introduction: Artesunate exhibits potent anti-cancer effects, but its mechanism of action in lenvatinib resistant of hepatocellular carcinoma (HCC) remains unclear. This study aimed to investigate the effects of artesunate on lenvatinib efficacy and ferroptosis in HCC and to elucidate the underlying mechanisms. materials and methods: RNA sequencing (RNA-seq) was used to screen for the differentially expressed gene- solute carrier family 25 member 27 (SLC25A27) in lenvatinib-resistant (Lenva-R) HCC cell lines. Cell viability and proliferation were assessed using CCK-8, clone formation assay, and apoptosis assay. Western blot, qRT-PCR, and immunohistochemistry were used to verify SLC25A27 and ferroptosis-related protein expression. GSEA was used to analyses the functional enrichment pathway of SLC25A27. Transmission electron microscopy was used to observe the subcellular localization and the status of mitochondrial ferroptosis. Ferroptosis metabolism was evaluated by measuring intracellular Fe2+, ROS, MDA, and GSH/GSSG assay. Gain-of-function and loss-of-function experiments were performed to elucidate the biological function of SLC25A27 in HCC. The effect of signal transducer and activator of transcription 3 (STAT3) on SLC25A27 transcription was investigated using dual-luciferase reporter assay. The effect of artesunate on lenvatinib sensitivity was examined through in vivo and in vitro experiments. results: The expression of SLC25A27 was upregulated in Lenva-R HCC cells and tissues. In vitro experiments confirmed that SLC25A27 promoted HCC cell proliferation, and reduced apoptosis, as well as inhibiting ferroptosis. Mechanistically, SLC25A27 was negatively associated with functions of oxidative stress, which could inhibit ferroptosis and lead to lenvatinib resistance. Dual-luciferase reporter experiments confirmed that STAT3 significantly enhances SLC25A27 transcription. discussion: Artesunate inhibited STAT3/SLC25A27/ glutathione peroxidase 4 (GPX4) expression and promoted ferroptosis in vivo and in vitro, significantly enhancing the anti-HCC effect of lenvatinib. conclusion: Artesunate ameliorates lenvatinib resistance by inhibiting the STAT3/SLC25A27 pathway and promoting ferroptosis in HCC.
Bi et al. (Tue,) studied this question.