The clone formation assay also reflected the attenuation of curcumin's ability to inhibit long-term proliferation of hepatocellular carcinoma cells after knockdown of ACSL4 (Fig. 4H).This suggests that the level of defense against ferroptosis in hepatocellular carcinoma cells was significantly increased after silencing ACSL4 and mainly involved the cystine/glutathione/glutathione peroxidase 4 signaling pathway axis. This evidence concerns the gene GPX4 and hepatocellular carcinoma.