Search for oral cancer specific genes and molecular cloning using in-house cDNA microaray

以口腔鳞状细胞癌（SCC）和唾液腺肿瘤（SGT）的手术标本为样本，采用寡聚分析法构建cDNA文库。测定cDNA克隆的5′端核苷酸序列。排除重叠克隆，选择2201个独立克隆用于微阵列生产。与公共核苷酸序列数据库相比，61%的cDNA克隆是全长的。利用该微阵列系统分析了从鳞状细胞癌（SCC）和腺样囊性癌（ACC）中获得的mrna。微阵列检测到许多基因表达上调，这些基因在正常组织中表达两倍或更多。通过微阵列分析，我们可以提取出全长的完整基因，并直接使用全长克隆来检测基因的功能。SCC中具有差异上调/下调表达的代表性全长克隆有白细胞介素1 β、半人马座γ 2、腺苷…More酰基环化酶相关蛋白、Rh型C糖蛋白、精胺合成酶、Copine 1、细胞角蛋白13、配对样同源结构域转录因子1、假设蛋白flj2015、富含小脯氨酸蛋白、软骨连接蛋白、g蛋白α 15、白细胞介素受体拮抗剂、转谷氨酰胺等。而在sgs中差异表达的基因有补体衰变加速因子（CD55）、诱导信号通路蛋白2、胰岛素样生长因子结合蛋白3、fos样抗原1、人vimentin基因、免疫球蛋白重常数mu、T细胞受体β链、脱氢酶/还原酶成员2、18S rRNA基因等。通过edcsone启动子系统和临床样品的免疫组织化学染色分析，发现了一些重要的生物标志物。clorf10基因是一种肿瘤抑制因子，可以阻止细胞周期，Rab - la是一种候选的癌症发生预测因子，maspin是ACC特异性的分子标记物，而stathmin是恶性肿瘤特异性的分子标记物。少

cDNA library was constructed from surgical specimens of oral squamous cell carcinoma(SCC) and salivary gland tumors(SGT) using an oligo-eapping method. The 5 '-end nucleotide sequences of the cDNA clones were determined. Overlapping clones were excluded, and 2,201 independent clones were selected and used for microarray production. Compared to the public nucleotide sequence database, 61% of our cDNA clones were full length. mRNAs obtained from squamous cell carcinomas(SCC) and adenoid cystic carcinomas(ACC) were analyzed using this microarray system. Microarray detected up-regulated expression of many genes, which expressed twice or more in tumors man in normal tissue. We could pick up the full length whole gene, which was focused on by microarray analysis, and were directly and immediately able to examine the gene function using the full length clone.Representative full length clones with differential up-/down-regulated expression in SCC were Interleukin 1 β, Centaurin gamma 2, Adenyl … More yl cyclase-associated protein, Rh type C glycoprotein, Spermine syntase, Copine 1, Cytokeratin 13, Paired-like homeodomain transcription factor 1, Hypothetical protein FLJ20150, Small proline-rich protein, Cartilage link protein, G-protein alpha 15, Interleukin receptor antagonist, Transglutamin and so on, whereas the genes differentially expressed in SGTs were decay accelerating factor for complement (CD55), Inducible signaling pathway protein 2, Insulin-like growth factor binding protein 3, Fos-like antigen 1,Human vimentin gene, Immunogloblin heavy constant mu, T cell receptor beta chain, Dehydrogenase/reductase member 2,18S rRNA gene and et. The fgene function -nalysis by edycsone-promoter system and immunohistochemical staining of clinical samples, revealed several important bio-markers. Clorf 10 gene is a tumor suppressor, which stops cell cycle, Rab la is a candidate for a predictor of carcinogenesis, maspin is a molecular marker specific to ACC, and stathmin is a molecular marker specific to malignancy. Less