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目的 :应用基因微矩阵方法筛选喉鳞癌相关基因。方法 :按一步法抽提喉鳞癌和对照喉正常组织的总 RNA并纯化m RNA;将 40 96种人类基因 PCR产物用 Cartesian Pixsys75 0 0点样仪按微矩阵排列点样于化学涂层的载玻片上 ,制成基因芯片 ;将等量的对照组织和喉鳞癌组织 m RNA分别逆转录合成荧光分子掺入的 c DNA一链做探针 ,混合后杂交上述基因芯片。经严格洗片后用 Scan Array30 0 0扫描仪扫描芯片荧光信号图像 ,计算机分析后比较两种组织中差异表达的基因。结果 :在 40 96种基因中 ,喉鳞状细胞癌与喉正常组织间存在差异表达的基因。所检测的 4例临床标本中 ,2例共有的差异表达基因2 11～ 35 6条 (5 .15 %～ 8.87% ) ,3例共有的差异表达基因 36条 (0 .88% )。结论 :基因微矩阵芯片在筛选喉肿瘤发生相关基因的改变具有快速、高通量、高敏度等特点。
Objective: To screen genes associated with laryngeal squamous cell carcinoma using gene microarray. Methods: One-step extraction of total RNA from laryngeal squamous cell carcinoma and control laryngeal normal tissue and purification of m RNA; 40 96 human gene PCR products were arrayed on a chemical matrix using a Cartesian Pixsys 7500 spotter. On the glass slide, a gene chip was made; an equal amount of the control tissue and m RNA of laryngeal squamous cell carcinoma were respectively reversely transcribed into a fluorescent DNA-incorporated c-DNA strand as a probe, and the above gene chip was hybridized after mixing. Strictly washed films were scanned with a Scan Array 3000 scanner and the computer-analyzed genes were differentially expressed in both tissues. RESULTS: Among 40 96 genes, there were differentially expressed genes between laryngeal squamous cell carcinoma and normal laryngeal tissue. Of the 4 clinical specimens tested, 2 were differentially expressed genes from 11 to 356 (5.15% to 8.87%), and 3 differentially expressed genes were 36 (0.88%). Conclusion: The gene microarray chip has the characteristics of rapid, high-throughput and high-sensitivity in the screening of genes involved in laryngeal tumorigenesis.