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目的筛查人大肠癌组织与相应正常大肠粘膜差异表达基因。方法应用美国AffymetrixHG-U133寡核苷酸芯片(代表迄今所知的32264个人类全基因,包括19308个已知的人类基因和12956个EST簇)检测9例大肠癌组织及相应正常大肠粘膜基因表达谱,并以实时荧光定量PCR技术对基因芯片检测结果进行验证;综合运用交集补集、秩和检验及t检验比较两组表达谱数据。结果获得大肠癌组织与正常大肠粘膜组织差异表达基因和ESTs3125个(包括肿瘤上调基因ESTs974个、下调基因ESTs2151个);大肠癌组织表达而相应正常粘膜不表达的ESTs245个;大肠癌组织不表达而正常粘膜表达的ESTs162个;最重要的大肠癌差异表达基因ESTs17个。本研究所筛得之大肠癌差异表达基因80.1%未见报道。结论综合运用交集补集分析、t检验、秩和检验对基因谱数据进行挖掘的策略,可为寻找大肠癌分子标记物和从整体上探讨大肠癌发生的分子机制奠定基础。
Objective To screen differentially expressed genes in human colorectal carcinoma and corresponding normal colorectal mucosa. Methods Affymetrix HG-U133 oligonucleotide microarray (representing 32,664 human whole genomes known to date, including 19,308 known human genes and 12,956 EST clusters) was used to detect the gene expression of 9 cases of colorectal carcinoma and corresponding normal colorectal mucosa Spectrum, and real-time fluorescence quantitative PCR technology to verify the results of the gene chip; comprehensive use of intersection complement, rank sum test and t test to compare the two sets of expression data. Results There were 3 125 differentially expressed genes and ESTs in colorectal cancer tissues and normal colorectal mucosa tissues (including 974 ESTs of up-regulated genes and 2115 ESTs of down-regulated genes), 245 ESTs expressed in colorectal cancer tissues and corresponding normal mucosa tissues, There were 162 ESTs expressed in normal mucosa and 17 ESTs of differentially expressed genes in colorectal cancer. 80.1% of the differentially expressed genes in colorectal cancer screened in this study have not been reported. Conclusion The strategy of using the complement complement analysis, the t-test and the rank sum test to excavate the gene profiling data can lay a foundation for finding the molecular markers of colorectal cancer and exploring the molecular mechanism of colorectal cancer as a whole.