摘要
以基因表达谱芯片对人正常肝及肝癌组织基因表达的差异性进行了研究比较。将4096条人cDNA用点样仪点在特制玻片上制备成表达谱芯片;利用肝和肝癌组织的mRNA通过逆转录方法,将Cy3和Cy5 2种荧光分别标记到两种组织的cDNA上,制备成cDNA探针,并与表达谱芯片进行杂交及扫描,重复4次实验,通过计算机数据处理判定基因是否在上述2种组织中有表达差异,筛选出差异表达的基因共903条。基因芯片技术可同时定量研究数以千计的基因表达水平,从而鉴定参与肿瘤发生的基因。
Large scale gene expression profiles between normal hepatocytes and hepatocellular carcinomas are detected with high throughput microarray. DNA microarrays were made by spotting PCR products of 4096 human cDNAs onto a specially treated glass slides. The probes were prepared by labelling normal tissue mRNA and cancer tissue mRNA with Cy3-dUTP and Cy5-dUTP separately through reverse transcription. The arrays were then hybridized against the cDNA probe mixture and the fluorescent signals were scanned. The obtained data were analyzed from 4 repeated experiments. There are 903 genes of which the average ratios between Cy3 signal and Cy5 signal are greater than 3.0. These genes identified through this approach are potential candidates for hepatocellular carcinoma diagnosis and therapy.
关键词
基因
基因芯片
表达谱
肝细胞癌
gene
DNA microarray
gene expression profile
hepatocellular carcinoma