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目的 进一步了解大肠癌与正常组织间基因表达谱差异,寻找可能用于临床诊断和治的基因标志。方法 应用肿瘤基因解剖工程(CGAP)数据库中基因表达短序列分析(SAGE)数据筛查大肠癌和正常组织差异表达基因,并用 RT PCR方法进一步在大肠癌细胞株(SW1116、Lovo、HCT-8、Hce-8693)和20 例组织标本中验证。结果 在 SAGE 数据库中共分析大肠癌和正常组织短序列195 160个,发现差异基因216个。对其中17个基因进行RT PCR验证分析,在细胞株中基因检出阳性率为35.3%~76.5%,组织标本中阳性率为88.2%。对其中转化生长因子β1、蛋白酶体 26S亚单位、热休克蛋白1进行半定量 RT PCR,基因差异表达率分别为 60%、50%、35%。结论 利用 CGAP数据库中SAGE数据能快捷、可靠地筛查大肠癌差异基因表达谱,对这些差异基因进一步分析可能为临床诊治大肠癌提供基因标志。
Objective To further understand the differences in gene expression profiles between colorectal cancer and normal tissues and to search for genetic markers that may be used in clinical diagnosis and treatment. Methods The differentially expressed genes in colorectal cancer and normal tissues were screened by gene expression short sequence analysis (SAGE) in tumor gene anatomy (CGAP) database and further detected by RT-PCR in colorectal cancer cell lines (SW1116, Lovo, HCT-8, Hce-8693) and 20 tissue samples. Results A total of 195 160 short sequences of colorectal cancer and normal tissues were analyzed in the SAGE database and 216 differential genes were found. RT-PCR analysis of 17 of these genes showed that the positive rate of gene detection in cell lines was 35.3% -76.5%, and the positive rate in tissue samples was 88.2%. Semi-quantitative RT-PCR was performed on TGF-β1, 26S subunit of proteasome and heat shock protein 1. The differential expression rates of genes were 60%, 50% and 35% respectively. Conclusion SAGE data in CGAP database can be used to screen differential gene expression profiles of colorectal cancer quickly and reliably. Further analysis of these differentially expressed genes may provide genetic markers for clinical diagnosis and treatment of colorectal cancer.