ABCG2胞间转移瞬时保护肿瘤细胞逃避化疗杀伤的作用机理研究

项目名称： ABCG2胞间转移瞬时保护肿瘤细胞逃避化疗杀伤的作用机理研究

项目编号： No.81503092

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 医药、卫生

项目作者： 王晓坤

作者单位： 中山大学

项目金额： 17.9万元

中文摘要： ABCG2是潜在的肿瘤干细胞标志物。新近，我们发现传统化疗药物可刺激ABCG2从高表达ABCG2的细胞转移至无ABCG2表达的细胞并介导其“瞬时”多药耐药性，但其转移规律及分子机制等均不清楚。本研究拟利用绿色荧光蛋白标记技术，以共聚焦显微镜、流式细胞术等研究药物刺激ABCG2 的胞间转移作用及影响因素，揭示其转移规律；从细胞exosome分泌调控、细胞内吞等方面研究ABCG2的转移机制；利用SILAC蛋白组学结合基因转染技术鉴定参与药物刺激的ABCG2胞间转移的关键蛋白或伴侣分子并阐明其作用机制，为有效克服肿瘤MDR，开发创新型药物及靶向干细胞治疗提供潜在的治疗靶点；利用流式细胞分选技术分选发生ABCG2 转移的敏感细胞，从MDR 表型、干细胞“干性”及稳态维持等方面研究转移的ABCG2功能，为解释肿瘤细胞“瞬时”获得耐药性逃避化疗杀伤提出一种新机理，为寻求新的治疗方案提供新思路。

中文关键词： 多药耐药性；ABC转移泵；乳腺癌耐药蛋白；蛋白质转运；膜蛋白

英文摘要： ABCG2 is a potential biomarker of cancer stem cells. Recently, we found that ABCG2 can be transferred from ABCG2-overexpressing cancer cells to ABCG2-negative recipient cells at the presence of conventional chemotherapeutic agents, resulting in a rapid resistance to chemotherapeutical agents in recipient cells. But the mechanism of intercellular transfer of ABCG2 are poorly understood. To investigate the factors and regularity of the intercellular transfer of ABCG2, the ABCG2 transfer will be imaged by confocal microscopy and determined by flow cytometry at the presence or absence of various factors. The mechanism by which ABCG2 transfers to recipient cells will be investigated, including exosome or endocytosis. Mass spectrometry (MS)-based quantitative proteomics called stable isotope labeling with amino acids in cell culture (SILAC) will be used to indentify and decipher the molecular mechanism or possible pathways involved in the intercellular transfer of ABCG2. The function of transferred ABCG2 will be studied after sorting out the recipient cells by MoFlo XDP Cell Sorter. This study will propose a novel theory to explain why cancer cells can promptly evade chemotherapy. Also, it will provide new potential target or strategy for overcoming MDR mediated by ABCG2 to achieve a better therapeutic effect.

英文关键词： Multidrug resistance;ABC transporter;ABCG2;protein trafficking;membrane protein