LncRNA RP1-13P20.6调控Vav1基因抑制结直肠癌细胞增殖及侵袭效应 的机制研究

LncRNA is a kind of non-coding RNA with regulatory function. We found that lncRNA RP1-13P20.6 was down-regulated in colorectal cancer tissues and inhibited the proliferation and invasion of colorectal cancer cells. Using transcription-associated trap, we found that oncogene Vav1 was one of the targets of RP1-13P20.6. The transcription and translation level of Vav1 was decreased significantly when RP1-13P20.6 was overexpressed. Bioinformatics analysis showed that transcription factor C/EBP-β had multiple binding sites with promoter region of Vav1 gene and C/EBP-β was one of the target proteins of RP1-13P20.6. Therefore, we hypothesis that RP1-13P20.6 inactivated transcription factor C/EBP-β and down-regulated the expression of Vav1 gene, and then inhibited the proliferation and invasion of colorectal cancer cells. We aim to verify the mechanisms of RP1-13P20.6 in regulating Vav1 and the effects on oncogenesis and the development of colorectal cancer in molecular, cellular, animal and clinical level, and try to explore the molecular mechanisms of how RP1-13P20.6 suppress tumor growth. This will further enrich the theory of the regulatory roles of lncRNAs participating in multiple diseases and may provide a new target for the treatment of colorectal cancer.

LncRNA在多种疾病中发挥重要调节作用，申请者发现并报道lncRNA RP1-13P20.6在结直肠癌组织中异常低表达并发挥抑癌作用。进一步研究结果证实RP1-13P20.6靶向结合Vav1基因启动子区，发挥负向调节作用。生物信息学分析显示转录因子C/EBP-β与Vav1基因启动子区有多个结合位点，且RP1-13P20.6含有C/EBP-β的多个反应元件。因此我们推测RP1-13P20.6介导转录因子C/EBP-β失活、抑制Vav1基因表达，进而抑制结直肠癌细胞的增殖与侵袭。本研究拟利用RIP实验、双荧光素酶实验及裸鼠成瘤等在分子、细胞、动物及临床水平验证RP1-13P20.6对Vav1的调控作用及其对结直肠癌发生发展的影响，探索RP1-13P20.6发挥抑癌作用的分子机制，进一步丰富非编码RNA参与疾病调控的理论研究，并可能为结直肠癌的治疗提供新靶点及理论基础，具有一定科学意义。

LncRNA在多种疾病中发挥重要调节作用，本项目研究发现lncRNA RP1-13P20.6通过负向调控vav1基因进而抑制结直肠癌细胞的增殖和侵袭。进一步的研究证实了lncRNA RP1-13P20.6与转录因子C/EBP-β的直接结合作用。同时，我们通过质谱分析发现有427个蛋白在lncRNA RP1-13P20.6过表达后发生表达差异，显著下调的蛋白包括DNAJB12、ENO1、CDK6、CDK4、HK2、S100A4、IFITM家族中的IFITM3、IFITM2及IFITM1等，通过GO分析及有氧糖酵解相关实验我们发现lncRNA RP1-13P20.6主要通过调控结直肠癌细胞有氧糖酵解而发挥抑癌作用。另外，我们针对IFITM家族的IFITM10进行了进一步研究，结果发现IFITM10在胃肠癌的癌组织中表达上调，并发挥促癌作用。本课题组在前期工作基础上，探索RP1-13P20.6发挥抑癌作用的分子机制，进一步丰富了非编码RNA参与疾病调控的理论研究。