Analysis of the Catecholaminergic Phenotype in Human SH-SY5Y and BE(2)-M17 Neuroblastoma Cell Lines upon Differentiation

Abstract

Human cell lines are often used to investigate cellular pathways relevant for physiological or pathological processes or to evaluate cell toxicity or protection induced by different compounds, including potential drugs. In this study, we analyzed and compared the differentiating activities of three agents (retinoic acid, staurosporine and 12-O-tetradecanoylphorbol-13-acetate) on the human neuroblastoma SH-SY5Y and BE(2)-M17 cell lines; the first cell line is largely used in the field of neuroscience, while the second is still poorly characterized. After evaluating their effects in terms of cell proliferation and morphology, we investigated their catecholaminergic properties by assessing the expression profiles of the major genes involved in catecholamine synthesis and storage and the cellular concentrations of the neurotransmitters dopamine and noradrenaline. Our results demonstrate that the two cell lines possess similar abilities to differentiate and acquire a neuron-like morphology. The most evident effects in SH-SY5Y cells were observed in the presence of staurosporine, while in BE(2)-M17 cells, retinoic acid induced the strongest effects. Undifferentiated SH-SY5Y and BE(2)-M17 cells are characterized by the production of both NA and DA, but their levels are considerably higher in BE(2)-M17 cells. Moreover, the NAergic phenotype appears to be more pronounced in SH-SY5Y cells, while BE(2)-M17 cells have a more prominent DAergic phenotype. Finally, the catecholamine concentration strongly increases upon differentiation induced by staurosporine in both cell lines. In conclusion, in this work the catecholaminergic phenotype of the human BE(2)-M17 cell line upon differentiation was characterized for the first time. Our data suggest that SH-SY5Y and BE(2)-M17 represent two alternative cell models for the neuroscience field.

RA Differentiation Enhances Dopaminergic Features, Changes Redox Parameters, and Increases Dopamine Transporter Dependency in 6-Hydroxydopamine-Induced Neurotoxicity in SH-SY5Y Cells

“…On the other hand, many lines of evidence showed that RA-differentiated cells increase their expression of these dopaminergic markers, such as TH and DAT (Påhlman et al 1984;Lopes et al 2010;Filograna et al 2015). These discrepancies in the literature might be attributable to the varying differentiation protocols used, since there are differences between them, such as duration, cell 20 densities, serum concentration and differentiation agent (e.g.…”

“…These discrepancies in the literature might be attributable to the varying differentiation protocols used, since there are differences between them, such as duration, cell 20 densities, serum concentration and differentiation agent (e.g. RA, staurosporine, BDNF) (Cheung et al 2009;Agholme et al 2010;Lopes et al 2010;Filograna et al 2015).…”

Hydrogen peroxide modifies both activity and isoforms of acetylcholinesterase in human neuroblastoma SH-SY5Y cells

“…Despite all of these works, no link between oxidative stress and AChE has been identified in human brain cells. In this context, it would be interesting to know the effect of ROS on cholinergic system markers from undifferentiated SH-SY5Y neuroblastoma cells; because, although these cells are not specifically cholinergic, they present AChE enzyme as has been previously demonstrated by Filograna et al [19]. Although there are some studies about the action of pro-oxidant substances on AChE of different cell lines, to the best of our knowledge the direct effect of oxidative stress on the levels of AChE and AChE activity in human brain cells remains unclear.…”

The SH-SY5Y cell line in Parkinson’s disease research: a systematic review

“…Intriguingly, staurosporine, a broad-spectrum kinase inhibitor, has been used in some PD-related publications to induce cell death [64–72], while other publications have used it to induce DAergic differentiation and study PD-related features [43, 44]. Early studies on SH-SY5Y cells showed differentiation towards a neuronal phenotype upon treatment with staurosporine [73, 74], which later has been characterized as catecholaminergic-like [75]. Therefore, the effects of different concentrations of staurosporine on SH-SY5Y cells should be characterized carefully to properly interpret studies that used this drug either as a differentiation agent or as an inducer of apoptosis.…”