沉默ARK5基因逆转乏氧诱导胃癌多药耐药的机制研究

项目名称： 沉默ARK5基因逆转乏氧诱导胃癌多药耐药的机制研究

项目编号： No.81460371

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： 易波

作者单位： 江西省肿瘤医院

项目金额： 47万元

中文摘要： 多药耐药(MDR)导致胃癌化疗失败，乏氧是MDR产生的重要原因，发现并调控乏氧诱导胃癌MDR的关键分子可以克服胃癌耐药性。本课题组发现新辅助化疗无效患者ARK5及其磷酸化活化状态（p-ARK5）呈强阳性，最新研究报告ARK5是细胞内 能量监控分子，在乏氧条件下重新调节能量代谢，使细胞幸免。结合前期工作，我们假设：ARK5在乏氧所致的MDR中发挥重要作用，其机制是乏氧通过HIF-1α调控ARK5表达，激活Akt/ARK5信号通路，p-ARK5在14-3-3ζ分子向导作用下由胞浆迁移至线粒体，实现能量调控作用。本课题拟构建低分化人胃癌耐药细胞株，通过体内及体外实验，分别在常氧及乏氧条件下，采用腺病毒载体构建、质粒转染、RNA干扰、Co-IP、双标记免疫荧光法等技术，系统地阐明ARK5及其信号传导通路在乏氧诱导胃癌细胞MDR中的机制，为开发ARK5分子靶向治疗药物奠定理论基础。

中文关键词： C07_胃肿瘤；多药耐药；AMPK的相关蛋白激酶5；低氧诱导因子-1α；14-3-3ζ

英文摘要： The multidrug resistance（MDR）is the basic cause of failure of chemotherapy in patients with gastric cancer,and hypoxia is one of the important cause of MDR.The resistance could be overcome by regulating the key molecules in multidrug resistance induced by hypoxia.Our research group has found in patients with invalid neoadjuvant chemotherapy the ARK5 and it's phosphorylated activation state ( p-ARK5 ) was strongly positive, and it was reported recently ARK5 was an important intracellular energy monitor , which regulated energy metabolism in hypoxic conditions and made the cell survive. Combined with the previous work, we put forward the following hypothesis: ARK5 play an important role in MDR induced by hypoxia through the mechanism that under the hypoxic condition HIF-1α regulated the ARK5 transcription, and activated Akt/ARK5 signaling pathway, p-ARK5 translocated from endochylema to chondrosome by the conduction fo 14-3-3 zeta, finally achieved energy regulation.This project aims to systematicly elucidate the mechanism of ARK5 and its signal transduction pathway in hypoxia induced gastric carcinoma MDR by building low differentiation human gastric cancer drug-resistant cell line,using the adenovirus vector, plasmid transfection, RNA interference, Co-IP, double labeling immunofluorescence technique, in vivo and in vitro, respectively under normoxic and hypoxic conditions. And further to provide the theoretical foundation for the molecular targeted therapy aimed at ARK5.

英文关键词： stomach neoplasm;multidrug resistance;AMPK-related kinase-5;Hypoxia induced factor-1α;14-3-3ζ