基因表达谱芯片发现36条喉鳞癌相关基因  

Study of differentially expressed genes in laryngeal squamous cell carcinoma by cDNA microarray

作　　者：陈菊祥[1] 范静平[1] 应康[2] 孙爱华[1] 廖建春[1] 唐榕[2] 黄燕[2] 李瑶[2] 谢毅[2] 毛裕民[2] 

机构地区：[1]第二军医大学长征医院耳鼻咽喉科,上海200003 [2]复旦大学生命科学学院遗传学研究所遗传工程国家重点实验室

出　　处：《第二军医大学学报》2001年第6期519-522,T003,共5页Academic Journal of Second Military Medical University

基　　金：国家自然科学基金资助项目 (3 9770 792 )

摘　　要：目的 :研究探讨喉鳞癌发生、发展中相关基因群的表达和初步功能。 方法 :按微矩阵排列的 40 96种全长基因 PCR产物制成 Bio Door40 96型微矩阵表达谱芯片 ;采用条件优化的一步法抽提喉鳞癌及正常组织总 RNA,用 Qiagen公司 Oligotexm RNA离心柱分离纯化两种组织的 m RNA;经逆转录合成荧光分子 (Cy3/ Cy5 )掺入的 c DNA一链制备表达谱探针 ,芯片杂交和严格洗片后 ,用 Scan Array30 0 0荧光扫描仪扫描芯片荧光信号图像 ,利用计算机分析肿瘤及正常组织中差异表达的基因。对所获得的基因进行分子生物信息学分析。结果 :在 40 96种基因中 ,喉鳞状细胞癌与正常组织间存在差异表达的基因。在所检测的 4对临床标本中 ,发现有差异表达的基因 36条 (0 .88% )。生物信息学分析显示 ,该 36条差异表达基因与肿瘤的发病机制可能存在相关性。结论 :喉鳞状细胞癌的发生、发展中存在多基因表达调控的改变 。Objective: To screen for the differentially expressed genes in laryngeal squamous cell carcinoma and normal laryngeal tissue using cDNA microarray. Methods: The PCR products of 4 096 genes were spotted on a chemical material coated glass plates in array. The DNAs were then fixed on the glass plate by a serial of treatments. The total RNAs were isolated from the tissues, and then were purified to mRNAs by Oligotex. Both the mRNAs from the laryngeal squamous cell carcinoma and normal tissue were reversely transcribed to cDNAs with the incorporations of fluorescent dUTP, for preparing the hybridization probes. The mixed probes were then hybridized to the cDNA microarray. After high stringent washing, the cDNA microarray was scanned for the fluorescent signals and showed the differences between 2 tissues. Results: Among the 4 096 target genes, there were 36(0.88%) genes whose expression levels differed between the carcinoma and normal tissues in all 4 cases. Bioinformatical analysis of those genes had been performed. Conclusion: DNA microarray technology is an effective technique in screening for differentially expressed genes between 2 different kinds of tissue. Further analysis of the obtained genes will help to understand the molecular mechanism of malignant carcinoma. [

关 键 词：喉肿瘤 鳞状细胞癌 基因表达 基因芯片 

分 类 号：R739.65[医药卫生—肿瘤] Q78[医药卫生—临床医学]