Dibutyryl cyclic AMP induces differentiation of human neuroblastoma SH-SY5Y cells into a noradrenergic phenotype
Dibutyryl cyclic AMP induces differentiation of human neuroblastoma SH-SY5Y cells into a noradrenergic phenotype
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DOI：
10.1016/j.neulet.2008.07.079
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发表时间：
2008-10-10
影响因子：
2.5
通讯作者：
Akaike, Akinori
中科院分区：
文献类型：
--
作者：
Kume, Toshiaki;Kawato, Yuka;Akaike, Akinori
二丁酰环 AMP (dbcAMP) 和视黄酸 (RA) 已被证明是人儿茶酚胺能神经母细胞瘤细胞系 SH-SY5Y 细胞形态分化的诱导剂。然而，目前尚不清楚通过这些化合物进行形态分化的 SH-SY5Y 细胞是否获得儿茶酚胺能特性。我们重点关注酪氨酸羟化酶（TH）表达和细胞内去甲肾上腺素（NA）含量的变化作为功能分化的指标。使用 SH-SY5Y 细胞中的免疫细胞化学分析，用 dbcAMP (1 mM) 和 RA (10 muM) 处理三天可诱导形态变化和 TH 阳性细胞增加。 dbcAMP(1 mM)处理中TH表达细胞的百分比大于RA(10 p,M)处理中的表达细胞的百分比。此外，dbcAMP 增加了细胞内 NA 含量，而 RA 则没有。 dbcAMP 诱导的 TH 表达细胞的增加受到 KT5720（一种蛋白激酶 A (PKA) 抑制剂）的部分抑制。我们还研究了丁酸盐对SH-SY5Y细胞的影响，因为dbcAMP被细胞内酯酶酶促降解，从而导致丁酸盐的形成。丁酸盐在比 dbcAMP 更低的浓度下诱导 NA 含量增加，尽管丁酸盐对表达 TH 的细胞的增加小于 dbcAMP 的增加。 dbcAMP (1 mM) 和丁酸盐 (0.3 mM) 诱导的 NA 含量增加被 TH 抑制剂 α-甲基-p-酪氨酸 (1 mM) 完全抑制。这些结果表明 dbcAMP 通过 PKA 激活和丁酸盐诱导分化为去甲肾上腺素能表型。 (C) 2008 Elsevier Ireland Ltd. 保留所有权利。
Dibutyryl cyclic AMP (dbcAMP) and retinoic acid (RA) have been demonstrated to be the inducers of morphological differentiation in SH-SY5Y cells, a human catecholaminergic neuroblastoma cell line. However, it remains unclear whether morphologically differentiated SH-SY5Y cells by these compounds acquire catecholaminergic properties. We focused on the alteration of tyrosine hydroxylase (TH) expression and intracellular content of noradrenaline (NA) as the indicators of functional differentiation. Three days treatment with dbcAMP (1 mM) and RA (10 mu M) induced morphological changes and an increase of TH-positive cells using immunocytochemical analysis in SH-SY5Y cells. The percentage of TH-expressing cells in dbcAMP(1 mM) treatment was larger than that in RA (10 p,M) treatment. In addition, dbcAMP increased intracellular NA content, whereas RA did not. The dbcAMP-induced increase in TH-expressing cells is partially inhibited by KT5720, a protein kinase A (PKA) inhibitor. We also investigated the effect of butyrate on SH-SY5Y cells, because dbcAMP is enzymatically degraded by intracellular esterase, thereby resulting in the formation of butyrate. Butyrate induced the increase of NA content at lower concentrations than dbcAMP, although the increase in TH-expressing cells by butyrate was smaller than that by dbcAMP. The dbcAMP (1 mM)- and butyrate (0.3 mM)-induced increase in NA content was completely suppressed by alpha-methyl-p-tyrosine (1 mM), an inhibitor of TH. These results suggest that dbcAMP induces differentiation into the noradrenergic phenotype through both PKA activation and butyrate. (C) 2008 Elsevier Ireland Ltd. All rights reserved.