应用寡核苷酸芯片筛选维甲酸诱导神经母细胞瘤细胞系SH-SY5Y分化成神经元过程中的差异表达基因

目的:应用寡核苷酸芯片筛选维甲酸(RA)诱导神经母细胞瘤细胞系SH-SY5Y分化成神经元过程中的差异表达基因。方法:从人胎脑及不同类型神经系统肿瘤组织中获取目的基因,查询相应基因mRNA序列,设计并合成探针,制备了含218种基因的神经功能相关的寡核苷酸芯片。应用RA诱导SH-SY5Y8d分化成成熟神经元,提取对照组和实验组每天的总RNA,通过逆转录荧光标记cDNA探针并与芯片杂交,洗片后扫描获取图像,数据分析获得差异表达基因,并通过RT-PCR进行验证。结果:发现13种基因表达上调,没有得到下调基因。RT-PCR验证结果基本与芯片结果一致。结论:SH-SY5Y经RA诱导分化成神经元存在一些差异表达的基因,寡核苷酸芯片技术可为研究SH-SY5Y诱导分化成神经元的分子作用机理提供技术平台。

Screening Differentially Expressed Genes in the Process of Retinoic Acid Inducing SH-SY5Y to Neuron by Oligonucleotide Microarray

Objective: To study the gene expression profile in the process of retinoic acid(RA) inducing SH-SY5Y to neuron by oligonucleotide microarray. Methods: Nerve associated genes were obtained from embryonic brain and different kinds of nervous tumors. Through GenBank, the mRNA sequence of the genes were acquired. The probes were designed and synthesized. Then oligonucleotide microarray were fabricated containing 218 genes. This microarray was used to analyze gene expression in neuroblastoma differentiation by RA from 1 to 8 days. The screened genes were identified and verified by semi-quantitative RT-PCR. Results: After analyzed, the data showed that there were 13 up-regulated genes, but no down-regulated gene. Conclusion: There are some diferentially expressed genes in the process of RA inducing SH-SY5Y to neuron. Our results indicate that the oligonucleotide microarray is reliable and can be used for dissecting molecular mechanism in the progress of neuroblastoma differentiation.