FOXD1介导的p53乙酰化在温阳解毒方成分松果菊苷抑制肝癌细胞恶性表型中的机理研究

原发性肝细胞癌（简称肝癌）是最常见的恶性肿瘤之一。许多中药方剂在临床肝癌治疗中疗效显著，但由于有效物质基础和作用机理不明确，推广应用受限。前期研究发现，名中医叶景华经验方温阳解毒方成分松果菊苷显著抑制肝癌细胞的增殖以及FOXD1和TRIB1的表达，且小样本量的肝癌患者中发现FOXD1在癌组织中高表达；申请人已发表论文证实TRIB1与肝癌的恶性表型、p53表达存在相关性。由此我们提出假说，松果菊苷可能通过抑制FOXD1，下调TRIB1表达，从而介导p53乙酰化来调控肝癌细胞恶性表型。本项目拟通过细胞生物学、分子生物学、动物实验及临床样本分析等系统方法，运用基因干扰和过表达、荧光素酶报告基因、染色质免疫沉淀等技术，探讨松果菊苷在肝癌治疗中，通过FOXD1及其下游信号通路对肝癌恶性表型的调控机制。本项目将为验方温阳解毒方治疗肝癌的临床使用提供科学依据，也为肝癌治疗提供新的候选靶点。

Primary hepatocellular carcinoma (HCC) is a kind of common malignant tumor. Many Chinese formulas have been found effective in clinical treatment, but it’s hard for them to be widely acknowledged because of the uncertain material basis and action mechanism. In our previous study, we found that the component echinacoside in Wenyang Jiedu (WYJD) formula from Jinghua Ye could significantly inhibit proliferation of HCC cells and the expressions of FOXD1 and TRIB1, and the expression of FOXD1 was significantly higher in tumor tissues than in non-tumor tissues using a few samples. Furthermore, we have proved that TRIB1 could influence malignant phenotypes of HCC cells and there were some relationships between TRIB1 and p53 in our published paper. Therefore, we propose the hypothesis that echinacoside may influence malignant phenotypes of HCC cells by regulating the expressions of FOXD1 and TRIB1 and the acetylation of p53. This research aims to explore the effect of echinacoside in HCC treatment and clarify the mechanisms of FOXD1 and its downstream signaling pathways regulating malignant phenotypes of HCC cells by gene interference, gene over-expression, luciferase reporter gene assay and chromatin immunoprecipitation assay in levels of cells, molecules, animals and clinic. This research will provide the experimental basis for WYJD formula in HCC treatment, and also contribute new targets for drug development.