基因芯片技术在胰腺癌相关基因研究中的应用  

DNA microarray analysis of gene expression profiles in pancreatic carcinoma

作　　者：谭志军[1] 胡先贵[2] 应康[3] 李瑶[3] 唐榕[3] 曹贵松[2] 唐岩[2] 金钢[2] 

机构地区：[1]中国人民解放军第254医院普外科,天津300142 [2]第二军医大学长海医院普外科 [3]复旦大学生命科学院遗传研究所

出　　处：《上海医学》2004年第3期180-183,共4页Shanghai Medical Journal

基　　金：国家自然科学基金资助项目 ( 3 0 0 0 0 160 )

摘　　要：目的　应用基因芯片技术比较胰腺癌组织与正常胰腺组织基因表达谱的差异 ,以寻找新的诊断指标和治疗位点。方法　将 12 80 0 0个人类全长基因的PCR产物点样在特殊处理后的玻片上制备基因芯片。采用逆转录的方法分别将Cy5 dUTP标记胰腺癌组织mRNA、Cy3 dUTP标记正常胰腺组织mRNA以制备探针。将每一标本的混合探针与芯片杂交 ,用ScanArray 30 0 0荧光扫描仪扫描荧光信号。应用ImaGene3.0软件分析、计算每点的Cy5和Cy3信号值。以Cy5 /Cy3比值的自然对数绝对值 >0 .6 9为标准 ,筛选差异表达基因。结果　在所检测的 6例胰腺癌标本中 ,共有 5 4个基因有差异表达 ,其中胰腺癌组织中表达上调基因 32个 (包括基因库中已登录基因 2 4个 ) ,表达下调基因 2 2个 (包括基因库中已登录基因 15个 )。结论　基因芯片技术为阐明胰腺癌特异基因表达谱提供了强有力的工具。Objective To identify new diagnostic markers and drug targets, the gene expression profiles of pancreatic cancer were compared with those of normal pancreatic tissues, utilizing cDNA microarray analysis.Methods cDNA microarray was set up by spotting PCR products of 12 800 human genes onto a specially prepared glass slides. The cDNA probes were prepared by labeling normal and cancer tissue mRNA with Cy3-dUTP and Cy5-dUTP separately through reverse transcription. The mixed probes were then hybridized to the cDNA microarray. The chips were scanned by ScanArray 3000 laser scanner at two wavelengths. The acquired image was analyzed by ImaGene 3.0 software. The intensities of each spot at the two wavelengths represented the signal intensities of Cy3-dUTP and Cy5-dUTP, respectively, the ratio of Cy3 to Cy5 was then computed. To screen differentially expressed genes according to the criterion that the absolute value of natural logarithm of the ratio of Cy5-dUTP to Cy3-dUTP was greater than 0.69.Results Among the six samples investigated, there were 54 genes, which exhibited differential expression, including 32 over-expressed genes (24 already in Genebank), and 22 under-expressed genes (15 already in Genebank) in cancerous tissues. Conclusions Technology of cDNA microarray provides a powerful approach to elucidate tumor-specific gene expression profiles in human pancreatic carcinoma.

关 键 词：基因芯片技术 胰腺癌 基因 研究 

分 类 号：R735.9[医药卫生—肿瘤]