长非编码RNA LncRNA31两个核质定位不同转录本在肺癌转移中的功能和机制研究

Lung cancer is the leading cause of cancer-related deaths throughout the world in recent years. Once lung cancer has metastasized, that is, spread to other distant organs, the five-year overall survival is less than 5%. Recent advances in whole transcriptome analysis have identified numerous long non-coding RNAs (lncRNAs), which play important roles in a wide range of biological processes and whose deregulation causes human disease, especially in cancer. LncRNAs have been reported to relate to invasion and migration of lung cancer cells, however few reports have been made for the underlying molecular mechanisms. Through both bioinformatics analysis and experimental study, we identified one lncRNA-LncRNA31, decreased in lung cancer, has two different-localization isoforms. Both of the two isoforms inhibited lung cancer cells migration and invasion. Combined RNA pull-down and MS, we found that LncRNA31 isoform1 interacted with VIM and E3 ubiquitin-protein ligase RNF20 and reinforce the interaction between VIM and RNF20. Transport receptor CRM1 mediated LncRNA31 isoform1 export to the cytoplasm. In this study, we want to further elucidate the functions and specific molecular mechanism of LncRNA31 on the basis of the previous research. These results will contribute to the establishment of lncRNA and mRNA regulatory network in lung cancer, and thus the identification of key noncoding sequences in the metastasis of lung cancer, paving the way for finding new targets for tumour diagnosis andtherapy.

近年来，肺癌已成为死亡率最高的恶性肿瘤，肺癌转移病人五年生存率低于5%。研究发现，lncRNA在一系列生物过程中发挥重要作用，其异常表达往往与疾病密切相关尤其是肿瘤。肺癌转移相关的lncRNA已有报道，但具体的分子机制还不清晰。我们运用生物信息学和实验相结合的方法，鉴定出一条肺癌中低表达的lncRNA-LncRNA31，它具有两个亚细胞定位不同的转录本，并且两个转录本都可以抑制肺癌细胞的侵袭迁移。通过RNA pull-down和质谱相结合的方法，我们发现LncRNA31 isoform1与VIM和E3连接蛋白RNF20相互作用。而转运受体蛋白CRM1介导 LncRNA31 isoform1 出核。本课题将在前期研究基础上，对LncRNA31功能和机制进行深入研究，建立肺癌中lncRNA和mRNA的联合调控网络，为肿瘤的诊断和治疗提供新的靶点。

近年来，高通量测序技术的不断改进和广泛应用，极大地加深了我们对基因组非编码序列的认识，这些非编码序列转录出的RNA能够独立地以RNA的形式发挥功能。长链非编码RNA在生命体的各个阶段中都发挥了重要的调控作用，长链非编码RNA的失调导致多种疾病的发生发展，例如癌症、胎儿发育异常、心血管疾病以及神经退行性疾病等。目前，仍有许多长链非编码RNA的功能机制待挖掘和探索。前期我们运用生物信息学和实验相结合的方法，鉴定出一条肺癌中低表达的lncRNA-LncRNA31，它具有两个亚细胞定位不同的转录本，转运受体蛋白CRM1介导 LncRNA31 isoform1 出核，两个转录本都可以抑制肺癌细胞的侵袭迁移。该研究中，我们对两个转录本的具体作用机制进行了详细研究，lncRNA31 isoform1与能够与SERBP1蛋白结合，影响SERBP1蛋白相变，并影响stress granule （SG）形成；isoform2与能够与RNF20蛋白结合，在表观水平调控基因表达。该研究结果为肺癌的诊断和治疗提供新的靶点。在该项目的支持下我们对系统性红斑狼疮中lncRNA-SLEAR和小鼠记忆相关的lncRNA-Carip的功能和机制进行了深入研究，为疾病的发生发展提供了新的治疗靶点。最后在该基金的支持下，我们进行了新冠mRNA疫苗的尝试，初步实验结果显示我们的mRNA疫苗具有一定的通用型，且与国内外现有上市和临床阶段疫苗结果都可比，并具有一定优势。