lncRNA DLEU1/miRNA-124-3p/MOV10调控网络诱导EMT影响胶质瘤替莫唑胺耐药的机制研究

我们前期研究发现lncRNA DLEU1可通过调控上皮间质转化(EMT)中Akt/GSK-3β/β-catenin信号通路及相关蛋白的表达介导胶质瘤替莫唑胺的耐药，但作用机制尚不明确。本项目致力于深入探究lncRNA DLEU1的两种作用模式：RNA结合蛋白与ceRNA作用模式，通过生物信息学预测与体外实验，初步查明lncRNA DLEU1/miRNA-124-3p/MOV10可形成环形调控网络；拟采用FISH、RNA-pulldown、RIP、激光共聚焦、双荧光报告基因等实验，揭示此调控网络的存在，并确定上下游调控关系及具体结合位点。最终，在细胞水平、动物水平、临床水平系统阐明这一调控网络通过诱导EMT相关蛋白的表达，介导胶质瘤替莫唑胺耐药的作用机制，明确lncRNA DLEU1在此调控网络中的重要作用，为胶质瘤替莫唑胺的个体化用药提供潜在的生物标志物。

Our previous work indicated that lncRNA DLEU1 mediates the resistance of temozolomide in glioma by regulating the Akt/GSK-3β/β-catenin signaling pathway and related proteins of epithelial-mesenchymal transition (EMT), while the mechanism is still unclear. This research aims to further explore two modes of lncRNA DLEU1 action: RNA binding protein and ceRNA. It was found that lncRNA DLEU1/miRNA-124-3p/MOV10 can form a circular regulatory network through bioinformatics prediction and cell experiments. Fish, RNA-pull down, RIP, laser confocal and dual-fluorescence reporter gene will be proposed to reveal the existence of this regulatory network, further identify the regulatory relationship of upstream and downstream and the specific binding sites. In a word, the mechanism of this regulatory network of mediating the resistance of temozolomide in glioma via regulating the related proteins of EMT will comprehensively demonstrated in cells, animals and clinical levels. The importance of lncRNA DLEU1 in this regulatory network will be clarified, which provides potential biomarkers for the individualized use of temozolomide for glioma.