人BTBD_(10)新基因克隆及组织表达和在脑胶质瘤中的表达谱及聚类  

Cloning the novel BTBD_(10) gene and cluster analysis of gene expression in normal tissue and glioma

作　　者：陈菊祥[1] 卢亦成[1] 骆纯[1] 胡国汉[1] 楼美清[1] 徐坚[2] 应康[2] 谢毅[2] 

机构地区：[1]第二军医大学附属长征医院神经外科,上海神经外科研究所,200003 [2]复旦大学生命科学院遗传工程国家重点实验室

出　　处：《中华神经外科杂志》2004年第3期187-190,共4页Chinese Journal of Neurosurgery

基　　金：上海市科委基金资助项目(034119835);国家自然科学基金资助项目(30100192)

摘　　要：目的探讨人脑胶质瘤相关BTBD10新基因克隆、正常成人组织分布及在不同级别星形细胞瘤中的表达。方法构建胎脑cDNA文库和大规模测序获得全长BTBD10新基因;用Clontech人多组织文库(MTC)为模板研究BTBD10在8种正常组织中的分布;用含13 939种人类基因(8 347种已知基因,5 592种未知基因)的BioStarH140S型表达谱芯片检测BTBD10在星形细胞瘤中的表达,Hierarchical聚类分析与BTBD10表达谱相近的基因群;Northern杂交验证BTBD10在不同级别胶质瘤中的表达。结果人胎脑文库中克隆的BTBD10新基因含1 428bp长的开放阅读框,编码475个氨基酸的蛋白;芯片结果提示BTBD10基因在18例胶质瘤组织中表达一致降低,与促性腺诱导素转录阻抑蛋白GIOT1、血管性肠肽VIP等7条基因的表达谱相似;BTBD10在正常脑组织中表达量最高,而心、肺、肝、肾、胰腺中表达较低;Northern显示BTBD10与胶质瘤发生密切相关,与芯片结果一致。结论表达谱芯片可快速高效筛选脑胶质瘤相关基因,BTBD10基因与胶质瘤发生密切相关,在脑胶质瘤的转录调控中起重要作用。Objective To clone and identify the novel BTBD10 gene, and study the expression pattern of this gene in normal tissue and in all grades of human glioma. Methods From a fetal brain cDNA library, we isolated the novel BTBD10 gene. The expression pattern of BTBD10 gene in 8 normal tissues was examined by MTC panel (Clontech). BioStarH140S microarray (including 8 347 old genes and 5 592 novel genes) were adopted and hybridized with BTBD10 probes. Those similar expressed genes with BTBD10 were studied by Hierarchical method and Northern hybridization was used for verifying expression of BTBD10 gene in glioma. Results We isolated a cDNA of 2 489 bp encoding a novel human BTB domain containing an open reading frame (ORF) of 1 428 bp and a putative 475 amino acids protein. cDNA microarray analysis showed BTBD10 was down regulated in all 18 glioma samples and expressed similarly with GIOT1?VIP?TP53BP1, etc. The results of MTC panel showed a ubiquitous expression pattern which highly expressed in adult brain while weak expression in heart, lung, liver, kidney and pancreas. It was testified by Northern-blot and was in accordance with microarray. Conclusion cDNA microarray technology is a powerful technique in screening differentially expressed genes, BTBD10 was closely related to human glioma and might play an important role in regulating transcription of glioma.

关 键 词：BTBD10 基因克隆 组织表达 脑胶质瘤 表达谱 聚类 

分 类 号：R739.4[医药卫生—肿瘤]