胆囊癌细胞恶性优势克隆的转移潜能分析

项目名称： 胆囊癌细胞恶性优势克隆的转移潜能分析

项目编号： No.30872502

项目类型： 面上项目

立项/批准年度： 2009

项目学科： 轻工业、手工业

项目作者： 刘颖斌

作者单位： 浙江大学

项目金额： 32万元

中文摘要： 本课题组运用肿瘤表达谱基因芯片技术对胆囊癌细胞GBC-SD进行了基因表达谱的检测。通过生物信息学的分析，得到了在GBC-SD中高表达和低表达的分子群。通过我们的比较和分析后发现， maspin在GBC-SD中表达显著性降低。通过建立基于细胞水平的药物筛选平台，我们首次报道了Wogonin可以增加maspin基因表达水平并进而抑制胆囊癌细胞的运动能力和侵袭转移能力。我们还发现，cirsimaritin和matrine对胆囊癌细胞有显著地诱导凋亡的作用，并阐明了介导这一功能的分子机制。另外，我们还通过对芯片结果的分析，发现NLK和ZFX在GBC-SD 中高表达。通过RNAi 的实验方法在GBC-SD细胞内分别沉默NLK 和ZFX 这两个分子后，Transwell 实验证实，GBC-SD的迁移能力显著性地降低，提示NLK和ZFX可能是导致胆囊癌细胞发生侵袭转移的关键分子。最后通过建立胆囊癌血行转移模型和淋巴转移模型，为研究胆囊癌的转移机制提供了更为有效的实验工具。通过以上这些研究结果，加深了我们对于胆囊癌转移过程的认识，为我们针对胆囊癌的侵袭转移过程设计特异性的靶向药物提供了有效的分子靶点。

中文关键词： 胆囊癌；基因；凋亡；

英文摘要： The gene expression profile of gallbladder carcinoma cell line GBC-SD was accessed by using microarray of tumor genes. All differentially expressed genes between GBC-SD and control group were evaluated with bio-informatics. We found that the expression level of maspin decreased dramatically in GBC-SD cells as compared to that of control. Based on drug screening platform of cellular level, we reported for the first time that the expression level of maspin increased significantly after stimulation of wogonin which can in turn inhibit the migration and invasion of GBC-SD cells. Furthermore, cirsimaritin and matrine were also discovered to play an important role in apoptosis based on the same drug screening platform. In the following study, the molecular mechanisms by which cirsimaritin and matrine exerted their functions on apoptosis were explored and evaluated. Otherwise, NLK and ZFX were found to be highly expressed in GBC-SD cell which implied a key role in GBC-SD. We found that the metastasis of GBC-SD cells with gene silencing of NLK or ZFX by lentivirus-mediated RNAi was impaired dramatically, suggesting that these two molecules may have important roles in cancer metastasis. Two experimental models, spleen-liver metastasis model and footpad-inguinal lymph node metastasis model, were established in order to provide us a powerful tool to investigate the mechanism of metastasis in vivo and widen our knowledge about the carcinoma progression of gallbladder cancer. Overall, all our results suggested the important signaling pathways in metastasis progression and provided us potential therapeutic markers for gallbladder carcinoma.

英文关键词： gallbladder carcinoma;gene;apoptosis