title: NGF sensitizes TrkA SH-SY5Y neuroblastoma cells to TRAIL-induced apoptosis.
abstract: We report a novel pro-apoptotic function for nerve growth factor (NGF) and its tropomyosin-related kinase A (TrkA) receptor in sensitizing TRAIL (TNF-related apoptotis-inducing ligand)-resistant SH-SY5Y neuroblastoma (NB) cells to TRAIL-induced apoptosis, resulting in the abrogation of anchorage-independent tumourigenic growth in vitro. We show that the TRAIL-resistant SH-SY5Y phenotype is cFLIP (cellular FLICE-like inhibitory protein) dependent and not due to low-level functional TRAIL receptor or caspase expression or an inhibitory equilibrium between functional and decoy TRAIL receptors or B-cell lymphoma 2 (Bcl-2) and BH3-only (Bcl-2 homology domain 3-only) family proteins. NGF sensitization of SH-SY5Y cells to TRAIL-induced apoptosis was dependent upon TrkA expression, activation and subsequent sequestration of cFLIP. This reduces cFLIP recruitment to TRAIL-activated death receptors and increases the recruitment of caspase-8, leading to TRAIL-induced, caspase-dependent, type II apoptosis via the intrinsic mitochondrial pathway. This effect was temporary, inhibited within 6 h by nuclear factor-κ binding (NF-κB)-mediated increase in myeloid cell leukaemia-1 (Mcl-1) expression, abrogated by transient cFLIP or B-cell lymphoma-extra large (Bcl-xL) overexpression and optimized by NF-κB and Mcl-1 inhibitors. This novel mechanism adds an important pro-apoptotic immunological dimension to NGF/TrkA interaction that may not only help to explain the association between TrkA expression, better prognosis and spontaneous remission in NB, but also provides a novel potential pro-apoptotic therapeutic use for NGF, TRAIL and inhibitors of NF-κB and/or Mcl-1 in favourable and unfavourable NBs that express TrkA and exhibit cFLIP-mediated TRAIL resistance.
authors.affiliation: Department of Applied Clinical and Biotechnological Sciences, University of L'Aquila , Via Vetoio, Coppito 2, L'Aquila 67100, Italy.
authors.forename: P
authors.initials: P
authors.lastname: Ruggeri
authors.affiliation: Department of Applied Clinical and Biotechnological Sciences, University of L'Aquila , Via Vetoio, Coppito 2, L'Aquila 67100, Italy.
authors.forename: L
authors.initials: L
authors.lastname: Cappabianca
authors.affiliation: Department of Applied Clinical and Biotechnological Sciences, University of L'Aquila , Via Vetoio, Coppito 2, L'Aquila 67100, Italy.
authors.forename: A R
authors.initials: AR
authors.lastname: Farina
authors.affiliation: Department of Applied Clinical and Biotechnological Sciences, University of L'Aquila , Via Vetoio, Coppito 2, L'Aquila 67100, Italy.
authors.forename: L
authors.initials: L
authors.lastname: Gneo
authors.affiliation: Department of Applied Clinical and Biotechnological Sciences, University of L'Aquila , Via Vetoio, Coppito 2, L'Aquila 67100, Italy.
authors.forename: A R
authors.initials: AR
authors.lastname: Mackay