Abstract: Objective The aim of this experiment was to investigate effects of schizocapsa plantaginea hance (SSPH), a new anti-cancer natural product, on expressions of ferroptosis-related genes, e.g., glutathione peroxidase 4 (GPX4), solute carrier family 7 member 11 (SLC7A11) and acyl-CoA synthase long-chain family member 4 (ACSL4) in HepG2 and Huh7 cells in vitro. Methods HepG2 and Huh7 cells were respectively divided into control (DMEM), low-dose (0.73 μM), medium-dose (1.46 μM) and high-dose (2.91 μM) SSPH-intervened groups. Cell proliferations were detected by MTT, supernatant Fe²⁺ concentration was determined by o-phenanthroline colorimetry, and reactive oxygen species (ROS) was assayed by fluorescence probe. GPX4, SLC7A11 and ACSL4 mRNA were detected by qRT-PCR and the corresponding protein expressions were detected by Western blot. Results HepG2 and Huh7 cell proliferation greatly in a dose-dependent manner inhibited in low-, medium- and high-dose SSPH-intervened groups as compared to in the control(P<0.05); supernatant ROS and Fe²⁺contents in SSPH-intervened HepG2 and Huh7 cells were significantly increased compared to in the control group (P<0.05);cell GPX4 and SLC7A11 mRNA relative levels in SSPH-intervened HepG2 and Huh7 cells decreased (P<0.05), while ACSL4 mRNA relative level increased greatly compared to in the control (P<0.05); cell GPX4 and SLC7A11 protein expression SSPH-intervened HepG2 and Huh7 cells weaken significantly(P<0.05), while ACSL4 protein expression intensified significantly compared to in the control group (P<0.05). Conclusion SSPH could exert anti-liver cancer effect by inducing ferroptosis in hepatoma cells, and the mechanism by which the intracellular ROS and Fe²⁺ were overloaded, might be related to regulation of ferroptosis-related genes, e.g., ACSL4, GPX4 and SLC7A11 expressions.

Key words: HepG2, Huh7 cells, Schizocapsa plantaginea hance, Glutathione peroxidase 4, Solute carrier family 7 member 11, Acyl-CoA synthase long-chain family member 4, Ferroptosis, In vitro