基于RNA-Seq筛选的功能性LincRNA-NR_034037与乳腺癌发生发展的关联研究

It had been determined that lincRNAs were associated with a variety of malignancies. In preliminary experiments, we found a significant difference in the expression levels of lincRNA-NR_034037, which is located between TSNAX and DISC1 genes on chromosome 1q42, between breast cancer and surrounding tissues by analyzing our RNA-Seq data from paired breast samples. The fusion mechanism of chimeric transcript TSNAX-DISC1 generated by TSNAX and DISC1 genes was still uncertain. Biochemical analysis further demonstrated thatlincRNA-NR_034037 was associated with the expression of TSNAX-DISC1, and might affect the affinity between insulators and CTCF. Therefore, we hypothesized that lincRNA-NR_034037 might regulate the generation of chimeric transcript TSNAX-DISC1 by cis-splicing of adjacent genes, thereby involving in the development of breast cancer. On the basis of above results, we are to clarify the role of lincRNA-NR_034037 in breast cancer progression with multiple methods, such as two-center large sample analysis, mechanisms exploring, and phenotype-function test, to provide scientific basis and means for screening, prevention, treatment, and prognosis of high-risk breast cancer populations.

研究表明LincRNA与多种恶性肿瘤存在着密切的关系。前期预实验中我们通过RNA-Seq的方法发现1q42染色体上TSNAX和DISC1基因间LincRNA-NR_034037在乳腺癌-癌旁组织中存在差异表达；进一步研究显示该LincRNA与TSNAX-DISC1融合转录本的表达存在关联；目前，TSNAX-DISC1的融合机制尚不明确，而预实验的结果亦提示LincRNA-NR_034037可能参与了调控两基因间绝缘子与CTCF的结合。据此，我们假设该LincRNA可以通过调控顺式剪接（cis-splicing）的方式影响TSNAX-DISC1融合转录本的形成，进而影响乳腺癌的发生发展。本项目拟在此基础上，联合使用双中心大样本分析、机制研究和功能表型实验等方法系统的阐明该LincRNA的异常表达在乳腺癌中的作用。以期为乳腺癌高危人群的筛查、预防、治疗及预后评估提供科学依据和手段。

乳腺癌作为危害我国女性生命健康的主要杀手，其发病率逐年增加。本项目以乳腺癌为对象， 研究RNA-Seq 筛选确定的功能性LincRNA-NR_034037 对该疾病发生及发展的影响。在前期研究的基础上，开展大样本量、双中心的临床样本分析，探讨LincRNA-NR_034037 异常表达与乳腺癌的关系；通过表型检测和一系列体内、外功能表型实验揭示这些遗传改变的功能机制，将宏观关联发现与微观生物学功能统一起来。本项目中我们研究发现了LincRNA-NR_034037能通过竞争性抑制绝缘子上CTCF 结合位点来影响TSNAX-DISC1 融合转录本的形成，从而促进了肿瘤的发生。我们首次研究了LincRNA在调控融合基因形成中发挥的重要作用，为建立乳腺癌高危人群的筛选和预防提供科学依据。