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Neurotrophin Effect On The SH-SY5Y Human Neuroblastoma (HN) MODEL Of Cddp-Induced Neurotoxicity

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Neurotrophins (NGF, BDNF, NT-3, NT-4/5) have several pharmacological effects on adult neurons, and it has been proposed that they may play a role in sustaining the reparative mechanisms following peripheral nerve injuries. In this study we evaluated the effect of recombinant human (rh) neurotrophins (rhNGF, gifted by Boehringer Mannheim, Germany; rhBDNF and rhNT-3 gifted by Amgen Co, USA) on cisplatin (CDDP)-induced neurotoxicity in HN SH-SY5Y neurotoxicity model. The cells were differentiated with RA 10 μM and exposed for 4 hours to 2.5 μg/ml CDDP with 50 ng/ml rhNGF, rhBDNF or rhNT-3. Then the cells were maintained in DMEM, 10 μM RA and 50 ng/ml rhNGF, rhBDNF or rhNT-3 and assessed for neurotoxicity 48, 72 and 96 hours after the first treatment with RA. Cells were considered “differentiated” when their neurite was longer than 50 μm. At each time point the neurite length was measured on at least 200 randomly selected cells/dish and the percentage of differentiated cells per culture was calculated. Exposure of RA-differentiated SH-SY5Y cells to CDDP 2.5 μg/ml induced a reduction of the neurite length and of the percentage of differentiated cells. The co-treatment of RA-differentiated SH-SY5Y neuroblastoma cells with CDDP and rhBDNF restored the neurite length and the percentage of differentiated cells to values similar to those of cells treated with RA 10 μM alone. On the contrary, exposure of RA-differentiated SH-SY5Y neuroblastoma cells to rhNT-3/CDDP combination did not significantly modify the neurite length nor the percentage of differentiated cells in comparison to the cells treated with CDDP alone. The effect of rhNGF was intermediate between that of rhBDNF and the absence of any protection observed with rhNT-3. Our results are in agreement with the observation that RA treatment induces the expression of functional trkB receptors in SH-SY5Y cells, which results in BDNF-mediated neuronal differentiation of SH-SY5Y cells and explains the BDNF neuroprotective effect observed, while trkA receptors are expressed at a lower level and trkC receptors are not expressed even after RA-induced differentiation.
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Document Type: Abstract

Affiliations: Istituto di Anatomia Umana, Università di Milano, L.I.T.A., Segrate, MI

Publication date: March 1, 2001

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