胶质母细胞瘤基因表达谱及相关基因的聚类研究

Cluster analysis of gene expression and glioblastoma-associated gene by using cDNA microarray

目的 探讨人脑胶质母细胞瘤发生、发展中相关基因的表达及功能。方法 用含 13939种人类基因的BioStarH14 0S型表达谱芯片 ,以正常成人脑及不同级别胶质瘤组织总RNA制备的探针杂交芯片 ;ScanArray 4 0 0 0扫描芯片荧光信号 ,提取脑及胶质瘤组织差异基因并进行生物信息分析 ;用Hierarchical聚类对胶质瘤差异基因进行特征提取 ;用Northern杂交验证及进行初步功能研究。结果正常脑与 18例不同级别胶质瘤组织间筛选出多类差异表达基因 ,通过生物信息学和Hierarchical聚类 ,发现α 连环素、微型染色体维护蛋白 7、细胞周期素B2、FBX0 5、着丝粒蛋白F基因与胶质母细胞瘤密切相关 ,该类基因在低级别胶质瘤中表达差异不明显 ,但胶质母细胞瘤中表达明显上调。Northern杂交显示该类基因与胶质母细胞瘤密切相关 ,与芯片结果一致。结论 基因表达谱芯片可快速、高效地筛选胶质瘤相关基因 ,发现的 5条基因与胶质母细胞瘤侵袭性强密切相关 。

Objective To study function and screen differentially expressed genes in development of human glioblastoma by using cDNA microarray.Methods BioStarH140S microarray (including 8 347 old genes and 5 592 novel genes) was adopted and hybridized with probes which were prepared for total RNA. These probes were isolated from 2 cases of normal adult brain tissues and 18 cases of glioma tissues. Differentially expressed genes between normal tissues and glioma tissues were analyzed after scanning microarray with ScanArray4 000. These genes were studied with Hierarchical method and Northern hybridization was used for the function of genes. Results Among the 13 939 target genes, there were multiple differentially expressed genes and 5 genes were significantly up-regulated in glioblastoma. Two of them were testified by Northern-blot.Conclusion cDNA microarray technology is a powerful technique in screening differentially expressed genes; 5 up-regulated genes would be helpful to understand the molecular mechanism of glioblastoma. These genes were tightly related to invasion of glioblastoma and might become molecular index of prognosis of glioblastoma.