长链非编码RNA与非小细胞肺癌发病风险的关联及其机制研究

The long non-coding RNAs(lncRNAs) as the majority of non-conding RNA, were found to be involved in many biological activity and played a key role in development of many tumors including non-small cell lung cancer (NSCLC). The current findings demonstrated several plasma lncRNAs could be used as a early prediction biomarker for NSCLC. But these lncRNAs were mostly found by comparing the expression of lncRNAs between tumor tissue and tumor adjacent tissue. It is still known little about the role of plasma lncRNAs on NSCLC risk in transcriptome level. To systematic evaluate the value of plasma lncRNAs on the prediction of NSCLC risk, we plan to compare the difference of lncRNAs expression in plasma of before and after accident NSCLC using RNA sequencing based on a prospective cohort, Dongfeng-Tongji Cohort. After selected the NSCLC related lncRNAs by RNA sequencing, the associations between their expression levels and the risk of NSCLC will be tested in a 1 to 3 nested case control study. The positive associated lncRNAs will be further function studied The proposed study will suggest the new biomarker for NSCLC detection and the new mechanism of NSCLC development.

长链非编码RNA(lncRNA)作为非编码RNA中数量最多的RNA，在多种肿瘤的发生发展中扮演了重要角色。现有的研究表明血浆中lncRNA的表达水平与非小细胞肺癌发病风险相关联。但目前所研究的血浆lncRNA多从比较癌和癌旁组织lncRNA表达差异的角度来筛选，该研究策略可能会遗漏较多的血浆lncRNA。为了系统研究血浆lncRNA表达水平与非小细胞肺癌发病风险的关联，本研究依托东风-同济队列，运用转录组测序技术直接比较队列人群中新发非小细胞肺癌病人发病前后血浆中lncRNA表达谱的变化，筛选血浆中潜在的非小细胞肺癌风险相关lncRNA，进而采用巢式病例对照研究设计评估血浆中非小细胞肺癌相关lncRNA表达水平与非小细胞肺癌风险的关联性，并对阳性关联的lncRNA进行功能研究。本研究将为非小细胞肺癌发病风险的早期预测提供新的分子标志物，并为阐明非小细胞肺癌发生发展的机制提供科学依据。

根据本项目的研究目标，我们通过对31对癌和癌旁肺腺癌组织样本进行RNA测序，通过表达差异分析，发现20个lncRNA是肺腺癌相关的lncRNA。利用正常对照和肺癌病人的外周血淋巴细胞进行验证发现，在肺腺癌病人中AP001056.1表达量高于健康对照，而在健康对照中RAMP2-AS1表达量高于肺腺癌病人。因此我们推测AP001056.1表达增加是肺腺癌的危险因素，而RAMP2-AS1是保护因素。进一步的功能分析表明，RAMP2-AS1的靶点在淀粉酶受体复合物2富集最显著，进而确定该lncRNA可通过淀粉酶受体复合物2参与肺腺癌的发生发展。同时，我们利用TCGA数据库肺鳞癌的转录组测序数据，确定4个铁死亡相关的lncRNA AC016924.1、AP006545.2、LINC02555和AL122125.1 是肺鳞癌相关lncRNA。进一步的功能分析表明，这些基因在粘附斑通路富集最显著。表明这些铁死亡相关的lncRNA通过影响粘附斑酶活性来参与肺鳞癌的发生发展。本研究所发现的这些lncRNA可以用于肺腺癌和鳞癌的早期诊断和风险评估。