目的:比较人胎儿脑动脉与成人脑动脉基因表达谱的差异,筛选与人胎脑动脉发育相关的差异表达基因。方法:收集3例意外流产的胎儿(胎龄18—20周)及3例成人Willis脑动脉环及主要分支,抽提组织中的总RNA,制备成生物素标记的cRNA探针后分别与Affymetrix U133A基因芯片杂交,扫描杂交信号并用专用软件分析杂交结果,筛选出在两种组织中差异表达的基因。随机选择3个差异表达基因,用荧光定量RT—PCR验证。结果:在所检测的总共22215个基因中,胎儿脑动脉与成人脑动脉相比,表达水平相差2倍、3倍、4倍、5倍以上的基因分别有935个、302个、177个、110个;在表达水平相差5倍以上的基因中,32个基因在胎儿脑动脉中高表达,78个基因在胎儿脑动脉中低表达。基因功能分类显示:25个基因与细胞信号转导和通讯有关,16个基因与细胞外基质和细胞骨架有关,与细胞防御反应有关的基因有14个,转录因子10个,另外有45个为未分类或功能未知的基因或表达序列标签(EST)片段。结论:人胎脑动脉的正常发育受到众多基因的调控,研究这些基因的功能可能有助于认清脑动脉瘤等脑动脉发育相关性疾病的发病机制。
AIM: To investigate the difference in gene expression profile between fetal cerebral arteries and adult cerebral arteries. METHODS: The cerebral arteries of 3 aborted fetus with gestation age less than 20 weeks and 3 normal adults were collected. RNA was extracted and the difference between the gene expression profile was analysized by Affymetrix genechip. Quantitative fluorescence RT - PCR was used to validate the results of genechip. RESULTS: Among the 22 215 genes examined, compared to the adult cerebral arteries, there were 935 genes differentially expressed in the feted cerebral arteries at more than 2 - fold, 302 genes at 3 - fold, 177 at 4 - fold and 110 genes at 5 - fold. At the 5 - fold level, about 78 genes underexpressed and 32 genes overexpressed in the fetal cerebral arteries. Among the 110 genes, 25 genes involved in signal transduction and communication, 16 genes related with cell structure and motility, 14 genes attributed to cell defence, 4 genes belong to transcription factor and other 45 genes were unclassifed genes or with unknown function. CONCLUSIONS : There are a plenty of genes differentially expressed in human fetal cerebral arteries and adult cerebral arteries. To illustrate the functions of these genes will be helpful to recognize the pathogenesis of the diseases derived from the cerebral arteries such as cerebral aneurysms.