Cheung W M, Chu A H, Ip N Y
Department of Biology, The Hong Kong University of Science and Technology, Clear Water Bay.
J Neurochem. 1997 May;68(5):1882-8. doi: 10.1046/j.1471-4159.1997.68051882.x.
Retinoic acid (RA) induced the terminal differentiation of a human embryonal carcinoma cell line (NT2/D1) into several morphologically distinct cell types, including the postmitotic CNS neurons. Although RA has been suggested to play an important role in brain development, little is known about the molecular mechanism by which RA induces neuronal differentiation. In the present study, RNA fingerprinting by arbitrarily primed PCR (RAP-PCR) was used to identify the transcripts in NT2/D1 cells that were differentially regulated by RA. Northern blot analysis of the differentially amplified PCR fragments revealed 11 genes that were regulated by RA. Of these, seven were up-regulated and four were down-regulated along the course of RA treatment. More importantly, four of the RA-regulated genes that were identified in the present study are novel. Our findings suggested that there are a number of RA-regulated genes that have yet to be identified. RAP-PCR provides a useful tool for studying the patterns of transcript expression during the course of RA treatment and allows the cloning of novel genes involved in the process of neuronal differentiation. Furthermore, it provides a basis for the selection of genes that are involved in the RA-induced signaling pathway in the human CNS.
维甲酸（RA）可诱导人胚胎癌细胞系（NT2/D1）终末分化为几种形态学上不同的细胞类型，包括有丝分裂后的中枢神经系统神经元。尽管RA被认为在脑发育中起重要作用，但关于RA诱导神经元分化的分子机制却知之甚少。在本研究中，采用任意引物PCR（RAP-PCR）进行RNA指纹分析，以鉴定NT2/D1细胞中受RA差异调控的转录本。对差异扩增的PCR片段进行Northern印迹分析，结果显示有11个基因受RA调控。其中，7个基因在RA处理过程中上调，4个基因下调。更重要的是，本研究中鉴定出的4个受RA调控的基因是新发现的。我们的研究结果表明，还有许多受RA调控的基因有待鉴定。RAP-PCR为研究RA处理过程中转录本表达模式提供了一个有用的工具，并能克隆参与神经元分化过程的新基因。此外，它为选择参与人中枢神经系统RA诱导信号通路的基因提供了基础。
