All-trans-retinoic acid inhibits collapsin response mediator protein-2 transcriptional activity during SH-SY5Y neuroblastoma cell differentiation
All-trans-retinoic acid inhibits collapsin response mediator protein-2 transcriptional activity during SH-SY5Y neuroblastoma cell differentiation
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DOI：
10.1111/j.1742-4658.2006.05597.x
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发表时间：
2007-01-01
期刊：
影响因子：
5.4
通讯作者：
Rouzaut, Ana
中科院分区：
文献类型：
--
作者：
Fontan-Gabas, Lorena;Oliemuller, Erik;Rouzaut, Ana
神经元是高度极化的细胞，由两个在结构和功能上不同的部分组成，即轴突和树突。这种不对称结构的建立是一个受到严格调控的过程。事实上，参与微管晶格构象的蛋白质的改变在神经肿瘤疾病中很常见。这些细胞质介质之一是被称为 collapsin反应调节蛋白 - 2（CRMP - 2）的蛋白质，它与微管蛋白相互作用并促进微管蛋白聚合。在这项研究中，我们研究了全反式维甲酸诱导的SH - SY5Y神经母细胞瘤细胞分化过程中CRMP - 2的转录调控。全反式维甲酸被认为是一种潜在的预防和治疗药物，并已广泛用于体外分化神经母细胞瘤细胞。因此，我们首先证明了在分化过程中CRMP - 2的mRNA水平下调。在完成缺失构建体分析、诱变以及迁移率变动分析后，我们得出结论：CRMP - 2的基础启动子活性受转录因子AP - 2和Pax - 3调控，而E2F、Sp1和NeuroD1似乎不参与其调控。此外，我们最终确定，全反式维甲酸暴露后CRMP - 2表达降低与Pax - 3和AP - 2与其在CRMP - 2启动子中的共有序列结合受损有关。AP - 2结合减少是其在细胞质中积累的结果。另一方面，由于全反式维甲酸介导的转录抑制，Pax - 3显示出较低的结合。揭示全反式维甲酸对神经母细胞瘤细胞作用背后的分子机制很可能为其临床应用提供新的视角。
Neurons are highly polarized cells composed of two structurally and functionally distinct parts, the axon and the dendrite. The establishment of this asymmetric structure is a tightly regulated process. In fact, alterations in the proteins involved in the configuration of the microtubule lattice are frequent in neuro-oncologic diseases. One of these cytoplasmic mediators is the protein known as collapsin response mediator protein-2, which interacts with and promotes tubulin polymerization. In this study, we investigated collapsin response mediator protein-2 transcriptional regulation during all-trans-retinoic acid-induced differentiation of SH-SY5Y neuroblastoma cells. All-trans-retinoic acid is considered to be a potential preventive and therapeutic agent, and has been extensively used to differentiate neuroblastoma cells in vitro. Therefore, we first demonstrated that collapsin response mediator protein-2 mRNA levels are downregulated during the differentiation process. After completion of deletion construct analysis and mutagenesis and mobility shift assays, we concluded that collapsin response mediator protein-2 basal promoter activity is regulated by the transcription factors AP-2 and Pax-3, whereas E2F, Sp1 and NeuroD1 seem not to participate in its regulation. Furthermore, we finally established that reduced expression of collapsin response mediator protein-2 after all-trans-retinoic acid exposure is associated with impaired Pax-3 and AP-2 binding to their consensus sequences in the collapsin response mediator protein-2 promoter. Decreased attachment of AP-2 is a consequence of its accumulation in the cytoplasm. On the other hand, Pax-3 shows lower binding due to all-trans-retinoic acid-mediated transcriptional repression. Unraveling the molecular mechanisms behind the action of all-trans-retinoic acid on neuroblastoma cells may well offer new perspectives for its clinical application.