华蟾毒配基（cinobufagin）是治肝癌中药蟾酥和华蟾素等的主要活性成分，属于强心苷类固醇物质（CSs)，我们前期首次通过SILAC-LC-MS/MS技术发现，CS可有效抑制肝癌中与细胞纺锤体形成和染色体分离有关的极光激酶A(AURKA)，阻断细胞分裂。而AURKA和ERK信号途径活化可导致肝癌一线药索拉菲尼耐药。我们提出“华蟾毒配基配伍索拉菲尼以AURKA为关键靶点的抑制肝癌细胞增殖并克服耐药“的假说。我们拟敲除或过表达AURKA，应用SILAC-LC-MS/MS筛查AURKA上下游的调控分子，应用生物信息学和分子生物学手段梳理验证以AURKA为节点的信号途径；应用免疫共沉淀和荧光共定位分析通过AURKA介导染色体分离障碍的互作蛋白，阐释染色体分离障碍的机制；应用体外和荷瘤动物实验观察华蟾毒配基配伍索拉菲尼的抗癌作用。研究结果将为明确蟾酥和华蟾素抗癌机理和改善用药提供科学依据。

Cinobufagin is a main active ingredient of traditional Chinese medicine Chansu and cinobufocini and is applied for the treatment of liver cancer. Cinobufagin and plant source of digitalis and ouabain belong to cardiac glycoside steroids (CSs). Our previous studies firstly found that CSs could effectively inhibit the aurora kinase A (AURKA)-mediated spindle formation and chromosome separation signaling pathway, and then block cell division and induce apoptosis in liver cancer HepG2 cells using SILAC and quantitative proteomics strategy. Meanwhile, activation of AURKA signaling is one of the important factors for sorafenib resistance in the process of clinical liver cancer chemotherapy. Hence, we put forward the scientific hypothesis that AURKA as key target for a combination of cinobufagin and sorafenib could inhibit the proliferation of liver cancer cells and overcome the chemotherapy drug resistance of liver cancer cells.In present study, we will screen the up- and downstream molecules of AURKA pathway using the regulation of AURKA expression (knock down or overexpression) and combination of SILAC and proteomics, and validate the critical molecules expression levels associated with AURKA and signaling pathways by bioinformatics and molecular biology strategy. Meanwhile, we will also analyze the anti-liver cancer activity for a combination of cinobufagin and sorafenib in vitro and in vivo experiments. The research results will provide the clear scientific evidences to definite the anti-liver cancer mechanism of Chansu and relevant preparations as well as improving the clinical medications.