<p>Hepatocellular carcinoma (HCC) is a deadly malignancy with poor outcomes. Although there are various treatments for HCC, including surgery, transplantation, and systemic therapy, the overall prognosis remains poor and recurrence after surgical resection is high. Thus, the development of new techniques and new drugs is necessary. In this study, we evaluated the antitumor activity of Atractylon, a bioactive molecule extracted from a Chinese herb of the genus <i>Atractylodes</i>. The results showed Atractylon inhibited proliferation of HCC cells by arresting them at G2/M phase of cell cycle and suppressed their migration and invasion. Using TMT-labeled quantitative proteomics and ferroptosis-related functional assays, we revealed that Atractylon induced HCC ferroptosis by facilitating the accumulation of intracellular Fe²⁺ and reactive oxygen species (ROS), as well as lipid peroxidation. Further study revealed that Atractylon upregulated expression of SLC39A14 (ZIP14), a metal transporter of the hepatocyte membrane. Atractylon-induced ferroptosis and subsequent inhibition of cell proliferation were significantly alleviated following the knockdown of SLC39A14 expression. In summary, this study revealed that Atractylon effectively induced ferroptosis in HCC through the upregulation of SLC39A14 expression and may be a promising candidate for the development of anti-HCC drugs.</p>