表观遗传学异常是人类恶性肿瘤发生进展的重要原因，tRNA的m6A甲基化修饰在tRNA的剪接中发挥重要作用，tRF-RNA来源于tRNA精确剪切产物，tRF-RNA通过靶向竞争性结合RNA稳定蛋白，发挥“置换机制”直接或间接调控肿瘤细胞的生物学行为。本项目拟在课题组前期细胞及组织预实验和相关生物信息学分析的基础上，运用LC-MS/MS、CLIP、RIP-seq、RNA-EMSA、小动物活体成像等技术，在细胞学、动物模型、临床样本三个层次，逐层深入验证tRNAVal去除m6A修饰后剪切产生tRF-RNA3008A，它替代促癌基因ADAMTS-4 mRNA，竞争性结合RNA稳定蛋白Ago2，导致ADAMTS-4 mRNA发生降解，从而抑制结肠癌细胞增殖和侵袭转移的科学假设及分子机制。为靶向m6A修饰酶、tRF-RNA、RNA结合蛋白，改善结肠癌患者预后，提供新的线索和可靠的科学理论依据。

Epigenetic abnormality is an important root of the invasion and metastasis of colon cancer. The m6A methylation of tRNA plays an important role in the splicing of tRNA. tRF-RNA is generated by transfer RNA via m6A demethylation, which regulate the biological behaviors of malignant tumor cells by targeting RBP. Based on our preliminary experimental discoveries gotten from clinical tissue and human cell lines, as well as bioinformatics analysis for these preliminary results, the current project aims to prove our scientific hypothesis: Following m6A demethylation，tRNAVal forms tRF-RNA3008A through specific cleavage. tRF-RNA3008A competitively combines with a RNA stabilization protein of Ago2, which is originally bound with an oncogenic gene of ADAMTS-4 mRNA. Without a binding with Ago2, ADAMTS-4 mRNA will become unstable and easy to be degraded. As a result, tRF-RNA3008A will inhibit the proliferation and metastasis abilities of colon cancer cells through this replacement mechanism by a competitive binding to Ago2, instead of original RNA binder of ADAMTS-4 mRNA. The present study will prove our scientific hypothesis in independent experiments on cytology, animal models and clinical specimens by the application of multi-techniques including LC-MS / MS, CLIP，RIP-seq, RNA-EMSA, and in vivo imaging in small animal. The findings achieved from the present project are supposed to elucidate a novel formation mechanism of tRF-RNA and its anti-carcinogenic roles in colon cancer. It may also provide us with new clues and experimental evidences to discover new therapeutic vectors to improve the prognosis of colon cancer patients by targeting m6A modification enzymes, tRF-RNAs, RNA binding proteins from the viewpoint of epigenetics.