基因芯片在胰腺癌相关基因筛选中的初步研究

目的探讨基因表达谱芯片技术在筛查胰腺癌相关基因群表达中的作用。方法按微矩阵排列的4096种全长基因PCR产物制成Biostar H-40s型微矩阵表达谱芯片;采用一步法抽提胰腺癌及正常胰腺组织总RNA,用Oligotex mRNA离心柱分离纯化两种组织的mRNA;经逆转录合成荧光分子(Cy3/Cy5)掺入的cDNA一链制备表达谱探针,芯片杂交和洗片后,用荧光扫描仪扫描芯片荧光信号图像,分析胰腺癌及正常胰腺组织中差异表达的基因。结果在4096种基因中,胰腺癌组织与正常胰腺组织间有949条(23.2%)存在差异表达的基因,其中我们确定了9条上调和8条下调的癌基因和抑癌基因可能与胰腺癌的发病机制存在相关性。结论基因表达谱芯片技术可以筛选出胰腺癌表达异常的相关基因群,对其进一步研究有助于认识胰腺癌的发病机制。

Study of Differentially Expressed Genes in Pancreatic Cancer by cDNA Microarray

Objective To screen for the differentially expressed genes in pancreatic carcinoma and normal pancreatic tissue using cDNA microarray. Methods The PCR products of 4096 genes were spotted on a chemical-material-coated-glass plates in array. The DNAs were then fixed on the glass plate by a serial of treatments. The total RNAs were isolated from the tissues, and then mRNAs were purified to by Oligotex. Both mRNAs from pancreatic carcinoma and normal pancreatic tissue were reversely transcribed to cDNAs with the incorporations of fluorescent dUTP, for preparing the hybridization probes. The mixed probes were then hybridized to the cDNA microarray. After washing, the cDNA microarray was scanned. Results Among the 4 096 target genes, there were 949( 23.2% ) genes whose expression levels differed between the carcinoma and normal tissues. We identified 9 upregulated and 8 downregulated genes belonging to oncogenes and anti-oncogenes. Conclusion cDNA microarray technology is a powerful method in screening for differentially expressed genes. Further analysis of the obtained genes will help to understand the molecular mechanism of pancreatic carcinoma.