title: Changes in Cell Cycle and Up-Regulation of Neuronal Markers During SH-SY5Y Neurodifferentiation by Retinoic Acid are Mediated by Reactive Species Production and Oxidative Stress.
abstract: Human neuroblastoma SH-SY5Y cells have been used as an in vitro model for neurodegenerative disorders such as Parkinson's disease and can be induced to a mature neuronal phenotype through retinoic acid (RA) differentiation. However, mechanisms of RA-induced differentiation remain unclear. Here, we investigate the role of reactive species (RS) on SH-SY5Y neuroblastoma cells under RA differentiation, using the antioxidant Trolox® as co-treatment. We found that RA treatment for 7 days reduced the cell number and proliferative capacity and induced the expression of adult catecholaminergic/neuronal markers such as tyrosine hydroxylase (TH), β-III tubulin, and enolase-2. Evaluation of intracellular RS production by DCFH oxidation assay and quantification of cell non-enzymatic antioxidant activity by TRAP demonstrated that RA increases RS production. Furthermore, mitochondrial NADH oxidation showed to be inhibited under differentiation with RA. Cells subjected to co-treatment with antioxidant Trolox® demonstrated a remaining proliferative capacity and a decrease in the pro-oxidant state and RS production. Besides, antioxidant treatment restores the mitochondrial NADH oxidation. Importantly, Trolox® co-treatment inhibited the appearance of morphological characteristics such as neurite extension and branching, and decreased the expression of TH, β-III tubulin, and enolase-2 after a seven-day differentiation with RA, indicating that RS production is a necessary step in this process. Trolox® also inhibited the phosphorylation of Akt and ERK1/2, which are involved in differentiation and survival, respectively, of these cells. Altogether, these data indicate the presence of a redox-dependent mechanism in SH-SY5Y RA-differentiation process and can be a useful insight to improve understanding of neuronal differentiation signaling.
authors: Departamento de Bioquímica, Centro de Estudos em Estresse Oxidativo, Universidade Federal do Rio Grande do Sul-UFRGS, Rua Ramiro Barcelos, 2600 - Anexo, Porto Alegre, RS, CEP 90035-003, Brazil. alice.bio@hotmail.com.
authors: Alice
authors: http://orcid.org/0000-0003-2715-1215
authors: A
authors: Kunzler
authors: Departamento de Bioquímica, Centro de Estudos em Estresse Oxidativo, Universidade Federal do Rio Grande do Sul-UFRGS, Rua Ramiro Barcelos, 2600 - Anexo, Porto Alegre, RS, CEP 90035-003, Brazil.
authors: Fares
authors: F
authors: Zeidán-Chuliá
authors: Departamento de Bioquímica, Centro de Estudos em Estresse Oxidativo, Universidade Federal do Rio Grande do Sul-UFRGS, Rua Ramiro Barcelos, 2600 - Anexo, Porto Alegre, RS, CEP 90035-003, Brazil.
authors: Juciano
authors: J
authors: Gasparotto
authors: Departamento de Bioquímica, Centro de Estudos em Estresse Oxidativo, Universidade Federal do Rio Grande do Sul-UFRGS, Rua Ramiro Barcelos, 2600 - Anexo, Porto Alegre, RS, CEP 90035-003, Brazil.
authors: Carolina Saibro
authors: CS
authors: Girardi
authors: Departamento de Bioquímica, Centro de Estudos em Estresse Oxidativo, Universidade Federal do Rio Grande do Sul-UFRGS, Rua Ramiro Barcelos, 2600 - Anexo, Porto Alegre, RS, CEP 90035-003, Brazil.
authors: Karina
authors: K
authors: Klafke
authors: Departamento de Bioquímica, Centro de Estudos em Estresse Oxidativo, Universidade Federal do Rio Grande do Sul-UFRGS, Rua Ramiro Barcelos, 2600 - Anexo, Porto Alegre, RS, CEP 90035-003, Brazil.
authors: Lyvia Lintzmaier
authors: LL
authors: Petiz
authors: Departamento de Bioquímica, Centro de Estudos em Estresse Oxidativo, Universidade Federal do Rio Grande do Sul-UFRGS, Rua Ramiro Barcelos, 2600 - Anexo, Porto Alegre, RS, CEP 90035-003, Brazil.
authors: Rafael Calixto
authors: RC
authors: Bortolin
authors: Departamento de Bioquímica, Centro de Estudos em Estresse Oxidativo, Universidade Federal do Rio Grande do Sul-UFRGS, Rua Ramiro Barcelos, 2600 - Anexo, Porto Alegre, RS, CEP 90035-003, Brazil.
authors: Diana Carolina
authors: DC
authors: Rostirolla
authors: Departamento de Bioquímica, Centro de Estudos em Estresse Oxidativo, Universidade Federal do Rio Grande do Sul-UFRGS, Rua Ramiro Barcelos, 2600 - Anexo, Porto Alegre, RS, CEP 90035-003, Brazil.
authors: Alfeu
authors: A
authors: Zanotto-Filho
authors: Departamento de Bioquímica, Centro de Estudos em Estresse Oxidativo, Universidade Federal do Rio Grande do Sul-UFRGS, Rua Ramiro Barcelos, 2600 - Anexo, Porto Alegre, RS, CEP 90035-003, Brazil.
authors: Matheus Augusto
authors: MA
authors: de Bittencourt Pasquali
authors: The School of Biomedical Sciences and The Hunter Medical Research Institute, Faculty of Health, The University of Newcastle, Callaghan, Australia.
authors: Phillip
authors: P
authors: Dickson
authors: The School of Biomedical Sciences and The Hunter Medical Research Institute, Faculty of Health, The University of Newcastle, Callaghan, Australia.
authors: Peter
authors: P
authors: Dunkley
authors: Departamento de Bioquímica, Centro de Estudos em Estresse Oxidativo, Universidade Federal do Rio Grande do Sul-UFRGS, Rua Ramiro Barcelos, 2600 - Anexo, Porto Alegre, RS, CEP 90035-003, Brazil.
authors: José Cláudio Fonseca
authors: JCF
authors: Moreira
authors: Departamento de Bioquímica, Centro de Estudos em Estresse Oxidativo, Universidade Federal do Rio Grande do Sul-UFRGS, Rua Ramiro Barcelos, 2600 - Anexo, Porto Alegre, RS, CEP 90035-003, Brazil.
authors: Daniel Pens
authors: DP
authors: Gelain
mesh_terms: D000975:Antioxidants / Q000494:pharmacology; D015415:Biomarkers / Q000378:metabolism*; D002453:Cell Cycle / Q000187:drug effects*; D002454:Cell Differentiation / Q000187:drug effects*; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D048430:Cell Shape / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism; D006801:Humans; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism; D009474:Neurons / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D018384:Oxidative Stress / Q000187:drug effects*; D010641:Phenotype; D010766:Phosphorylation / Q000187:drug effects; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism*; D014212:Tretinoin / Q000494:pharmacology*; D015854:Up-Regulation / Q000187:drug effects*
chemical_list: D000975:Antioxidants; D015415:Biomarkers; D017382:Reactive Oxygen Species; D014212:Tretinoin; D051057:Proto-Oncogene Proteins c-akt; D048049:Extracellular Signal-Regulated MAP Kinases
keywords: Neuronal differentiation; Oxidative stress; Retinoic acid; SH-SY5Y; Tyrosine hydroxylase