Licochalcone A Induces Mitochondria-Dependent Apoptosis in Hepatocellular Carcinoma Cells via ROS-Regulated Activation of the p38/JNK/MAPK Pathway

Abstract

Background: Licochalcone A (LCA) has multiple pharmacological activities; however, its mechanism of action against hepatocellular carcinoma (HCC) remains incompletely defined. Objectives: This study aimed to investigate the inhibitory effects of LCA on HCC cells and to elucidate the mechanism by which LCA induces mitochondria-dependent apoptosis via a reactive oxygen species (ROS)-regulated p38/JNK pathway. Methods: HepG2 and HuH-7 HCC cell lines were used to establish an in vitro model and investigate the biological effects of LCA on HCC. Cell viability, injury, proliferative capacity, migration, and invasion were assessed to systematically evaluate the effects of LCA on the malignant phenotype of HCC cells. To explore the potential mechanisms by which LCA induces HCC cell injury, apoptosis, ROS levels, oxidative stress indicators, and mitochondria-related apoptotic indicators were measured. In mechanistic experiments, the ROS/p38/JNK/MAPK pathway was inhibited using the corresponding tool compounds, N-acetylcysteine, SB203580, and SP600125, to verify its role in mediating the proapoptotic effect of LCA. Results: LCA dose-dependently inhibited the viability, proliferation, migration, and invasion of HCC cells (P < 0.01) and induced apoptosis, without significant toxicity to normal THLE-2 hepatocytes. Licochalcone A significantly increased ROS accumulation in HCC cells, resulting in glutathione depletion, decreased superoxide dismutase activity, and elevated malondialdehyde levels (P < 0.01). Concurrently, LCA induced mitochondrial dysfunction, as evidenced by decreased mitochondrial membrane potential, reduced adenosine triphosphate production, calcium ion overload, and cytochrome c release (P < 0.01). Licochalcone A treatment activated the p38/JNK/MAPK signaling pathway, upregulated the expression of proapoptotic proteins, and downregulated the expression of antiapoptotic proteins (P < 0.01). These findings were confirmed by inhibitor experiments: N-acetylcysteine effectively suppressed LCA-induced ROS accumulation and pathway activation, whereas the combination of SB203580 and SP600125 partially attenuated LCA-mediated mitochondrial dysfunction, changes in apoptotic protein expression, and apoptosis. Conclusions: LCA exerts anti-HCC effects by inducing ROS accumulation, activating the p38/JNK pathway, modulating the balance of Bcl-2 family protein expression, and triggering mitochondria-dependent apoptosis.