截断型APC蛋白结合lncRNA-APC2促进结直肠癌恶性进展的作用及机制

结直肠癌（CRC）是我国最常见的恶性肿瘤之一，而APC基因突变是CRC发生发展的关键引发步骤。多项研究表明：绝大部分APC基因突变会产生截断型APC蛋白，且此类型蛋白对CRC恶性进展发挥重要的促进作用，然而其分子机制不明确。最近，我们通过RIP-seq实验对常见截断型APC蛋白的互作RNA分子进行筛选鉴定，并通过体内外功能实验确定了一个与截断型APC蛋白结合并介导其功能的长链非编码RNA（lncRNA-APC2）。但截断型APC蛋白与lncRNA-APC2互作的具体机制不清楚。本项目将进一步通过RACE、FISH、RIP、RNA pull-down、CLIP-seq、IP-质谱鉴定、表达谱测序等技术及体内外功能实验及临床标本验证明确lncRNA-APC2分子特征、及其与截断型APC蛋白互作的复合物成分、关键结构域、临床意义及潜在转化价值。本研究将为揭示CRC恶性进展机制及治疗提供新线索。

Colorectal carcinoma (CRC) is one of the most common human malignancies in China. Mutations in the gene adenomatous polyposis coli (APC) play a pivotal role in tumourigenesis and progression of CRC. Multiple studies have shown that truncated APC protein play an important role in promoting CRC progression, but the underling mechanism remains unknown. Recently, we performed RIP-seq assay to screen the binding RNAs with classic truncated APC proteins. Furthermore, we identified a long non-coding RNA (lncRNA-APC2) mediating the function of truncated APC through in vitro and in vivo function experiments. However, the specific mechanism is not clear. In this project, we will further investigate and confirm the molecular characterization of lncRNA-APC2, and the members、key domain、clinical significance and potential value of the complex through a series of molecular biological assay such as RACE、FISH、RIP、RNA pull-down、CLIP-seq、IP-Mass spectrometry、expression profile sequencing and in vitro and in vivo function experiments. This study will provide new clues for revealing the mechanism of CRC progression and therapy strategy.