FGFR4/CMC-HPLC/MS筛选白芥子促肿瘤细胞凋亡活性成分研究

项目名称： FGFR4/CMC-HPLC/MS筛选白芥子促肿瘤细胞凋亡活性成分研究

项目编号： No.81202495

项目类型： 青年科学基金项目

立项/批准年度： 2013

项目学科： 药物学、药理学

项目作者： 张涛

作者单位： 西安交通大学

项目金额： 23万元

中文摘要： 利用以受体靶标为核心的药物直接筛选技术，从药用植物中寻找高效、低毒抗肿瘤活性成分是药物研究的热点内容之一。抑制酪氨酸激酶受体家族成员FGFR4在癌细胞中的表达可促进肿瘤细胞凋亡，提高肿瘤细胞药物敏感性。本项目在前期工作的基础上，利用细胞生物学、色谱科学、药物分析等技术，以FGFR4为研究对象，构建HEK293-FGFR4高表达细胞系，建立高表达FGFR4/CMC-HPLC/MS筛选模型，筛选、鉴定中药白芥子中可与FGFR4相互作用的活性成分；通过MTT、cell cycle、apoptosis analysis、放射性标记化合物标记分析、QT-PCR、western blot等方法，分析活性成分促肿瘤细胞凋亡的作用及分子机制，并验证其抗肿瘤活性。本项目不仅为阐明FGFR4作为抗肿瘤增敏药物筛选靶标提供实验依据，同时为筛选抗肿瘤天然活性成分提供新的研究模型，为抗肿瘤药物筛选开拓思路。

中文关键词： 成纤维细胞生长因子；细胞膜色谱；白芥子；活性成分筛选；凋亡

英文摘要： It is a hotspot for tumor treatment research to looking for safe and effective anti-tumor components from traditional Chinese herbal medicine using receptor-based parmaceutical screening techology. Trysoine kinase receptor have been proved to be an excellent drug screening target for cancer therapy. Inhibiton of fibroblast growth factor receptor 4 (FGFR4) can promot cancer cell apoptosis and sensitive chemo-drug. In present study, we firstly establish overexpressing HEK293-FGFR4 cells by stabilized transfection method. Based on techniques of pharmaceutical analysis and cell membrane chromatography, HEK293-FGFR4 cells are applied in FGFR4/CMC-HPLC/MS model to screening and identify active components from Sinapi alba L. Then, we could elucidate the effect and mechanism of screened active components on cancer cell apoptosis and increasing drug sensitivity via in vitro experiment MTT, cell cycle, apoptosis analysis, QT-PCR, western blot. The research can biuld a new model for screening natural anti-tumor components. we can also elucidate that FGFR4 is the drug screening target for chemosensitivity. This study is benficial to extending our mind on screening anti-tumor drug.

英文关键词： FGFR4；cell membrane chromatography；Brassica alba L.；screening active；apoptosis