The TrkAIII oncoprotein inhibits mitochondrial free radical ROS-induced death of SH-SY5Y neuroblastoma cells by augmenting SOD2 expression and activity at the mitochondria, within the context of a tumour stem cell-like phenotype

Pierdomenico Ruggeri; Antonietta R Farina; Natalia Di Ianni; Lucia Cappabianca; Marzia Ragone; Giulia Ianni; Alberto Gulino; Andrew R Mackay

doi:10.1371/journal.pone.0094568

The TrkAIII oncoprotein inhibits mitochondrial free radical ROS-induced death of SH-SY5Y neuroblastoma cells by augmenting SOD2 expression and activity at the mitochondria, within the context of a tumour stem cell-like phenotype

PLoS One. 2014 Apr 15;9(4):e94568. doi: 10.1371/journal.pone.0094568. eCollection 2014.

Authors

Pierdomenico Ruggeri¹, Antonietta R Farina¹, Natalia Di Ianni², Lucia Cappabianca¹, Marzia Ragone¹, Giulia Ianni¹, Alberto Gulino³, Andrew R Mackay¹

Affiliations

¹ Department of Applied Clinical and Biotechnological Sciences, University of L'Aquila, L'Aquila, Italy.
² Department of Applied Clinical and Biotechnological Sciences, University of L'Aquila, L'Aquila, Italy; Department of Medical-Surgical Science and Biotechnology, University of Rome "La Sapienza", Latina, Italy.
³ Department of Experimental Medicine, University of Rome "La Sapienza", Rome, Italy.

Abstract

The developmental and stress-regulated alternative TrkAIII splice variant of the NGF receptor TrkA is expressed by advanced stage human neuroblastomas (NBs), correlates with worse outcome in high TrkA expressing unfavourable tumours and exhibits oncogenic activity in NB models. In the present study, we report that constitutive TrkAIII expression in human SH-SY5Y NB cells inhibits Rotenone, Paraquat and LY83583-induced mitochondrial free radical reactive oxygen species (ROS)-mediated death by stimulating SOD2 expression, increasing mitochondrial SOD2 activity and attenuating mitochondrial free radical ROS production, in association with increased mitochondrial capacity to produce H2O2, within the context of a more tumour stem cell-like phenotype. This effect can be reversed by the specific TrkA tyrosine kinase inhibitor GW441756, by the multi-kinase TrkA inhibitors K252a, CEP-701 and Gö6976, which inhibit SOD2 expression, and by siRNA knockdown of SOD2 expression, which restores the sensitivity of TrkAIII expressing SH-SY5Y cells to Rotenone, Paraquat and LY83583-induced mitochondrial free radical ROS production and ROS-mediated death. The data implicate the novel TrkAIII/SOD2 axis in promoting NB resistance to mitochondrial free radical-mediated death and staminality, and suggest that the combined use of TrkAIII and/or SOD2 inhibitors together with agents that induce mitochondrial free radical ROS-mediated death could provide a therapeutic advantage that may also target the stem cell niche in high TrkA expressing unfavourable NB.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Carcinogenesis / drug effects
Cell Death / drug effects
Cell Differentiation / drug effects
Cell Line, Tumor
Cell Survival / drug effects
Disease Progression
Drug Resistance, Neoplasm
Gene Expression Regulation, Neoplastic* / drug effects
Gene Knockdown Techniques
Humans
Mitochondria / drug effects
Mitochondria / metabolism*
NF-kappa B / metabolism
Neoplastic Stem Cells / drug effects
Neoplastic Stem Cells / pathology*
Neuroblastoma / pathology*
Oncogene Proteins / antagonists & inhibitors
Oncogene Proteins / metabolism
Phenotype
Phosphotransferases (Alcohol Group Acceptor) / metabolism
Protein Isoforms / antagonists & inhibitors
Protein Isoforms / metabolism
Protein Kinase Inhibitors / pharmacology
Reactive Oxygen Species / metabolism*
Receptor, trkA / antagonists & inhibitors
Receptor, trkA / metabolism*
Spheroids, Cellular / drug effects
Spheroids, Cellular / pathology
Superoxide Dismutase / deficiency
Superoxide Dismutase / genetics*

Substances

NF-kappa B
Oncogene Proteins
Protein Isoforms
Protein Kinase Inhibitors
Reactive Oxygen Species
Superoxide Dismutase
superoxide dismutase 2
Phosphotransferases (Alcohol Group Acceptor)
Inositol 1,4,5-trisphosphate 3-kinase
Receptor, trkA

Grants and funding

This work was supported by grants from: AIRC; the Italian Ministery for University and Research “Progetti di ricerca di interesse natzionali” (PRIN); and the Maugeri Foundation. Dr. Natalia Di Ianni is a recipient of an INAIL research grant (PMS45/07). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.