软肝利胆汤通过miRNA-9-5p调控能量代谢关键酶PDK4抗肝癌作用的机制研究

原发性肝癌严重威胁人类健康。中医药防治肝癌是近年研究的热点。前期临床研究提示软肝利胆汤具有预防术后复发转移、延长生存期、抑制肝癌的作用，但其作用机制尚不明确。丙酮酸脱氢酶激酶4（PDK4）是细胞线粒体能量代谢关键酶，其表达与肝癌预后正相关。预实验结果提示软肝利胆汤可下调人肝癌HepG2细胞miR-9-5p表达，而miR-9-5p的下游靶点为PDK4。因此研究者提出软肝利胆汤通过miR-9-5p调控能量代谢关键酶PDK4抗肝癌的假说。本项目拟采用质粒转染等技术手段，通过细胞培养体外实验和动物模型体内实验，探索软肝利胆汤对miR-9-5p的调控机制，明确miR-9-5p与PDK4的相互作用关系，阐明miR-9-5p/PDK4轴对肝癌细胞线粒体能量代谢及肿瘤增殖、凋亡、迁移和侵袭的影响。本研究发挥中医药优势，丰富中医抗癌理论，为软肝利胆汤抗肝癌的机制提供实验室依据。

Primary liver cancer is a serious threat to human health. The prevention and treatment of liver cancer by traditional Chinese medicine is a hot topic in recent years. Early clinical studies suggested that ruanganlidan decoction has the effects of preventing recurrence and metastasis, prolonging survival period and inhibiting liver cancer, but its mechanism is not clear. Pyruvate dehydrogenase kinase 4 (PDK4) is a key enzyme in mitochondrial energy metabolism, and its expression is positively related to the prognosis of liver cancer. The results showed that ruanganlidan decoction could down regulate the expression of miR-9-5p in HepG2 cells, and the downstream target of miR-9-5p was PDK4. Therefore, the researchers put forward the hypothesis that ruanganlidan decoction can regulate PDK4, the key enzyme of energy metabolism, through mir-9-5p to fight against liver cancer. In this project, we intend to explore the regulatory mechanism of Ruangan Lidan Decoction on mir-9-5p, clarify the interaction between miR-9-5p and PDK4, and clarify the influence of miR-9-5p / PDK4 axis on the energy metabolism of mitochondria and tumor proliferation, apoptosis, migration and invasion of hepatoma cells by using plasmid transfection and other technical means, cell culture in vitro and animal model in vivo. This study gives full play to the advantages of traditional Chinese medicine, enriches the anti-cancer theory of traditional Chinese medicine, and provides the laboratory basis for the anti hepatoma mechanism of ruanganlidan decoction.