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目的 利用基因芯片技术分析伴与不伴淋巴结转移胰腺癌组织中的差异表达基因。方法 将 40 0 0个人类全长基因的PCR产物点样于特殊处理后的玻片上制备基因芯片。用逆转录的方法 ,分别将两种荧光Cy3 dUTP和Cy5 dUTP标记正常胰腺组织和胰腺癌组织mRNA ,以制备cDNA探针 ,并与芯片杂交。以ScanArray 30 0 0荧光扫描仪扫描芯片上两种荧光信号 ,获得的荧光信号图像用计算机分析。每点上两种荧光信号的强度分别代表Cy3 dUTP和Cy5 dUTP的量。最后 ,计算每点的Cy5和Cy3比值。结果 在所检测的 2例伴淋巴结转移和 2例不伴淋巴结转移的胰腺癌标本中 ,5 6个基因 (其中包括 2 4个已知基因 )有表达差异。结论 基因芯片技术为阐明人类胰腺癌伴淋巴结转移的特异性基因表达提供了有效方法。
Objective To analyze differentially expressed genes in pancreatic cancer with or without lymph node metastasis by using gene chip technique. Methods PCR products of 400 human full-length genes were spotted onto specially treated slides to prepare gene chips. Reverse transcription method was used to label two fluorescent Cy3 dUTP and Cy5 dUTP respectively for mRNA of pancreatic and pancreatic tissues to prepare cDNA probe and hybridize with the chip. The two fluorescence signals on the chip were scanned with a ScanArray 30 0 0 fluorescence scanner and the fluorescence signal images obtained were analyzed by a computer. The intensities of the two fluorescent signals at each point represent the amounts of Cy3 dUTP and Cy5 dUTP, respectively. Finally, calculate the Cy5 and Cy3 ratio for each point. Results In the two cases of pancreatic cancer with lymph node metastasis and two cases without lymph node metastasis, 56 genes (including 24 known genes) were found to be differentially expressed. Conclusion Gene chip technology provides an effective method to elucidate the specific gene expression of human pancreatic cancer with lymph node metastasis.