Introduction: Shikonin, a traditional Chinese medicine monomer, exhibits potential anticancer activity. An expanding body of literature indicates that targeting ferroptosis signaling pathways can elicit antitumor effects in HCC. However, it remains unclear whether shikonin inhibits HCC by inducing ferroptosis. This study aimed to verify the anticancer effect of shikonin on HepG2 cells and elucidate its underlying mechanisms and specific therapeutic targets through an integrative approach combining network pharmacology, bioinformatics, and experimental validation. Methods: HCC, shikonin targets, and ferroptosis-related genes were retrieved from public databases and their intersection genes were identified. PPI network and enrichment analysis were used to identify them, and "cellular response to metal ion" was a highly enriched GO biological process. Molecular docking was conducted to evaluate the binding affinity between shikonin and candidate targets, followed by in vitro validation. We also constructed a prognostic nomogram via multivariate Cox regression to estimate 1-, 3-, and 5-year overall survival. Results: We obtained a total of 12 intersecting genes. AURKA, identified as a hub gene, showed direct binding to shikonin via molecular docking, pointing to its therapeutic targeting potential for shikonin in HCC. Validation experiments showed shikonin induced ferroptosis and inhibited proliferation in HepG2 cells, accompanied by AURKA downregulation at both mRNA and protein levels. After prognostic model construction, validation experiments confirmed that the nomogram accurately predicted survival outcomes in HCC patients. Discussion: This study explores the therapeutic potential of shikonin in inducing ferroptosis for treating hepatocellular carcinoma (HCC). Through network pharmacology, molecular docking, and in vitro assays, AURKA was identified as a key target, demonstrating that shikonin inhibits HCC cell proliferation by triggering ferroptosis, which suggests a promising ferroptosis-based treatment strategy for HCC. Conclusion: Together, shikonin exerts significant antitumor effects on HepG2 cells, and the underlying mechanism may involve targeting AURKA to induce ferroptosis.
Zhang et al. (Mon,) studied this question.