基于课题1筛选到具有调控肿瘤转移作用的lncRNA，本课题从lncRNA调控肿瘤转移信号传导入手探索其分子机理。通过干预lncRNA表达，高通量筛选和目标鉴定激活的蛋白信号通路，探索lncRNA对肿瘤转移信号传导的调节作用，并观察信号通路激活对lncRNA表达的调控，解析lncRNA与蛋白质形成的信号调控网络。通过RNA-Chip和RNA-EMSA等技术探索lncRNA调控信号通路蛋白转录的机制，通过RNA免疫共沉淀等分析lncRNA与信号通路蛋白结合并调节其功能的作用。通过光增强核苷酸交联和免疫沉淀等技术确定lncRNA与蛋白结合的特定序列，通过质谱辅助的生物素标记等方法明确蛋白质与lncRNA结合的氨基酸序列。在细胞培养和荷瘤鼠中探索lncRNA通过调控肿瘤细胞信号传导对肿瘤转移的影响。本研究将为揭示lncRNA调节肿瘤转移信号传导的新机制，解析其与蛋白信号通路的交联调控网络奠定基础。

Based on screenings for functional lncRNAs regulating tumor metastasis by sub-project 1, we will explore the molecular mechanisms of these lncRNAs by studying how they regulate metastasis-associated signal pathways. By changing the expression of lncRNAs, we will screen for and identify the affected signal pathways, explore the roles of lncRNAs in regulating metastasis-associated signal pathways, and observe the regulation of lncRNAs by these signal pathways backwards, so that we can figure out a lncRNA-protein signal network. We will then explore the mechanisms by which lncRNA regulating transcription of the signaling proteins through RNA-ChIP and RNA-EMSA; analyze the role of lncRNAs to interact with and regulate the functions of the signaling proteins by RNA-IP; identify the sequence of lncRNA responsible for specific recognition of interacting proteins by PAR-CLIP; identify motif of proteins responsible for specific recognition of interacting lncRNAs by MS-aid biotin labeling assays. Finally, we will explore the metastasis-regulating functions of lncRNAs by regulating signal pathways. By carrying out this sub-project, we will be able to discover novel mechanisms by which lncRNA regulates metastasis-associated signal pathways, and to understand the lncRNA-protein signal networks.