BLACAT2通过结合SF1调控mRNA可变剪接促进膀胱癌细胞上皮-间质转化和转移

项目名称： BLACAT2通过结合SF1调控mRNA可变剪接促进膀胱癌细胞上皮-间质转化和转移

项目编号： No.81472381

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 医药、卫生

项目作者： 何旺

作者单位： 中山大学

项目金额： 78万元

中文摘要： 上皮-间质转化（EMT）是膀胱癌获得转移能力的重要途径，但膀胱癌EMT的调控机制尚未阐明。我们的前期研究发现：长链基因间非编码RNA BLACAT2过表达促进膀胱癌细胞EMT，机制可能是通过结合剪接因子1（SF1）调控mRNA可变剪接。本课题拟进一步：应用免疫印迹、原位杂交等方法在细胞模型、动物模型和临床组织中证明BLACAT2过表达促进膀胱癌EMT和转移，应用RNA免疫沉淀、蛋白质免疫共沉淀等阐明BLACAT2与SF1的直接结合和对SF1功能的调控机制，并应用功能逆转及挽救实验证明BLACAT2对CD44等mRNA可变剪接的调控作用。本项目紧紧围绕BLACAT2直接结合SF1进而调控mRNA可变剪接促进膀胱癌EMT和转移这一科学假说，从BLACAT2的全新调控机制入手，揭示其促进膀胱癌转移的本质，为确立BLACAT2作为膀胱癌分子标记和治疗新靶点提供原创性的科学依据。

中文关键词： 膀胱癌；上皮-间质转化；转移；可变剪接；剪接因子1

英文摘要： Epithelial-mesenchymal transition (EMT) is important in the process of acquiring metastatic ability of bladder cancer cells. But the molecular mechanisms of epithelial-mesenchymal transition in bladder cancer cells remain largely unexplored. Our pilot study have revealed that, overexpression of a long intergenic noncoding RNA (lincRNA) termed Bladder Cancer Associated Transcript 2 (BLACAT2) promotes EMT process of bladder cancer, and possible mechanism may be through binding splicing factor (SF1) and thus modulating massage RNA alternative splicing. In the current study, we will further verify the role of BLACAT2 overexpression in EMT and metastatic process of bladder cancer in cell line model, animal model and clinical tissues, through immunoblot and in situ hybridization, etc. We will further confirm the direct physical association of BLACAT2 with SF1 and its impact on the function of SF1 through RNA immunoprecipitation assay and protein co-immunoprecipitation assay, etc. We will further verify the role of BLACAT2 in modulating mRNA alternative splicing (eg. CD44) through functional reversal and rescue assay. This study will focus on the hypothesis that BLACAT2 promotes bladder cancer EMT and metastasis directly binds SF1 and modulating mRNA alternative splicing and study the novel mechanism of BLACAT2 and thus reveal the mechanistic nature of it in promoting bladder cancer metastasis. Success of this study will provide original scientific basis for BLACAT2 acting as novel biomarker and candidate therapeutic target for bladder cancer in the future.

英文关键词： Bladder cancer;epithelial-mesenchymal transition;metastasis;alternative splicing;splicing factor 1