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Persistent inhibition of FLIPL expression by lentiviral small hairpin RNA delivery restores death-receptor-induced apoptosis in neuroblastoma cells

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Abstract

Neuroblastoma represents the most common and deadly solid tumour of childhood, which disparate biological and clinical behaviour can be explained by differential regulation of apoptosis. To understand mechanisms underlying death resistance in neuroblastoma cells, we developed small hairpin of RNA produced by lentiviral vectors as tools to selectively interfere with FLIPL, a major negative regulator of death receptor-induced apoptosis. Such tools revealed highly efficient in interfering with FLIPL expression and function as they almost completely repressed endogenous and/or exogenously overexpressed FLIPL protein and fully reversed FLIPL-mediated TRAIL resistance. Moreover, interference with endogenous FLIPL and FLIPS significantly restored FasL sensitivity in SH-EP neuroblastoma cell line. These results reveal the ability of lentivirus-mediated shRNAs to specifically and persistently interfere with FLIP expression and support involvement of FLIP in the regulation of death receptor-mediated apoptosis in neuroblastoma cells. Combining such tools with other therapeutic modalities may improve treatment of resistant tumours such as neuroblastoma.

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Abbreviations

NB::

neuroblastoma

DR::

death-receptor

FLIP::

FLICE-inhibitory protein

DISC::

death-inducing signalling complex

TRAIL::

tumour necrosis factor-related apoptosis-inducing ligand

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Authors and Affiliations

  1. Paediatric Oncology Research, Paediatric Department, University Hospital CHUV, CH-1011, Lausanne

    M. Flahaut, A. Mühlethaler-Mottet, K. Auderset, K. Balmas Bourloud & N. Gross

  2. Paediatric Surgery, Paediatric Department, University Hospital CHUV, CH-1011, Lausanne

    R. Meier & J. M. Joseph

  3. Hemato-oncology Unit, Paediatric Department, University Hospital CHUV, CH-1011, Lausanne

    M. Beck Popovic

  4. Paediatric Oncology Research, Paediatrics – CHUV, CH-1011, Lausanne

    N. Gross

Authors
  1. M. Flahaut
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  2. A. Mühlethaler-Mottet
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  3. K. Auderset
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  4. K. Balmas Bourloud
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  5. R. Meier
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  6. M. Beck Popovic
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  7. J. M. Joseph
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  8. N. Gross
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Correspondence to N. Gross.

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Flahaut, M., Mühlethaler-Mottet, A., Auderset, K. et al. Persistent inhibition of FLIPL expression by lentiviral small hairpin RNA delivery restores death-receptor-induced apoptosis in neuroblastoma cells. Apoptosis 11, 255–263 (2006). https://doi.org/10.1007/s10495-006-3435-9

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  • DOI: https://doi.org/10.1007/s10495-006-3435-9

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