索拉非尼是治疗晚期肝细胞癌（HCC）最广泛使用的一线药物。不幸的是，索拉非尼耐药往往限制了其治疗疗效。为了评估青蒿素对索拉非尼耐药的HCC的疗效并研究其潜在的药理机制，我们构建了一个“索拉非尼耐药相关基因-ART候选靶点”互作网络，并确定了一个由青蒿素候选靶点AFAP1L2和索拉非尼靶点SRC组成的信号轴，以及下游的FUNDC1依赖性线粒体自噬作为索拉非尼耐药和青蒿素减轻耐药的主要贡献因素。值得注意的是，我们的临床数据表明，HCC组织中AFAP1L2的表达显著高于相邻非癌肝组织（P < 0.05），高表达AFAP1L2还与HCC患者不良总体生存显著相关（P < 0.05）。实验结果显示，AFAP1L2在索拉非尼耐药细胞中过表达，导致下游SRC-FUNDC1信号轴的激活，进一步阻止FUNDC1结合LC3B到线粒体并抑制线粒体自噬的激活，以上结果在体外和体内系统均得到验证。此外，青蒿素通过诱导过度线粒体自噬显著增强了对耐药细胞和肿瘤的索拉非尼的抑制作用。从机械上来说，青蒿素减少了AFAP1L2蛋白的表达，抑制了SRC和FUNDC1蛋白的磷酸化水平，促进了FUNDC1结合大量LC3B到线粒体，进一步过激活了索拉非尼耐药细胞的线粒体自噬和细胞凋亡。总之，青蒿素可能是减轻HCC中索拉非尼耐药的一种有前景的策略，通过加重AFAP1L2-SRC-FUNDC1轴依赖的线粒体自噬。

Sorafenib is the most widely used first-line drug for the treatment of the advanced hepatocellular carcinoma (HCC). Unfortunately, sorafenib resistance often limits its therapeutic efficacy. To evaluate the efficacy of artesunate against sorafenib-resistant HCC and to investigate its underlying pharmacological mechanisms, a "sorafenib resistance related gene-ART candidate target" interaction network was constructed, and a signaling axis consisting with artesunate candidate target AFAP1L2 and sorafenib target SRC, and the downstream FUNDC1-dependent mitophagy was identified as a major contributor to the sorafenib resistance and a potential way of artesunate to mitigate resistance. Notably, our clinical data demonstrated that AFAP1L2 expression in HCC tissues was markedly higher than that in adjacent non-cancerous liver tissues (P < 0.05), and high AFAP1L2 expression was also significantly associated with an unfavorable overall survival of HCC patients (P < 0.05). Experimentally, AFAP1L2 was overexpressed in sorafenib resistant cells, leading to the activation of downstream SRC-FUNDC1 signaling axis, further blocking the FUNDC1 recruitment of LC3B to mitochondria and inhibiting the activation of mitophagy, based on both in vitro and in vivo systems. Moreover, artesunate significantly enhanced the inhibitory effects of sorafenib on resistant cells and tumors by inducing excessive mitophagy. Mechanically, artesunate reduced the expression of AFAP1L2 protein, suppressed the phosphorylation levels of SRC and FUNDC1 proteins, promoted the FUNDC1 recruitment of massive LC3B to mitochondria, and further overactivated the mitophagy and subsequent cell apoptosis of sorafenib resistant cells. In conclusion, artesunate may be a promising strategy to mitigate sorafenib resistance in HCC via exacerbating AFAP1L2-SRC-FUNDC1 axis-dependent mitophagy.