长链非编码RNA AK057037在结直肠癌转移中的功能及分子机制研究

Long non-coding RNA (lncRNA) have been confirmed to play critical regulatory roles in cancer biology, but the underlying mechanism remains unknown. AK057037 is a novel lncRNA associated with metastasis of colorectal carcinoma (CRC) by comparing different expression profiles between two CRC cell lines with different metastatic potential using LncRNA microarray in our previous studies. We have identified that AK057037 is associated with metastasis and poor prognosis in CRC by expression analysis and the preliminary function research. In this study, we will evaluate the functions of AK057037 in the metastasis of CRC in vivo and in vitro through RNAi or gene over-expression cell model, and establishment of the whole-body visualization animal models. Furthermore, specific proteins binding with AK057037 will be identified by RNA pull-down, Mass spectrometry and RNA immunoprecipitaiton. Meanwhile, we should screen and identify the key signaling molecules regulated by AK057037 using the gene expression profiling combined with bioinformatics methods. Integrating the function relationships among AK057037, specific binding proteins and the key signaling molecules, we should elucidate the role and molecular mechanism of AK057037 in the metastasis of CRC. Our study findings should Provide new insight into the mechanism of metastasis of CRC and provide new targets for prognosis and preventing metastasis of CRC.

长链非编码RNA(LncRNA) 是一类重要的细胞功能调控分子，研究显示LncRNA参与多种肿瘤的发生、发展进程。AK057037是我们前期应用lncRNA 芯片从不同转移潜能结直肠癌细胞系中筛选获得的与结直肠癌转移相关的一种lncRNA；初步实验证实其与结直肠癌转移及患者预后不良密切相关。本课题拟在前期工作基础上，利用转染、干扰及构建整体可视化动物模型等方法从体内体外两个方面明确AK057037在结直肠癌转移中的作用；进一步应用RNA pull-down 、质谱及RIP筛选鉴定AK057037 特异性结合蛋白；应用基因芯片结合生物信息学方法筛选AK057037 所调控关键信号分子；系统分析结合蛋白和关键信号分子的功能以及与AK057037相互间的功能联系，阐明AK057037 在结直肠癌转移中的功能及作用机制，从lncRNA 这个新视野为阐明结直肠癌侵袭转移的分子机制提供新依据与新思路。

长链非编码RNA(LncRNA) 是一类重要的细胞功能调控分子，在多种肿瘤的发生、发展过程中承担重要角色。课题组应用lncRNA 芯片从不同转移潜能结直肠癌细胞系中筛选获得与结直肠癌转移相关的lncRNA FEZF1-AS1,实验表明其与结直肠癌转移及患者预后不良密切相关,我们应用过表达及干扰技术从体内、体外两方面功能实验证实FEZF1-AS1 的上调可显著增强结直肠癌细胞的增殖、侵袭及转移能力，其下调则明显减低结直肠癌细胞的增殖、侵袭及转移能力；进一步应用RNA pull-down 、质谱分析及RIP 技术，我们筛选获得FEZF1-AS1 特异性结合蛋白FEZF1及POTEE两个重要的功能相关信号分子，并进一步实验证实其二者与FEZF1-AS1的功能联系。实验表明靶基因分子FEZF1 与POTEE 在结直肠癌中高表达，与淋巴结转移、远处转移、Dukes 分期、分化程度正相关，是影响结直肠癌预后的危险因素；功能实验证实FEZF1-AS1可通过靶基因分子FEZF1、POTEE 的介导，参与调控结直肠癌细胞的增殖、侵袭、迁移；而相关机制研究表明POTEE 可通过Rho 信号通路的介导，调控结直肠癌细胞EMT，从而在结直肠癌的增殖、侵袭、转移过程中承担重要作用，本课题的实验结果证实FEZF1-AS1可能为结直肠癌的治疗及预后判断潜在的新分子靶点，从lncRNA 这个新的视野为阐明结直肠癌侵袭转移的相关机制提供了新依据和新思路。