Gene profiling in adenoid cystic carcinoma cells using in-house cDNA microaray

我们建立了一个内部的cDNA微阵列，包含1,423个来自口腔鳞状细胞癌（SCC）cDNA文库的cDNA克隆和778个来自唾液腺肿瘤（SGT）cDNA文库的cDNA克隆。使用该微阵列系统分析从源自SGT的腺样囊性癌（ACC）细胞获得的mRNA。此外，<TM>为了检测更多的基因，使用附着了超过38，000个基因的Affyssin GeneChip ®进行了更多的微阵列测定。基因芯片检测到多个基因表达上调，其中ACC中的表达是正常涎腺组织的两倍或更多。检测到的代表性上调基因是Fas诱导的细胞凋亡调节因子、肿瘤坏死因子超家族、maspin、膜联蛋白A8、成纤维细胞生长因子受体1、胰岛素样生长因子2、纤维蛋白2、淀粉样蛋白β 4、胶原蛋白和层粘连蛋白。另一方面，代表性下调基因包括WT 1、BCL-2、Inhibitin、FAT肿瘤抑制基因同源物、Notch、S100钙结合蛋白A1、趋化因子、肿瘤坏死因子超家族、stathmin、CD 44和溶菌酶。在上调的基因中，选择maspin和stathmin用于进一步分析，包括Western印迹和免疫组织化学染色。我们发现，与口腔sSCC相比，SGT中maspin的表达更高，表明maspin是ACC特异性的分子标记物。有趣的是，stathmin mRNA和蛋白表达水平在ACC和SCC中均显著更高，表明stathmin是恶性肿瘤特异性的分子标记物。

We created an in-house cDNA microarray containing 1,423 cDNA clones derived from an oral squamous cell carcinoma(SCC) cDNA library and 778 cDNA clones derived from a salivary gland tumor(SGT) cDNA library. mRNAs obtained from adenoid cystic carcinoma(ACC) cells that originated from SGT were analyzed using this microarray system. Additionally, further more microarray assay was performed with Affymetrix GeneChip^<TM>, which attached more than 38,000 genes, in order to examine more genes. Microarray detected up-regulated expression of many genes, which expressed twice or more in ACC than in normal salivary gland tissue. Representative up-regulated genes detected were regulator of Fas-induced apoptosis, tumor necrosis factor superfamily, maspin, annexin A8,fibroblast growth factor receptor 1,insulin-like growth factor 2,fibrin2,amyloid beta 4,collagen, and laminin. On the other hand, representative down-regulated genes consisted of WT1,BCL-2,Inhibrin, FAT tumor suppressor homolog, Notch, S100 calcium binding protein A1,chemokine, tumor necrosis factor superfamily, stathmin, CD44,and lysozyme. Of the up-regulated genes, maspin and stathmin were selected for further analyses including Western blotting and immmunohistochemical staining. We found higher expression of maspin in SGT compared with oral sSCC, indicating that maspin is a molecular marker specific to ACC. Interestingly, stathmin mRNA and protein expression levels were significantly higher in both ACC and SCC, indicating that stathmin is a molecular marker specific to malignancy.