Aurora-A/NF-kappaB/microRNA-17-92基因簇在肝细胞癌恶性生物学行为中的作用及机制研究

Aurora-A, an important member of newly identified serine/threonine kinase family, is found to overexpress in a variety of human cancers, including hepatocelluar carcinoma (HCC). In our previous study, we have shown that the status of Aurora-A expression was significantly correlated with clinical stage, lymph node metastasis and poor prognosis of HCC patients, and silencing of Aurora-A can reverse malignant phenotypes of HCC cells. With the help of previous funds, we have elucidated the upstream molecular mechanisms involved in Aurora-A overexpression in HCC at transcriptional and post-transcriptional levels, but the downstream molecular mechanisms of Aurora-A in HCC are still unclear. On the basis of microRNA microarry results and bioinformatic analyses, we will investigate the molecular mechanisms involved in miR-17-92 cluster-promoting malignant phenotypes of HCC cells dependent of NF-?B signaling mediated by Aurora-A overexpression using a variety of experimental methods (luciferase reporter, gel shift, chromatin immunoprecipitation and point mutation assays, et al). This will be helpful to provide experimental basis for further clarifying and improving the molecular regulatory networks taking the center of Aurora-A gene and exploit a novel therapeutic target in HCC.

Aurora-A是丝氨酸/苏氨酸激酶新家族一个重要分子，课题组前期已对其在肝细胞癌中的临床病理意义、预后价值及其功能进行了相关研究，并在国家自然科学基金资助下分别从转录和转录后水平分析了HIF-1α和miR-129-3p对于Aurora-A表达的上游分子调控机制，但是Aurora-A下游分子调控网络仍不完全清楚。有鉴于此，本课题拟在前期高通量microRNA芯片筛选及生物信息学分析基础上，利用荧光素酶报告基因、凝胶阻滞、染色质免疫共沉淀、位点突变、RNA干扰等方法从细胞和活体动物水平验证Aurora-A基因通过激活核转录因子?B (NF-?B)诱导microRNA-17-92基因簇表达水平升高并最终促进肝细胞癌恶性生物学表型的分子机制，从而为进一步阐明和完善肝细胞癌中以Aurora-A基因为中心的分子调控网络并为发掘肝细胞癌治疗的新靶标提供理论依据。

Aurora-A是丝氨酸/苏氨酸激酶新家族一个重要分子，课题组前期已对其在肝细胞癌中的临床病理意义、预后价值及其功能进行了相关研究，并在国家自然科学基金资助下分别从转录和转录后水平分析了HIF-1α和miR-129-3p对于Aurora-A表达的上游分子调控机制，但是Aurora-A下游分子调控网络仍不完全清楚。有鉴于此，本课题在前期高通量microRNA芯片筛选及生物信息学分析基础上，运用荧光素酶报告基因、凝胶阻滞、染色质免疫共沉淀、位点突变、RNA干扰以及基因过表达等方法验证了Aurora-A基因通过激活核转录因子ĸB (NF-ĸB)诱导microRNA-17-92基因簇表达水平升高并最终转录后水平调控下游两个关键的血管形成相关的靶基因：结缔组织生长因子(CTGF)和血小板反应蛋白-1(TSP-1)。同时，本课题组还分别从体外细胞学和活体动物水平验证了Aurora-A/NF-ĸB/miR-17-92基因簇信号轴参与调控肝细胞癌血管形成以及体内外侵袭转移能力。本研究的完成为进一步阐明和完善肝细胞癌中以Aurora-A基因为中心的分子调控网络并为发掘肝细胞癌治疗的新靶标提供理论依据。