Gene Expression Profile of NF-κB, Nrf2, Glycolytic, and p53 Pathways During the SH-SY5Y Neuronal Differentiation Mediated by Retinoic Acid

Abstract

SH-SY5Y cells, a neuroblastoma cell line that is a well-established model system to study the initial phases of neuronal differentiation, have been used in studies to elucidate the mechanisms of neuronal differentiation. In the present study, we investigated alterations of gene expression in SH-SY5Y cells during neuronal differentiation mediated by retinoic acid (RA) treatment. We evaluated important pathways involving nuclear factor kappa B (NF-κB), nuclear E2-related factor 2 (Nrf2), glycolytic, and p53 during neuronal differentiation. We also investigated the involvement of reactive oxygen species (ROS) in modulating the gene expression profile of those pathways by antioxidant co-treatment with Trolox®, a hydrophilic analogue of α-tocopherol. We found that RA treatment increases levels of gene expression of NF-κB, glycolytic, and antioxidant pathway genes during neuronal differentiation of SH-SY5Y cells. We also found that ROS production induced by RA treatment in SH-SY5Y cells is involved in gene expression profile alterations, chiefly in NF-κB, and glycolytic pathways. Antioxidant co-treatment with Trolox® reversed the effects mediated by RA NF-κB, and glycolytic pathways gene expression. Interestingly, co-treatment with Trolox® did not reverse the effects in antioxidant gene expression mediated by RA in SH-SY5Y. To confirm neuronal differentiation, we quantified endogenous levels of tyrosine hydroxylase, a recognized marker of neuronal differentiation. Our data suggest that during neuronal differentiation mediated by RA, changes in profile gene expression of important pathways occur. These alterations are in part mediated by ROS production. Therefore, our results reinforce the importance in understanding the mechanism by which RA induces neuronal differentiation in SH-SY5Y cells, principally due this model being commonly used as a neuronal cell model in studies of neuronal pathologies.

Nedd4-Binding Protein 1 and TNFAIP3-Interacting Protein 1 Control MHC-1 Display in Neuroblastoma

“…For example, NF-kB activation is required for certain cytotoxic agents to kill neuroblastoma cells (45,46). Also, efficacy of retinoic acid-induced differentiation of neuroblastoma cells requires an active NF-kB pathway (47)(48)(49).…”

Retracted: Effects of Di‐isononyl Phthalate on Neuropeptide Y Expression in Differentiating Human Neuronal Cells

“…Thus, the differentiation promoting agents were added in the presence of continuous phthalate exposure. SH-SY5Y has a catecholaminergic phenotype resembling immature sympathetic neuroblasts in culture [17], and is a well-established model system to study initial stages of neuronal differentiation [18,19]. Treatment with ATRA is often used to obtain a more cholinergic phenotype at the expense of the catecholaminergic phenotype [20], and it has frequently been used as a model of dopaminergic neurons in Parkinson's disease research [21].…”

Transcriptomic Profiling Discloses Molecular and Cellular Events Related to Neuronal Differentiation in SH-SY5Y Neuroblastoma Cells

“…It is conceivable that variations occur in the expression of sets of genes, which exert their effects in different compartments of the cell according to sequentially orchestrated movements. Recently, the functional networks and biological functions involved in SH-SY5Y differentiation were analysed by gene expression techniques (microarray and quantitative real-time PCR) focusing on both genes and miRNA expression profiles of SH-SY5Y which were differentiated using RA alone or in combination with BDNF (Batagov et al 2013;de Bittencourt Pasquali et al 2016;Goldie et al 2014;Korecka et al 2013).…”