摘要
目的探究NCAM1在人类神经管畸形胚胎中的作用机制。方法提取NTDs组以及同周对照组胚胎脑组织,应用人类基因组U133Plus2.0芯片杂交,比较得出差异表达谱。所有差异表达基因按照功能分类。检测目的基因NCAM1的差异表达,并应用半定量RT-PCR方法以及免疫组化验证其mRNA及蛋白的改变。结果芯片显示出NTDs较正常对照组基因谱有较大的差异改变,总共检测到22209条基因的改变,其中2倍以上增高表达的74条,2倍以上降低表达的387条,分属于物质代谢、基因表达调控、信号转导、凋亡等途径。NCAM1呈21.8倍高表达。RT-PCR以及免疫组化也验证其mRNA及蛋白呈现高表达结果(P<0.01)。结论人类神经管畸形是不同阶段发展的复杂事件,作为细胞表面粘附分子的NCAM1在神经管畸形的发生发展过程中起了重要的作用。
Objective： To explore the pathogenesis of NCAM1 in human NTDs. Methods： The human NTDs group and age - matched normal group samples were extracted from brain tissues, each group sample was hybridized to an Affymetrix Human Genome U133 Plus2.0 Array. The differentially expression profiles were analyzed using Microarray Suite 5.0 （Affymetrix） and GeneSpring 6. 0 （Silicon Genetics）. All these target genes were classified to different pathways. We detect the expressions of NCAM1 gene by Semiquantitative RT -PCR and its protein was detected by immunohistochemistry. Results： All NTDs showed large variance in expression profiles. We obtained 22209 differentially expressed genes in human NTDs including 74 genes over - expressed above 2 times and 387 genes low - expressed above 2 times , the genes were belong to metabolism, gene translation regulator, signaling, apoptosis and so on. From the data of geneehip, NCAMlis over - expressed at 21.8. The results of Semi - quantitative RT - PCR shows that the expres- sions of NCAM1 mRNA are higher expressed in NTDs than in eontrol＇s as same as the expressions of its protein （ P 〈 0. 01 ）. Conclusion： The data suggest that the development of NTDs is a complicated course at different stages. NCAM1 as a cell surface adhesion glyeoprotein plays an important role in the development and stability of human NTDs .
出处
《中国优生与遗传杂志》
2009年第2期14-16,87,共4页
Chinese Journal of Birth Health & Heredity
