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首页> 外文期刊>Life sciences >DIFFERENTIATION AGENTS ENHANCE CHOLINERGIC CHARACTERISTICS OF LA-N-2 HUMAN NEUROBLASTOMA CELLS
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DIFFERENTIATION AGENTS ENHANCE CHOLINERGIC CHARACTERISTICS OF LA-N-2 HUMAN NEUROBLASTOMA CELLS

机译：分化剂增强LA-N-2人类神经母细胞瘤细胞的胆碱特性

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摘要

LA-N-2, a cell line derived from a human peripheral neuroblastoma, has a partially cholinergic phenotype and is a potential in vitro model of cholinergic neurons. The object of this study was to enhance the cholinergic phenotype of these cells with differentiation agents to improve the cell line's usefulness as a convenient model of cholinergic function. I treated cells in the presence of serum with 10 mu M 5-azacytidine, 2.5 mu M bromodeoxyuridine, 2 nM ciliary neurotrophic factor, 1 mM dibutyryl cAMP, 0.25 nM leukemia inhibitory factor and/or 3.8 nM nerve growth factor, N-2 supplement (without serum), or 10 mu M retinoic acid for 9-14 days. Treated cells were loaded with [H-3]choline for 30 min at 37 degrees and washed. The amounts of cellular and released (5 min, room temperature), labeled and unlabeled acetylcholine and choline were determined by HPLC. None of the differentiation agents induced Ca2+-dependent release of [H-3]acetylcholine, but 5-azacytidine, dibutyryl cAMP, N-2, and retinoic acid increased Ca2+-independent release that was specific for acetylcholine. In addition, 5-azacytidine, bromodeoxyuridine, leukemia inhibitory factor, and N-2 substantially increased [H-3]acetylcholine levels, and these increases correlated highly with increases in total acetylcholine levels. Overall, LA-N-2 cells should prove to be a good model for studying cholinergic function. [References: 12]
机译：LA-N-2是一种源自人周围神经母细胞瘤的细胞系，具有部分胆碱能表型，是胆碱能神经元的潜在体外模型。这项研究的目的是用分化剂增强这些细胞的胆碱能表型，以提高细胞系作为胆碱能功能的便利模型的有用性。我在存在血清的情况下用10μM5-氮杂胞苷，2.5μM溴脱氧尿苷，2 nM睫状神经营养因子，1 mM二丁酰cAMP，0.25 nM白血病抑制因子和/或3.8 nM神经生长因子，N-2补充剂处理了细胞（不含血清）或10μM视黄酸（9-14天）。将处理过的细胞在37度下用[H-3]胆碱加载30分钟，然后洗涤。通过HPLC测定细胞和释放的量（5分钟，室温），标记的和未标记的乙酰胆碱和胆碱的量。没有一种分化剂能诱导[H-3]乙酰胆碱的Ca2 +依赖性释放，但是5-氮杂胞苷，二丁酰基cAMP，N-2和视黄酸会增加对乙酰胆碱具有特异性的Ca2 +非依赖性释放。此外，5-氮杂胞苷，溴脱氧尿苷，白血病抑制因子和N-2显着增加了[H-3]乙酰胆碱的水平，这些增加与总乙酰胆碱水平的增加高度相关。总体而言，LA-N-2细胞应被证明是研究胆碱能功能的良好模型。 [参考：12]

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