Background: Liver cancer remains a frequent cause of cancer-related death, and thus targeted drugs urgently need to be developed. Artesunate (ART) inhibits the progression of liver cancer; however, its mechanism of action remains unclear. The primary aim of this study is to clarify whether ART inhibits the progression of hepatocellular carcinoma (HCC) cells by suppressing the Toll-like receptor 4 (TLR4)/ Methods: In vitro studies demonstrated the effects on cell proliferation, invasion, and migration through a series of phenotypic experiments. Specifically, the CCK8 was used to assess the impact on cell proliferation, while the Transwell assay was employed to evaluate the effect on cell invasion. A xeno-inhibitory tumor model was established in vivo to verify the therapeutic effects of ART. Western blotting was used to detect changes in the TLR4/MyD88/NF-xB pathway. Results: The study showed that ART inhibits HCC cell proliferation, invasion, and migration and induces apoptosis in a dose-dependent manner. In vivo studies indicated shown that ART treatment in xenograft tumor models could consistently reduce tumor growth. Moreover, ART inhibited the viability, colony formation, migration, and invasion ability of HCC cells while promoting their apoptosis in a dose-dependent manner. The treatment of xenograft models with ART consistently reduced tumor growth. Furthermore, Western blot analysis demonstrated that the levels of TLR4 and its known downstream effectors (TRAF6, MyD88, and NF-xB) were markedly downregulated after ART treatment in Huh-7 and liposaccharidestimulated Huh-7 cells. Conclusions: These results indicate that ART has a potent effect on the development of HCC cells, the underlying mechanisms of which may be associated with alterations in the TLR4/MyD88/NF-xB signaling pathway in HCC. Therefore, further development of ART as a therapeutic agent is warranted.