采用基因表达谱芯片筛选肺癌细胞系差异表达基因的初步探讨

Detection of Differentially Expressed Genes in the Hunman Lung Carcinoma Cell Line with Different Metastatic Phenotype

目的:探讨基因芯片技术分析高低不同转移能力肺癌细胞系的差异表达基因。方法:采用基因芯片技术检测具有高低不同转移能力的肺癌细胞系BE-1和LH-7的差异表达基因。抽提细胞mRNA,利用荧光标记dUTP逆转录制备cDNA探针,与人肺癌表达谱基因芯片杂交,以ScanArray4000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的比值。结果:在所检测的人高低肺癌转移细胞系BE-1和LH-7中,20个基因有表达差异,其中高表达14条,低表达6条。结论:基因芯片技术为筛选人类肺癌转移基因提供了有效方法。

Objective: The analysis on differential gene expression profiles of human lung carcinoma cell lines with different metstatic phenotype. Methods: To prepare the probes,mRNA from cells were isolated and purified, then reversely transcribed to cDNA with the incorporation of fluorecent_labeled dUTP. The probes were hybridized with a cDNA microarray. The chip were scanned by ScanArray 4000 laser scanner on two wavelengths. The acquired image was analyzed by Gene Pix Pro 3.0 software. The intensity of each spot on the two wavelengths presented the quantity of Cy3-dUTP and Cy5-dUTP, with Cy5 to Cy3 ratio computed on each. Results: 20genes exhibited differential expressions in 2 lung carcinoma cell lines. 6genes were down-regulated, 14 gene were up-regulated. Conclusion: cDNA microarry provides an promising approach to specific gene expressions of the presence of metastasis in human lung carcinoma.