基因芯片技术筛选大肠癌肝转移差异表达基因

A choosing of the differentially expressed genes in colorectal cancer liver metastasis with gene microarray

目的:利用基因芯片技术研究大肠癌肝转移基因表达的改变及其相关基因。方法:收集手术切除5例大肠癌合并肝脏转移患者的原发和转移病灶的新鲜组织标本,提取总RNA,合成荧光标记的cRNA与Agilent human 1A oligo芯片杂交。挑选10个差异表达基因,分别设计引物,用实时RT-PCR法定量测定21例术中收集的新鲜原发和转移标本中各基因的表达水平,并比较和分析其表达差异。结果:以表达差异≥2.0或≤0.5倍为限,在5对标本中,共同上调110个基因,共同下调96个基因。挑选10个差异表达基因的实时RT-PCR结果与芯片结果完全相符。结论:大肠癌肝转移涉及众多基因表达的改变,应用基因微矩阵有助于发现基因变化的规律及其分子机理的深入研究。

Objevtive：To explore the gene expressive changes and related genes associated with colorectal cancer and hepatic metastases with gene microarray. Methods：The fresh specimens of primary cancer lesion and hepatic metastasis were collected in 5 patients suffered from colorectal cancer with liver metastasis during operation. After the total RNA extraction,the synthesized biotinylated cRNAs were hybridized to Agilent human 1A oligo microarray. After the design and synthesis of the primers, 10 of the differentially expressed genes were then chosen at random to confirm the array results using the SYBR Green Real-time RT-PCR method in 21 fresh specimens collected from primary cancer lesion and hepatic metastasis.The data analysis depended on the ABI Prism 7500 system. Results：Using an extrance limit of≥ 2.0 or ≤ 0.5, there were 110 up-regulated and 96 down-regulated genes identified by mieroarray which were commonly differentially expressed among the 5 cases. The array results were further confirmed by the real-time RT-PCR. Conclusion：Many genes were involved in the colorectal cancer with liver metastases. It could be helpful to find the regularity of the gene expression by gene microarray and to study the molecular mechanism in the colorectal cancer with liver metastases.