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人SH-SY5Y和BE(2)-M17神经母细胞瘤细胞系分化后儿茶酚胺能表型的分析

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

作者信息

Filograna Roberta, Civiero Laura, Ferrari Vanni, Codolo Gaia, Greggio Elisa, Bubacco Luigi, Beltramini Mariano, Bisaglia Marco

机构信息

Molecular Physiology and Biophysics Unit, Department of Biology, University of Padova, Padova, Italy.

General Pathology Unit, Department of Biology, University of Padova, Padova, Italy.

出版信息

PLoS One. 2015 Aug 28;10(8):e0136769. doi: 10.1371/journal.pone.0136769. eCollection 2015.

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.

摘要

人类细胞系常用于研究与生理或病理过程相关的细胞通路,或评估不同化合物(包括潜在药物)诱导的细胞毒性或保护作用。在本研究中,我们分析并比较了三种试剂(视黄酸、星形孢菌素和12-O-十四烷酰佛波醇-13-乙酸酯)对人神经母细胞瘤SH-SY5Y和BE(2)-M17细胞系的分化活性;第一种细胞系在神经科学领域广泛使用,而第二种细胞系的特征仍不太明确。在评估它们对细胞增殖和形态的影响后,我们通过评估参与儿茶酚胺合成和储存的主要基因的表达谱以及神经递质多巴胺和去甲肾上腺素的细胞浓度,研究了它们的儿茶酚胺能特性。我们的结果表明,这两种细胞系具有相似的分化能力,并能获得神经元样形态。在SH-SY5Y细胞中,在星形孢菌素存在的情况下观察到最明显的效果,而在BE(2)-M17细胞中,视黄酸诱导的效果最强。未分化的SH-SY5Y和BE(2)-M17细胞的特征是同时产生去甲肾上腺素和多巴胺,但它们在BE(2)-M17细胞中的水平要高得多。此外,去甲肾上腺素能表型在SH-SY5Y细胞中似乎更明显,而BE(2)-M17细胞具有更突出的多巴胺能表型。最后,在两种细胞系中,星形孢菌素诱导分化后儿茶酚胺浓度均显著增加。总之,在这项工作中,首次对人BE(2)-M17细胞系分化后的儿茶酚胺能表型进行了表征。我们的数据表明,SH-SY5Y和BE(2)-M17代表了神经科学领域的两种替代细胞模型。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8b0a/4552590/c2fed8133a6c/pone.0136769.g001.jpg

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