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

采用寡核苷酸扩增法构建了口腔鳞状细胞癌（SCC）和涎腺肿瘤（SGT）的cDNA文库。测定了cDNA克隆的5 '端核苷酸序列。排除重叠克隆，选择2，201个独立克隆并用于微阵列生产。与公开的核苷酸序列数据库相比，我们的cDNA克隆的61%是全长的。使用该微阵列系统分析了从鳞状细胞癌（SCC）和腺样囊性癌（ACC）获得的mRNA。基因芯片检测到许多基因表达上调，在肿瘤组织中的表达高于正常组织。结果表明，在SCC中表达差异的全长克隆有IL-1 β、Centaurin γ 2、Adenyl ...更多信息 γ 1环化酶相关蛋白、Rh C型糖蛋白、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、假想蛋白FLJ 20150、富含脯氨酸的小蛋白、Carboxylink蛋白、G蛋白α 15、白细胞介素受体拮抗剂、转铁蛋白等，而在SGT中差异表达的基因为补体衰变加速因子（CD 55）、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、人波形蛋白基因、免疫球蛋白重链恒定区μ、T细胞受体β链、脱氢酶/还原酶成员2、18 S rRNA基因等。通过对临床标本的功能分析和免疫组化染色，发现了几种重要的生物学标志物。Clorf 10基因是一种肿瘤抑制基因，可阻止细胞周期，Rab la是致癌作用预测因子的候选基因，maspin是ACC特异性分子标记物，stathmin是恶性肿瘤特异性分子标记物。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