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目的 应用cDNA芯片技术进行肝外胆管癌相关基因表达谱差异分析 ,筛选胆管癌相关基因。方法 按一步法分别抽提 6例胆管癌和正常人胆管黏膜总RNA、纯化 ,逆转录合成掺入荧光分子的cDNA链探针 ,与 10 68条人PCR微矩阵芯片杂交 ,扫描芯片荧光信号图像 ,计算机分析比较二种组织基因表达谱差异。结果 胆管癌与正常胆管黏膜的基因表达谱分析 ,发现有 194条基因表达差异 ,6组标本 47条与肿瘤相关的基因一致向上或向下表达 ,2 3条表达上调 ,2 4条表达下调。结论 基因芯片能快速筛选胆管癌相关基因 ,分析这些差异表达的基因能够阐明胆管癌复杂的生物学特性与基因表达之间的内在联系 ,并识别肿瘤的标记物。
Objective To analyze the gene expression profiles of extrahepatic cholangiocarcinoma by cDNA microarray and screen the genes related to cholangiocarcinoma. Methods Total RNA was extracted from bile duct mucosa of 6 cases of cholangiocarcinoma and normal human by one-step method. The cDNA strand probe that incorporates fluorescent molecules was purified and reverse transcribed. The PCR products were hybridized with 10 68 PCR microarray chips, , Computer analysis and comparison of two kinds of tissue gene expression differences. Results There were 194 gene expression profiles in cholangiocarcinoma and normal bile duct mucosa. 47 of the 6 specimens were up-and down-regulated in association with the tumor-associated genes, 23 up-regulated and 24 down-regulated. Conclusion Gene chips can rapidly screen cholangiocarcinoma related genes and analyze these differentially expressed genes can clarify the intrinsic relationship between the complex biological characteristics of cholangiocarcinoma and gene expression and identify tumor markers.