Wnt/β-catenin信号通路介导的敷和备化方对肝癌干细胞作用机制的研究

Hepatic cancer stem cells are recognized by most scholars as the origin of the hepatic cancer and recurrence after the treatment. Inducing hepatic cancer stem cells differentiation will give more rise to hepatic cancer targeted therapy. Fuhe Beihua Decoction is the clinical prescription experienced by the national old Chinese medicine professor Rong Yuanming. The research bases on the previous study which confirmed Fuhe Beihua Decoction could significantly improve hepatic fibrosis and lower AFP in hepatic cancer patients. Under the instruction of TCM theory on "liver and spleen", research group thinks Fuhe Beihua Decoction can reverse induce hepatic cancer stem cells involving of Wnt/β-catenin signal pathway in FuHe BeiHua Decoction. The research will explore the relationship between hepatic cancer stem cells differentiation reverse induced by Fuhe Beihua Decoction and significance of Wnt/β-catenin based on the level of cells and animals. Using in vitro cell culture technology, the determination of cell survival rate through CCK 8, ELISA, qRT-PCR and Western Blot to detect Wnt/β-catenin signal pathway related gene and protein expression. To certify the mechanism of hepatic cancer stem cells differentiation induced by Fuhe Beihua Decoction and provide theoretical and experimental basis for hepatic targeted therapy in the future.

肝癌干细胞作为肝癌发生及治疗后复发的起源被多数学者认可。诱导肝癌干细胞逆向分化为肝癌靶向治疗开辟了新思路。敷和备化方是全国名老中医荣远明教授的临床验方。本课题组以前期研究证实敷和备化方可明显改善肝癌患者肝纤维化及降低AFP的事实为基础，在中医“肝脾理论”指导下，提出了敷和备化方可在Wnt/β-catenin信号通路的介导下诱导肝癌干细胞逆向分化的假说。本项目从细胞及动物水平上，探索敷和备化方诱导肝癌干细胞逆向分化情况及分化过程中与Wnt/β-catenin信号通路介导的相关性。运用体外细胞培养技术、通过CCK-8测定细胞存活率，ELISA、qRT-PCR、Western Blot检测Wnt/β-catenin信号相关基因及蛋白的表达，验证敷和备化方诱导肝癌干细胞分化作用和机制，为未来肝癌的临床靶向治疗提供理论和实验依据。

肝癌干细胞作为肝癌发生及治疗后复发的起源被多数学者认可。诱导肝癌干细胞逆向分化为肝癌靶向治疗开辟了新思路。敷和备化方是全国名老中医荣远明教授的临床验方。本课题组以前期研究证实敷和备化方可明显改善肝癌患者肝纤维化及降低AFP的事实为基础，在中医“肝脾理论”指导下，提出了敷和备化方可在Wnt/β-catenin信号通路的介导下诱导肝癌干细胞逆向分化的假说。本项目从细胞及动物水平上，探索敷和备化方诱导肝癌干细胞逆向分化情况及分化过程中与Wnt/β-catenin信号通路介导的相关性。运用体外细胞培养技术、通过CCK-8测定细胞存活率，ELISA、qRT-PCR、Western Blot检测Wnt/β-catenin信号相关基因及蛋白的表达，验证了敷和备化方能抑制肝癌干细胞自噬和增殖，其机制与抑制PI3K/AKT/mTOR信号通路有关；能有效降低肝癌干细胞表面标记物 CD133和干细胞转录因子SOX2、NANOG、OCT4的表达水平，降低肝癌干细胞恶性程度。