Artematrolide F suppresses communication between hepatocellular carcinoma cells and hepatic stellate cells to attenuate liver cancer progression through targeting YKT6.

Hepatocellular carcinoma (HCC) remains a highly malignant cancer with limited treatment options. HCC cells (HCCs) activate hepatic stellate cells (HSCs) and transform them into cancer-associated fibroblasts (CAFs), forming a vicious cycle for hepatocarcinogenesis. Therefore, inhibition of the communication between HCCs and HSCs is expected to emerge as a novel strategy for HCC treatment. Artematrolide F (AF), a sesquiterpenoid dimers (SDs) isolated from Artemisia atrovirens, exhibited significantly inhibitory activity on tumor cells in our previous report, but its mechanism remains unknown. This further study demonstrated that AF significantly inhibited the proliferation, colony formation and metastasis of HCCs. Notably, the inhibitory effect was enhanced when HCCs were cocultured with HSCs, suggesting that AF may suppress tumor progression through regulating the crosstalk of HCCs and HSCs. The vesicular transport protein YKT6 was identified as a target of AF through integrating bioinformatics analysis and machine learning model, which were verified by DARTS, CETSA, molecular docking, molecular dynamics simulation, and knockdown experiments. Further mechanistic research revealed that AF targeting YKT6 disrupted the crosstalk between HSCs and HCCs, suppressed HSC-driven tumor proliferation by inhibiting extracellular vesicles (EVs) secretion from HCCs, and blocked tumor autophagy by disrupting vesicular transport. These findings demonstrate that AF could serve as a novel therapeutic candidate for HCC treatment through regulating YKT6 to suppress HCCs-HSCs communication, block EV-dependent tumor progression, and inhibit tumor autophagy.