Bavachin stimulates ferroptosis and reduces malignant phenotype progression of hepatocellular carcinoma cells by inducing lipid peroxidation by modulation of the Nrf2/HO-1 signaling pathway

Haoyu Li; Jianyong Yuan; Wei Dong; Cheng Yang; Lihua Lu; Yizhou Wang

doi:10.62347/HAEU6139

Bavachin stimulates ferroptosis and reduces malignant phenotype progression of hepatocellular carcinoma cells by inducing lipid peroxidation by modulation of the Nrf2/HO-1 signaling pathway

Am J Transl Res. 2024 Nov 15;16(11):6925-6934. doi: 10.62347/HAEU6139. eCollection 2024.

Authors

Haoyu Li¹, Jianyong Yuan², Wei Dong³, Cheng Yang⁴, Lihua Lu^{5

6}, Yizhou Wang^{5

6}

Affiliations

¹ Department of Surgery, The First Affiliated Hospital of Naval Medical University Shanghai 200433, The People's Republic of China.
² Department of Hepatobiliary Pancreatic Surgery, Yueyang Hospital of Integrated Traditional Chinese and Western Medicine, Shanghai University of Traditional Chinese Medicine Shanghai 200437, The People's Republic of China.
³ Department of Pathology, Eastern Hepatobiliary Surgery Hospital, Third Affiliated Hospital of Naval Medical University Shanghai 200438, The People's Republic of China.
⁴ Shanghai GoBroad Cancer Hospital, China Pharmaceutical University Shanghai 200131, The People's Republic of China.
⁵ Department of Hepatic Surgery IV, The Eastern Hepatobiliary Surgery Hospital, Third Affiliated Hospital of Naval Medical University Shanghai 200438, The People's Republic of China.
⁶ Eastern Hepatobiliary Clinical Research Institute, Third Affiliated Hospital of Navy Medical University Shanghai 200438, The People's Republic of China.

Abstract

Background: The mechanism of ferroptosis is primarily driven by the iron-dependent lethal accumulation of membrane lipid peroxidation. Bavachin has been found to exacerbate lipid peroxidation in cancer cells; however, whether it hinders hepatocellular carcinoma (HCC) progression by way of ferroptosis remains unknown.

Methods: Cell counting kit-8 (CCK-8) assay was used to measure the effect of Bavachin on the viability of HCC cells, so as to determine the appropriate drug concentration for subsequent experiments. Combining molecular biology experimental techniques such as CCK-8, flow cytometry, western blotting, wound-healing assay, DCFH-DA (2',7'-dichlorodihydrofluorescein diacetate) fluorescent probe and a variety of biological kits, the effects of Bavachin on HCC cell malignant phenotype progression and ferroptosis were investigated.

Results: Bavachin significantly induced cytotoxicity of Huh-7 and HepG2 cells at concentrations of 20 and 40 μM, respectively. Bavachin intervention prominently reduced cell proliferation and migration, and enhanced cell apoptosis in HCC. Also, Bavachin enhanced the process of lipid peroxidation, as indicated by increased reactive oxygen species (ROS), lipid peroxidation (LPO) and malondialdehyde (MDA) production, and decreased superoxide dismutase (SOD) and glutathione (GSH) production. Ferrostin-1, a ferroptosis inhibitor, reduced the Bavachin-induced cell survival rate. Bavachin intervention induced ferroptosis by enhancing iron ion concentration, acyl-CoA synthetase long-chain family member 4 (ACSL4) expression, and reducing glutathione peroxidase-4 (GPX4) expression. Bavachin exerted anti-cancer effects though inducing ferroptosis by activating the nuclear factor erythroid 2-related factor 2 (Nrf2)/Heme oxygenase-1 (HO-1) pathway.

Conclusion: Bavachin acted as a ferroptosis inducer, promoted ROS release, enhanced lipid peroxidation, and inhibited HCC cell malignant phenotype progression by modulating the Nrf2/HO-1 pathway.

Keywords: Bavachin; ferroptosis; hepatocellular carcinoma; lipid peroxidation.