我们前期研究表明，蟾毒灵能调控肿瘤微环境抑制肝癌侵袭转移，但作用机制有待进一步研究。近年来NAE1蛋白在肿瘤侵袭转移中的作用日益被重视，可介导一系列信号通路调控M1、M2型肿瘤相关巨噬细胞（TAM）浸润。我们前期研究发现，蟾毒灵能下调NAE1蛋白表达。进一步研究发现，肝癌组织中NAE1表达与M2型巨噬细胞浸润正相关。因此，我们提出假说：蟾毒灵通过下调NAE1表达重塑肿瘤微环境中M1、M2型巨噬细胞浸润，发挥抗肝癌侵袭转移作用。本课题以前期研究为基础，通过构建肝癌细胞与巨噬细胞共培养模型、裸鼠尾静脉肝肺转移模型模拟肿瘤微环境，以NAE1及肿瘤微环境中TAM为切入点，并通过TAM芯片筛选并验证表达差异的信号通路，深入解析蟾毒灵抗肝癌侵袭转移的作用环节和调节机制，为以蟾毒灵为组分的中药临床治疗肝癌提供理论依据。

In our previous study, we have demonstrated that bufalin could regulate invasion and metastasis in hepatocellular carcinoma (HCC). However, the concrete mechanisms remain to be explored. The role of NAE1 in tumor invasion and metastasis has been paid much attention to. It has to be reported to modulate M1 and M2 tumor associated macrophage (TAM) via a series of signaling pathways. In our previous study, we have found that bufalin could downregulate NAE1 expression. Further investigation has demonstrated the positive correlation between NAE1 and M2-TAM in HCC. Therefore, we hypothesized that the anti-metastasis effect of bufalin against HCC is regulated by reconstruction of macrophage infiltration mediated by NAE1. Based on previous studies, this subject would establish the models: co-culture of HCC cells with macrophage, liver and lung metastasis mice model via tail vein injection. Besides, TAM chip was employed in the exploration of signaling pathways involved in anti-metastasis effect of bufalin in HCC. This study aims to investigate the mechanisms of anti-metastasis effect of bufalin through exploration of NAE1 and TAM, thus providing theoretical foundation for Chinese medicine’s application in the treatment of HCC.