptosis, cell survival and proliferation.

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    Authors
    Gill, Catherine
    Dowling, Catherine
    O'Neill, Amanda J
    Watson, R William G
    Affiliation
    UCD School of Medicine and Medical Science, University College Dublin, Dublin, Ireland. cgill@hrb.ie
    Issue Date
    2009
    MeSH
    Apoptosis
    Cell Line, Tumor
    Cell Proliferation
    Cell Survival
    Data Interpretation, Statistical
    Etoposide
    Gene Expression
    Gene Knockdown Techniques
    Humans
    Inhibitor of Apoptosis Proteins
    Male
    Peptide Hydrolases
    Prostatic Neoplasms
    RNA, Small Interfering
    TNF-Related Apoptosis-Inducing Ligand
    Tunicamycin
    X-Linked Inhibitor of Apoptosis Protein
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    Citation
    Effects of cIAP-1, cIAP-2 and XIAP triple knockdown on prostate cancer cell susceptibility to apoptosis, cell survival and proliferation. 2009, 8:39 Mol. Cancer
    Journal
    Molecular cancer
    URI
    http://hdl.handle.net/10147/94206
    DOI
    10.1186/1476-4598-8-39
    PubMed ID
    19549337
    Abstract
    BACKGROUND: Manipulating apoptotic resistance represents an important strategy for the treatment of hormone refractory prostate cancer. We hypothesised that the Inhibitor of Apoptosis (IAP) Proteins may be mediating this resistance and knockdown of cIAP-1, cIAP-2 and XIAP would increase sensitivity to apoptosis. METHODS: cIAP-1, cIAP-2 and XIAP where knocked down either individually or in combination using siRNA in androgen independent prostate cancer PC-3 cells as confirmed by real-time PCR and western blotting. Cells were then treated with TRAIL, Etoposide, or Tunicamycin, and apoptosis assessed by PI DNA staining. Apoptosis was confirmed with Annexin V labelling and measurement of PARP cleavage, and was inhibited using the pan-caspase inhibitor, zVAD.fmk. Clonogenic assays and assessment of ID-1 expression by western blotting were used to measure recovery and proliferation. RESULTS: PC-3 are resistant to TRAIL induced apoptosis and have elevated expression of cIAP-1, cIAP-2 and XIAP. Combined knockdown sensitised PC-3 to TRAIL induced apoptosis, but not to Etoposide or Tunicmycin, with corresponding increases in caspase activity and PARP cleavage which was inhibited by ZVAD.fmk. Triple knock down decreased proliferation which was confirmed by decreased ID-1 expression. CONCLUSION: Simultaneous knock down of the IAPs not only sensitised the PC-3 to TRAIL but also inhibited their proliferation rates and clonogenic survival. The inability to alter sensitivity to other triggers of apoptosis suggests that this effect is specific for death receptor pathways and knock down might facilitate immune-surveillance mechanisms to counter cancer progression and, in combination with therapeutic approaches using TRAIL, could represent an important treatment strategy.
    Language
    en
    ISSN
    1476-4598
    ae974a485f413a2113503eed53cd6c53
    10.1186/1476-4598-8-39
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