基于RAC1/NADPH信号通路对蒽醌化合物GXHSWAQ-3增强鼻咽癌辐射敏感性的时空调控机制

项目名称： 基于RAC1/NADPH信号通路对蒽醌化合物GXHSWAQ-3增强鼻咽癌辐射敏感性的时空调控机制

项目编号： No.81460561

项目类型： 地区科学基金项目

立项/批准年度： 2015

项目学科： 医药、卫生

项目作者： 侯华新

作者单位： 广西医科大学

项目金额： 51万元

中文摘要： 国内外研究及本课题组发现，RAC1/NADPH通路与辐射产生活性氧致细胞氧化性损伤密切相关。但RAC1/NADPH通路的调控与辐射敏感关系的研究甚少。本课题以具有放射增敏活性GXHSWAQ-3和不同分化能力的鼻咽癌细胞为对象，采用蛋白印迹、免疫共沉淀、流式细胞术检测GXHSWAQ-3联合射线作用细胞后， RAC1/NADPH信号通路上各亚基的表达和细胞存活情况。FRET结合激光共聚焦显微镜动态监测GXHSWAQ-3作用后不同时点RAC1与NADPH酶亚基结合、转位、分布及蛋白复合物形成的情况，明确GXHSWAQ-3调控不同分化鼻咽癌细胞RAC1与NADPH酶亚基相互作用的时空关系。构建RAC1低表达和过表达的细胞并接种于裸鼠，明确体内RAC1/NADPH信号通路调控与鼻咽癌辐射敏感的关系，建立分子成像研究小分子药物与调控靶蛋白信号通路的新方法，开创一个筛选鼻咽癌放射增敏剂全新的研究平台。

中文关键词： RAC1/NADPH；信号通路；蒽醌化合物；放射敏感性；鼻咽癌；时空调控

英文摘要： The domestic and foreign research and our studies supported RAC1/ NADPH pathway was involved in cell oxidative damage by reactive oxygen species(ROS) during radiation process. But the study on radiosensitizer regulation of the RAC1/ NADPH pathway is very little. GXHSWAQ-3 , a radiosensitizing activity compound ,was synthesised by our preliminary work. To explore the potential mechanisms involved of GXHSWAQ-3 enhanced different differentiation nasopharyngeal carcinoma cells on X-ray sensitive，the expression of RAC1 and the subunites of NADPH in the RAC1/NADPH pathway and the cell viability are detected using Western blotting, co-immunoprecipitation , flow cytometry . FRET combine with confocal microscopy will be used to dynamic monitoring the distribution of RAC1 activation in cells location, distribution and NADPH oxidase complex formation in different differentiation nasopharyngeal carcinoma cells, to reveal the key role that GXHSWAQ-3 targeting RAC1/NADPH pathway in the regulation of RAC1 interaction with subunites of NADPH oxidase. Over-expressing RAC1 and RAC1 RNAi cell will be established and inoculated in nude mice to investigate RAC1/NADPH pathway in the regulation of nasopharyngeal carcinoma cellular radiosensitivity in vivo. The research results will provide a good platform for dynamic monitoring signal transduction pathway of small molecular medicine, screening the lead compound of radiosensitizer and a novel molecular therapeutic target in nasopharyngeal carcinoma.

英文关键词： RAC1/NADPH pathway;anthraquinone compound;radiosensitive;nasopharyngeal carcinoma;Spatial and temporal regulatory