基于胶质母细胞瘤干细胞的高通量抗肿瘤药物筛选技术的建立和应用

项目名称： 基于胶质母细胞瘤干细胞的高通量抗肿瘤药物筛选技术的建立和应用

项目编号： No.81472862

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： 赵旭东

作者单位： 中国科学院昆明动物研究所

项目金额： 80万元

中文摘要： 肿瘤干细胞的存在近年来获得了大量的实验证据支持，是肿瘤耐药、复发和转移的主要原因，理应是肿瘤治疗的主要靶细胞。目前的抗肿瘤药物都是用肿瘤细胞系筛选出来的，而抗肿瘤干细胞的药物将具有更好的肿瘤治疗效果，但目前缺乏针对肿瘤干细胞的药物，其重要原因是没有基于肿瘤干细胞的良好的高通量筛选技术。与其他肿瘤干细胞相比，胶质母细胞瘤干细胞可长期体外、贴壁培养，并且传代过程中遗传学、生物学特性等不易改变，具有用于高通量筛选的潜力。本项目拟遴选有代表性基因变异的胶质母细胞瘤干细胞株，将报告基因EGFP定点整合在胶质母细胞瘤干细胞分子标记Nestin启动子下游，利用细胞高内涵筛选系统，建立可同时筛选细胞毒性和分化诱导药物的技术；利用这一技术将对天然产物库及其他化合物库进行抗肿瘤药物筛选，获得针对肿瘤干细胞的先导化合物并对部分化合物抗肿瘤效果和机制进一步研究。

中文关键词： 胶质母细胞瘤干细胞；高通量；药物筛选

英文摘要： Theory of tumor stem cell is getting more and more experimental supports and tumor stem cell is widely accepted as a main cause of tumor chemoresistance, radioresistance, tumor migration and recurrence. The tumor stem cell is the ideal target of therapy. Theoretically drugs against tumor stem cell should have better anti-tumor effect. However current anti-tumor drugs were screened out by using cancer cell lines. It is necessary to use tumor stem cell for anti-tumor drug screening. However there are not many efforts on screening with tumor stem cells. The main reason is lacking a good high-throughput screening system by using tumor stem cell. Glioblastoma stem cell is one of a few cancer stem cells that can be grown and passaged in vitro for long time both as neurosphere and adherent culture without obvious change in genetic and gene expression profile, which are the prerequisite for high-throughput screening. In this study，1) we will establish a glioblastoma stem cell-based high-throughput screening system by homologous integration of reporter gene EGFP into downstream of stem cell marker gene Nestin promoter. With this system,both cytotoxic and differentiation-inducing compounds can be screened simultaneously. The single cell information from high content screening technology can discriminate cytotoxicity and differentiation induced by compounds；2) Compound library, specifically natural compound and conventional drugs etc., will be screened; 3) In vivo anti-tumor effect and mechanism of selected candidate from screening will be elucidated. The screening system and compound candidate will undoubtedly promote development of anti-tumor stem cell drugs.

英文关键词： glioblastoma stem cell;high-throughput;compound screening