MIR34A前体宿主非编码RNA翻译功能性蛋白的发现及在鼻咽癌细胞中作用机理研究

传统认为pri-miRNAs不编码蛋白质，不参与生命进程。前期研究发现，miR-34a前体宿主RNA分子MIR-34AHG可编码多肽，首次命名为MIR34AHG-133aa，通过抗体实验和蛋白质谱数据分析均支持我们的重大发现。细胞功能学研究发现此蛋白能使鼻咽癌细胞周期G1期阻滞，明显抑制癌细胞的增殖。据此提出假设：MIR34AHG可翻译蛋白质参与鼻咽癌细胞的功能调控。本研究拟通过体内外实验，运用质谱技术、Western blot、生物信息学、质粒转染、RNA干扰、CRISPR/cas9基因敲出等技术，阐明miR-34a基因所对应的三种表达产物（miR-34a，MIR-34AHG-133aa及MIR-34AHG）的相互关系以及在鼻咽癌中发挥的作用及分子机制。本项目立意新颖，原创性极强。项目的完成将为基因调控研究开辟新的视野，为鼻咽癌的临床诊治及预后判断提供新靶点和新策略，具有重大意义。

It has been traditionally believed that pri-miRNAs are not involved in protein encoding or other life processes. Our previous research has found that MIR-34AHG, the pri-miRNA of miR-34a, encodes a 133-aa polypeptide (named as MIR34AHG-133aa). These important findings have been confirmed by the antibody detection and protein mass spectrometry analysis. Cell function assessment has shown that MIR34AHG-133aa could induce cell cycle arrest of nasopharyngeal carcinoma cells at the G1 phase, and significantly inhibit the tumor cell proliferation. Therefore, we hypothesize that MIR34AHG-encoded protein participates in regulating the function of nasopharyngeal carcinoma cells. In this study, based on the mass spectrometry, Western blot analysis, bioinformatics analysis, plasmid transfection, RNA interference, and CRISPR/cas9 knockout techniques, the relationship between miR-34a, MIR-34AHG-133aa, and MIR-34AHG, as well as their functions and related mechanisms in nasopharyngeal carcinoma, were investigated, both in vitro and in vivo. Our primary and original findings provide new insights into gene regulation research, and suggest novel targets and strategies for the clinical diagnosis and prognosis of nasopharyngeal carcinoma.