基因表达谱芯片发现36条喉鳞癌相关基因

目的：研究探讨喉鳞癌发生、发展中相关基因群的表达和初步功能。方法：按微矩阵排列的4096种全长基因PCR产物制成BioDoor4096型微矩阵表达谱芯片；采用条件优化的一步法抽提喉鳞癌及正常组织总RNA，用Qiagen公司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，芯片杂交和严格洗片后，用SanArray3000荧光扫描仪扫描芯片荧光信号图像，利用计算机分析肿瘤及正常组织中差异表达的基因。对所获得的基因进行分子生物信息学分析。结果：在4096种基因中，喉鳞状细胞癌与正常组织间存在差异表达的基因。在所检测的4对临床标本中，发现有差异表达的基因36条（0．88％）。生物信息学分析显示，该36条差异表达基因与肿瘤的发病机制可能存在相关性。结论：喉鳞状细胞癌的发生、发展中存在多因素表达调控的改变，对于相关基因群的研究有助于认识肿瘤发病机制。

Study of differentially expressed genes in laryngeal squamous cell carcinoma by cDNA microarray

Objective: To screen for the differentially expressed genes in laryngeal squamous cell carcinoma and normal laryngeal tissue using cDNA microarray. Methods: The PCR products of 4 096 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 were purified to mRNAs by Oligotex. Both the mRNAs from the laryngeal squamous cell carcinoma and normal 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 high stringent washing, the cDNA microarray was scanned for the fluorescent signals and showed the differences between 2 tissues. Results: Among the 4 096 target genes, there were 36(0.88%) genes whose expression levels differed between the carcinoma and normal tissues in all 4 cases. Bioinformatical analysis of those genes had been performed. Conclusion: DNA microarray technology is an effective technique in screening for differentially expressed genes between 2 different kinds of tissue. Further analysis of the obtained genes will help to understand the molecular mechanism of malignant carcinoma.