基于功能激酶组学的新型EMT调节分子的筛选验证及调节机制研究

项目名称： 基于功能激酶组学的新型EMT调节分子的筛选验证及调节机制研究

项目编号： No.81472687

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 医药、卫生

项目作者： 李琳娜

作者单位： 中国人民解放军军事科学院军事医学研究院

项目金额： 64万元

中文摘要： EMT与肿瘤转移密切相关。已知的EMT调节分子中，可作为药物靶标者为数甚少。激酶是理想的药物作用靶点，我们前期利用EMT标记物Vimentin的报告载体，从激酶组cDNA文库中筛选出新型EMT调节分子，之后构建稳定表达这些激酶的细胞株，验证其表型和功能，得到有效调节EMT激酶。通过NCBI GO数据库分析有效激酶的已知功能，发现大多与细胞周期、代谢、自噬和组蛋白修饰相关。据此，我们提出能源诉求驱动肿瘤细胞发生EMT的作用机制：当肿瘤细胞感知能源匮乏时，会通过增强糖酵解、增强自噬和使细胞阻滞于G0/G1期，为其合成代谢提供更多原料、中间产物和创造最佳时机。同时，组蛋白修饰酶作为传感器，将代谢产物的异常变化通过表观遗传重塑转化为基因表达的变化，使肿瘤细胞发生EMT，转移到能源丰富的器官。本课题将为全面理解肿瘤细胞EMT提供全新的视角和可靠的实验依据，为新型EMT调节激酶的药物研发奠定基础。

中文关键词： 上皮细胞间充质转化；激酶组；肿瘤转移；药物靶标；细胞代谢

英文摘要： The epithelial-mesenchymal transition (EMT) is closely associated with cancer metastasis. However, few EMT regulators can be used as cancer drug targets. Since kinases have been recognized as ideal drug targets, we carried out human kinome cDNA screens that surveyed 500 human kinases to identify novel EMT regulators. In this screen, we used vimentin promoter luciferase assay to identify potential EMT regulators, and then validated their EMT-inducing function by establishing stable cell lines. Furthermore, NCBI GO analysis uncovered that most effective EMT-inducing kinases were associated cell cycle regulation, cell metabolism, autophagy, or histione modification. According these results, we came up with a hypothesis explaining EMT of cancer cells: when cancer cells feel hungry, they will fulfill their exuberant growth requirement by enhancing glycolysis, autophagy or promoting G0/G1 arrest, through which they get raw material, intermediate products, and best opportunity for anabolic metabolism. At the same time, the histone, the sensor of abnormal metabloites, could transfer the changes of cell metabolism into changes of some genes expression through reprogramming cell epigenetic modification. Therefore, cancer cells undergo EMT and spread into other organs with enough energy to furfill their strong requirement for energy. Through identifying novel EMT regulators from human kinome cDNA screens and uncovering their regulation mechanism, this study will open a new window to see EMT and provide new potential targets for anticancer drug development.

英文关键词： EMT;Kinome;Metastasis;Drug target;Metabolism