miRNA-885-5p抑制原发性肝细胞癌生长与转移机制研究

Our previous study found that there is a close correlation between the level of circulating miR-885-5p and the progress of liver-related pathologies including CHB (chronic hepatitis B), LC (liver cirrhosis) and HCC (hepacellular carcinoma).Our previous study found for the first time that miR-885-5p could serve as novel complementary biomarkers for the detection and assessment of liver pathologies.In addition,bioinformatics and our primary experiments show that miR-885-5p can directly target b-catenin and TCF4, which suggests miR-885-5p is a negative regulator of HCC. In this project, we plan to investigate the connection between mi RNA-885-5p expression and degrees of malignancy in hepatoma as well as the role of mi RNA-885-5p in regulating wnt signal pathway. The project intends to test and verify the hypothesis that miRNA-885-5p could suppress hepatoma growth and metastasis and explore the regulating mechanism of miRNA-885-5p in wnt pathway. And then we would examine the anti-tumor effects of targeted nanoparticles loaded with miRNA-885-5p in NOD/SCID xenograft models by in vivo imaging technique. Together, our study will suggest that miR-885-5p is a negative regulator of HCC metastasis and indicate a potent rationale for developing miR-885-5p as a potential therapeutic agent against HCC metastasis.

本课题组前期工作发现miRNA表达谱异常与肝炎、肝硬化向原发性肝细胞癌（HCC）的发展密切相关，首次发现miRNA-885-5p可作为新的潜在辅助诊断肝脏疾病标志物。生物信息学预测及初步实验显示，它可调控Wnt通路中重要辅转录因子β-catenin和TCF4，提示miRNA-885-5p是一种新的潜在防治HCC发生的调控因子。本课题拟通过探索miRNA-885-5P与肝细胞癌恶性程度的关系及其在体内外对肿瘤细胞增殖及转移的影响，进而通过建立小鼠肝癌自发性转移模型，利用体内分子成像技术研究miRNA-885-5p纳米靶向颗粒的抗肿瘤与抑制肿瘤转移的功效。我们的研究结果将为miRNA对抑制肝癌生长及转移的作用提供理论依据，为肝癌基因诊断、发生机制及治疗靶点的研究奠定更多研究基础。

本研究探索了miR-885-5p 在肝癌转移中的作用。表达研究发现miR-885-5p在不同临床分期的肝癌组织和不同恶性程度的细胞系中表达具有明显差异。在细胞系中，转移能力较弱miR-885-5p表达水平较高，而在组织标本中，临床分期较早表达水平较高，并且高表达miR-885-5p 的病人预后较好。进一步进行了功能实验，在低表达miR-885-5p 的肝癌细胞系中过表达miR-885-5p，在高表达miR-885-5p 的肝癌细胞系中沉默miR-885-5p，采用Transwell 和划痕实验检测miR-885-5p 对细胞系迁移和浸润的影响，发现过表达miR-885-5p 可以抑制肿瘤的转移，沉默miR-885-5p 可以增强细胞的增殖。肝癌原位成瘤模型实验结果表明，过表达miR-885-5p也可以在体内抑制肝癌的转移，并与小鼠的生存期呈正相关。 . 因此，本研究结果表明miR-885-5p 的表达和HCC 的临床分期呈负相关，并且影响患者预后；miR-885-5p 在体内体外均可以抑制HCC 的生长和转移。本研究可以为未来HCC潜在的治疗方法提供理论依据。..