Objective To investigate the effect of andrographolide on head and neck squamous cell carcinoma, and reveal its potential anti-tumor mechanism from the perspective of ferroptophagy through lipidomics. Methods CAL27 cells were cultured in vitro. CCK-8 assay and colony formation assay were used to detect the inhibitory effect of andrographolide on cell proliferation, and flow cytometry was used to detect the cell apoptosis. A patient-derived xenograft (PDX) mouse model of human head and neck squamous cell carcinoma was established. Control group and andrographolide (10 mg/kg) group were set up, and after intervention with administration, the tumor growth was observed dynamically, and the expression of proliferation-related index Ki67 in tumor tissues was detected. Lipidomics technology was used to analyze the metabolic profile changes of CAL27 cells after andrographolide treatment, and Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment analysis was performed. Western blotting was used to detect the expressions of ferroptophagy-related proteins in cells. Results The in vitro experimental results showed that andrographolide could significantly inhibit the proliferation of CAL27 cells and induce cell apoptosis (P < 0.01, 0.001), and showed a dose-dependent effect. The results of in vivo PDX model experiment showed that andrographolide could significantly inhibit the growth of tumors in vivo (P < 0.001), and the positive expression of Ki67 in tumor tissues was significantly reduced compared to the control group (P < 0.001). The lipidomics analysis and KEGG pathway enrichment results showed that the differential metabolic pathways of cells treated with andrographolide were enriched in ferroptosis and autophagy-related pathways. Western blotting results confirmed that andrographolide could significantly regulate the expressions of ferroptophagy-related proteins (P < 0.05, 0.01, 0.001), indicating that andrographolide could effectively induce ferroptophagy in CAL27 cells. Further validation with the nuclear receptor coactivator 4 (NCOA4) targeted degrader and ferroptosis inhibitor demonstrated that both agents markedly reversed the effects of andrographolide (P < 0.05, 0.01, 0.001), verifying that its antitumor effect depends on the ferroptophagy signaling pathway. Conclusion Andrographolide could significantly inhibit the proliferation and induce apoptosis of head and neck squamous cell carcinoma in vitro and in vivo. Its anti-tumor mechanism is related to inducing ferroptophagy in tumor cells.

andrographolide  /  head and neck squamous cell carcinoma  /  ferroptophagy  /  lipomics  /  NCOA4