LncRNA AFAP1-AS1上调NONO促进胆囊癌转移的机制研究

Gallbladder cancer is easy to recur and metastasize after operation for the extremely high malignant degree. The long non-coding RNA is differentially expressed in various tumor tissues. The previous study found that lncRNA is differentially expressed in gallbladder carcinoma tissues and participates in the development of gallbladder cancer. Based on our previous studies, we established an in vitro metastasis model and detected metastasis associated lncRNA, and screened lncRNA AFAP1-AS1. Previous study confirmed that silencing of lncRNA AFAP1-AS1 inhibits the metastasis of gallbladder cancer cells, and RNA-pulldown assay showed that lncRNA AFAP1-AS1 binds to NONO. Based on the results, we speculate that lncRNA AFAP1-AS1 binds to NONO, therefore promoting gallbladder cancer metastasis. This project intends to clarify the regulation of AFAP1-AS1 on the metastasis of gallbladder cancer cells; to study the interaction between lncRNA AFAP1-AS1 and NONO, and explore the downstream pathway; to investigate the relationship between lncRNA AFAP1-AS1 and the pathogenesis and prognosis of gallbladder carcinoma. This project is expected to clarify the mechanism of AFAP1-AS1 combined with NONO to promote gallbladder cancer metastasis, and provides a new idea for the diagnosis and treatment of gallbladder cancer.

胆囊癌恶性程度高，易复发转移；长链非编码RNA在各种肿瘤组织中存在差异表达。课题组前期研究发现lncRNA在胆囊癌组织内存在差异表达，参与胆囊癌的发生发展。申请人在前期研究基础上建立了体外转移模型并检测了与转移相关的lncRNA，筛选出一条与转移相关的lncRNA AFAP1-AS1。进一步工作证实敲减AFAP1-AS1可以抑制胆囊癌细胞转移能力，RNA-pulldown发现AFAP1-AS1与NONO结合。据此我们推测AFAP1-AS1与NONO结合后促进胆囊癌转移。本项目拟深入研究:1.明确AFAP1-AS1对胆囊癌细胞转移的调控作用;2.AFAP1-AS1与NONO之间的作用机制，并且探究下游通路;3.探明lncRNA AFAP1-AS1与胆囊癌发病及预后的关系。以期阐明AFAP1-AS1与NONO结合促进胆囊癌转移的机制，为胆囊癌的诊断治疗提供新思路。

胆囊癌是胆道系统中最常见的恶性肿瘤，其恶性程度极高，对放化疗不敏感，术后易于复发转移。长链非编码RNA(LncRNA)是一类长度大于200nt，不具有编码蛋白能力的RNA。其在各种肿瘤组织中存在差异表达，并且在肿瘤发生发展中起到重要作用，因此LncRNA可成为潜在的诊断分子标志物和药物设计的靶点。LncRNA AFAP1-AS1已被证实在肠癌、肺癌等肿瘤的复发转移中中发挥作用。我们通过体内、体外的细胞功能实验证实LncRNA AFAP1-AS1促进胆囊癌细胞的增殖、迁移和侵袭。同时通过RNA-pulldown实验和RIP实验发现LncRNA AFAP1-AS1与NONO直接结合，敲减LncRNA后可以明显下调NONO蛋白和RNA表达水平。因此，我们得出结论，lncRNA AFAP1-AS1在胆囊癌中可以通过结合NONO，达到促进胆囊癌细胞增殖、迁移和侵袭的作用。本项目研究结果将为胆囊癌的诊断和进一步治疗提供新思路和靶标。