基因表达谱芯片在筛选胃腺癌相关基因中的应用  

Screening differentially expressed genes in gastric adenocarcinoma by cDNA microarray

作　　者：陈少全[1] 陈菊祥 施靖华[1] 胡志前[1] 应康[3] 唐榕[3] 李瑶[3] 符薇 谢毅[3] 毛裕民[3] 

机构地区：[1]第二军医大学长征医院普通外科,上海200003 [2]长征医院神经外科 [3]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 [4]联合基因科技(集团)有限公司

出　　处：《第二军医大学学报》2001年第6期523-526,共4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 :应用基因表达谱芯片筛选胃腺癌相关基因 ,并对这些基因的功能进行初步分析。 方法 :按一步法抽提胃腺癌和对照胃 (正常 )组织的 RNA并纯化 m RNA ;将 12 80 0条人类基因 PCR产物按微矩阵排列点样于化学涂层的载玻片上 ,制成基因芯片 ;将等量的对照胃和胃腺癌组织 m RNA分别逆转录合成荧光分子掺入的 c DNA一链做探针 ,混合后杂交上述基因芯片。经严格洗片后扫描芯片荧光信号图像 ,计算机分析后比较两种组织中差异表达的基因。结果 :在 12 80 0条基因中 ,胃腺癌与正常胃组织间存在差异表达的基因。在所检测的 5例临床标本中均存在显著差异表达的基因共 2 7条 (0 .2 1% ) ,其中上调的 11条 (0 .0 86 % ) ,下调的 16条 (0 .12 5 % ) ,下调的基因中有 2条为新基因。结论 :微矩阵基因芯片在筛选胃腺癌发生相关基因的改变时 ,具有快速、高通量、高敏度等特点 。Objective: To search for the differentially expressed genes between gastric adenocarcinoma and normal gastric mucosa using cDNA microarray. Methods: The PCR products of 12 800 human genes were spotted on a chemical material coated glass plate in array. DNAs were fixed onto the glass plate after series of treatments. The total RNAs were isolated from the tissues, and were purified to mRNAs by Oligotex. Both mRNAs from the gastric adenocarcinoma and normal gastric mucosa were reversely transcribed to the cDNAs with the incorporation of fluorescent dUTP to prepare the hybridization probes. The mixed probes were hybridized to the cDNA microarray. After high stringent washing, the cDNA microarray was scanned for the fluorescent signals and showed differences between 2 tissues. Results: Among the 12 800 target genes, 27 genes differentially expressed in all 5 samples were identified, 11 were up regulated (0.086%) and 16 down regulated (0.125%). There were 2 novel genes among the down regulated group. Conclusion: cDNA microarray technique is effective in screening the differentially expressed genes between gastric adenocarcinoma and normal gastric mucosa. [

关 键 词：基因芯片 胃肿瘤 腺癌 基因差异表达 

分 类 号：R735.2[医药卫生—肿瘤] Q78[医药卫生—临床医学]