circRARS促进非小细胞肺癌恶性进展的功能和机制研究

An increasing attention has been paid on the role of circular RNA in cancer recently. We have identified a high expressed circular RNA with poor prognosis in non-small cell lung cancer patinets by high-throughput sequencing and qRT-PCR, named circRARS. Silence of circRARS could inhibit proliferation and invasion of lung cancer cells. Our previous study showed circRARS is located in nucleus with a miR-634 binding site, and can bind with Ago2 protein. circRARS could co-expression with ZEB2, a potential target gene of miR-634. We hypothesize that circRARS could binding with miR-634, and upregulate ZEB2 expression through ceRNA machanism. In the current proposal,we aim to confirm circRARS could promote NSCLC malignant progression, and verify our hypothesis through luciferase reporter gene and RIP methods, et al. The function and molecular mechanism of circRARS has not been reported so far, therefor, the current study is of potential novelty and might provide new insights for the prevention and treatment of NSCLC.

环状RNA在肿瘤中的作用日渐被研究重视，申请人通过去核糖体测序和qRT-PCR鉴定出一个在NSCLC中高表达且与患者预后不良相关的环状RNA，circRARS，敲减circRARS可抑制细胞增殖与侵袭能力。我们的前期工作表明circRARS分布于细胞核、含有miR-634结合位点、能与Ago2蛋白结合、与miR-634潜在靶基因ZEB2共表达；因此提出“circRARS可吸附miR-634，通过ceRNA机制上调ZEB2表达、促进NSCLC恶性进展”的机制假说。本课题将在前期研究基础上进一步验证circRARS促进NSCLC恶性进展的生物学功能，通过荧光素酶报告基因、RIP等实验阐明circRARS吸附miR-634上调ZEB2的机制假说。目前尚未见有关circRARS功能与机制的研究，本项目将为NSCLC防治提供新的思路。

环状RNA hsa_circ_0001551（circRARS）在非小细胞肺癌（NSCLC）肿瘤组织和细胞中均是过表达的。结合临床与预后信息，circRARS与高TNM分期和淋巴结转移具有显著的相关性，同时具有高表达circRARS的NSCLC病人通常具有较短的生存时间。利用siRNA和过表达质粒检测表型实验，circRARS促进非小细胞肺癌细胞的增殖、侵袭和迁移。机制实验上，利用核质分离实验发现circRARS只要分布于细胞质中，进一步通过RNA pull-down联合质谱分析，circRARS可能结合乳酸脱氢酶（LDHA）。LDHA是有氧糖酵解过程产生乳酸的最后一个关键酶，体外实验证明circRARS可以在转录水平调节LDHA的表达以及调节LDHA的活性。随后，通过检测葡萄糖消耗量和乳酸生成量发现circRARS可以促进有氧糖酵解。综合，circRARS在非小细胞肺癌中作为癌基因可以作为潜在的治疗靶点。本项目资助发表SCI文章2篇，培养博士后1名，博士生4名（其中2人已毕业），硕士生2名。本项目获得资助25.2万元，支出24.911万元，结余0.289万元，基本符合预算。