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

使用内部 cDNA 微阵列对腺样囊性癌细胞进行基因分析

• 批准号: 15592096
• 负责人: YOKOE Hidetaka
• 金额: $ 2.24万
• 依托单位: Chiba University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15592096/
• 关键词: adenoid cystic carcinoma; microarray analysis; salivary gland tumor; maspin; stathmin; 線様嚢胞癌; マイクロアレイ; cDNA; 遺伝子発現プロファイリング; 唾液腺腫瘍

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.

我们创建了一个内部cDNA微阵列，其中包含源自口腔鳞状细胞癌（SCC）cDNA库和778个cDNA克隆的1,423个cDNA克隆，这些cDNA cDNA cDNA cDNA cDNA cDNA库。使用此微阵列系统分析了从起源于SGT的腺样性囊性癌（ACC）细胞获得的mRNA。此外，还使用附着38,000多个基因的Affymetrix Genechip^<tm>进行了更多的微阵列测定，以检查更多的基因。微阵列检测到许多基因的上调表达，其在ACC中的表达比正常的唾液腺组织表达了两倍或更高。检测到的代表性上调的基因是FAS诱导的凋亡，肿瘤坏死因子超级肝脏，MASPIN，ANNEXIN A8，成纤维细胞生长因子受体1，胰岛素样生长因子2，Fibrin2，纤维蛋白2，淀粉样蛋白β4，蛋白酶4，4，胶原蛋白，胶原蛋白和laminin和laminin。另一方面，代表性下调的基因由WT1，BCL-2，非纤维蛋白，脂肪肿瘤抑制剂同源物，Notch，S100钙结合蛋白A1，趋化因子，肿瘤坏死因子超家族，Stathmin，cd44和lysozyme组成。在上调的基因中，选择了Maspin和Stathmin进行进一步的分析，包括蛋白质印迹和免疫组织化学染色。与口服SSCC相比，我们发现Maspin在中士中的表达更高，这表明Maspin是特定于ACC的分子标记。有趣的是，ACC和SCC的蛋白质mRNA和蛋白质表达水平均明显更高，表明斯塔明蛋白是特异性的分子标记。