Artesunate, a drug best known for its lifesaving role in malaria treatment, may have a second life as a targeted therapy against hepatocellular carcinoma, the most common primary cancer of the liver. A new study reports that artesunate directly binds to and inhibits glucosylceramidase, or GBA, an enzyme involved in sphingolipid metabolism. By disrupting this metabolic process, the drug triggered a chain of molecular events that damaged mitochondria and activated programmed cell death in liver cancer cells. The findings provide a structural explanation for artesunate’s anticancer activity and identify a previously underexplored therapeutic vulnerability in hepatocellular carcinoma.
The research, conducted by scientists from the China Academy of Chinese Medical Sciences and Fujian University of Traditional Chinese Medicine, addresses a major challenge in liver cancer treatment. Hepatocellular carcinoma often develops in the context of chronic liver disease and can be difficult to control once it has progressed. Although surgery, ablation, immunotherapy, targeted drugs, and chemotherapy can benefit selected patients, treatment resistance and disease recurrence remain widespread. Artesunate is already widely used against malaria, giving it an established pharmacological history and a well-characterized clinical profile. However, the molecular basis of its activity against cancer has remained incompletely understood.
The investigators first examined how artesunate affected the survival and growth of two human hepatocellular carcinoma cell lines, HepG2 and MHCC-97H. Using the CCK8 assay, which measures cellular metabolic activity as an indicator of viability and proliferation, they found that artesunate inhibited both cell lines in a concentration-dependent manner. HepG2 cells were more sensitive than MHCC-97H cells, suggesting that differences in metabolic state or drug-response pathways may influence the treatment’s effectiveness. Additional experiments showed that artesunate reduced cancer-cell proliferation and increased the proportion of cells undergoing apoptosis, a tightly regulated form of cell death that is frequently disabled in tumors.
The study also examined artesunate in an orthotopic mouse model, in which HepG2 cells were injected into the liver to reproduce a more realistic tumor environment than conventional subcutaneous models. Animals receiving low, middle, or high doses of artesunate showed evidence of increased tumor-cell apoptosis. TUNEL staining, which detects fragmented DNA associated with programmed cell death, and Hoechst staining, which reveals changes in nuclear structure, both supported the conclusion that artesunate promoted apoptosis in the tumors. The effects were compared with control animals and with a group receiving 5-fluorouracil, a commonly used anticancer drug. These experiments provided in vivo support for the cellular findings, although additional animal and clinical studies will be necessary to determine whether the effect can be translated into a useful treatment.
The researchers connected artesunate’s activity to sphingolipid metabolism, a biochemical network that produces and regulates lipids involved in membrane structure, cell signaling, inflammation, and cell death. GBA normally helps break down glucosylceramide, a glycosphingolipid, into downstream metabolic products. When GBA was inhibited by artesunate, glucosylceramide-related metabolites accumulated and the balance of cellular sphingolipids was disturbed. Such metabolic changes can place stress on organelles and alter signaling pathways that control survival. In the treated liver cancer cells, this disruption was associated with mitochondrial dysfunction, a critical event because mitochondria regulate the intrinsic pathway of apoptosis.
The study describes a signaling sequence linking altered lipid metabolism to mitochondrial apoptosis: GBA, ceramide, cathepsin D, alpha-synuclein, BID, and BAX. In this proposed GBA–ceramide–CTSD–α-syn–BID–BAX axis, artesunate first suppresses GBA activity, altering ceramide metabolism. The resulting biochemical imbalance interferes with the maturation or function of cathepsin D, a lysosomal protease. It also promotes the accumulation of alpha-synuclein, a protein better known for its association with neurodegenerative disease but increasingly recognized as a regulator of cellular stress and organelle communication. These changes facilitate cleavage of BID and increase the activity or abundance of BAX, two important components of the mitochondrial death pathway. BAX can promote mitochondrial membrane permeabilization, allowing apoptotic factors to escape and activate downstream caspases, the enzymes that dismantle the cell.
Rescue experiments strengthened the proposed mechanism. When researchers supplemented cells with ceramide, they were able to influence the apoptotic response, supporting the idea that sphingolipid imbalance lies between GBA inhibition and mitochondrial damage. Conversely, suppressing alpha-synuclein reduced key effects of artesunate, indicating that alpha-synuclein accumulation is not simply a passive consequence of treatment but contributes to the death signal. The researchers also tested LTI-291, described in the study as a GBA enzyme activator, in combination with high-dose artesunate. The combined treatment helped probe whether restoring GBA-related activity could counteract artesunate’s effects. Together, these interventions provided functional evidence that the pathway is central to the drug’s anticancer action rather than being an incidental molecular signature.
A particularly significant part of the work focused on the physical interaction between artesunate and GBA. Through computational modeling and biochemical analyses, the researchers identified three amino-acid residues—tyrosine 313, glutamate 340, and asparagine 396—as important potential contact points within the enzyme’s active site. Site-directed mutagenesis was then used to replace selected residues and test their importance experimentally. Mutations affecting E340 and N396 substantially weakened artesunate binding and reduced GBA enzymatic activity. The altered enzyme also lost much of its ability to transmit the downstream apoptotic response induced by artesunate. These results support a direct target-engagement model in which the drug’s chemical structure fits into a functional region of GBA and changes the enzyme’s behavior.
The findings are notable because they move beyond the observation that artesunate can kill cancer cells and begin to explain why. Drug repurposing often starts with a promising biological effect, but successful development requires knowledge of the target, the binding site, the responsive cancer subtypes, and the mechanisms that may produce resistance. By defining GBA as a direct molecular target and connecting it to a lipid-regulated apoptotic pathway, the study offers several possible directions for future research. GBA expression or sphingolipid profiles might eventually help identify tumors most likely to respond, while combinations involving ceramide metabolism, lysosomal function, or mitochondrial apoptosis could potentially improve treatment activity.
At the same time, the results should not be interpreted as evidence that artesunate is already an established liver cancer therapy. The experiments were performed mainly in cultured cell lines and mouse models, systems that cannot fully reproduce the genetic diversity, immune environment, drug metabolism, and treatment history of human tumors. The greater sensitivity of HepG2 cells compared with MHCC-97H cells also highlights the possibility that response depends on tumor-specific biology. Future work will need to test the mechanism in patient-derived organoids, genetically diverse xenograft models, and carefully designed pharmacological studies. Long-term safety, optimal dosing, interactions with current liver cancer treatments, and the effects of artesunate on healthy liver tissue will also require detailed evaluation.
Published in Genes & Diseases, the study presents artesunate as more than an antimalarial compound with broad anticancer activity. It identifies a defined enzyme target, maps critical binding residues, and traces a mechanistic route from altered sphingolipid metabolism to mitochondrial apoptosis in hepatocellular carcinoma. If the findings are confirmed in clinically relevant models and human studies, GBA-targeted strategies could expand the therapeutic possibilities for a cancer that continues to demand more effective and durable treatments. For now, the work provides a compelling molecular blueprint for investigating how an established medicine might be redesigned or repurposed to exploit metabolic weaknesses in liver cancer.
Subject of Research: Artesunate-induced apoptosis and GBA-targeted mechanisms in hepatocellular carcinoma
Article Title: Artesunate directly targets glucosylceramidase to suppress hepatocellular carcinoma proliferation and trigger apoptosis
Web References: https://doi.org/10.1016/j.gendis.2026.102045; https://www.sciencedirect.com/journal/genes-and-diseases
References: Genes & Diseases, DOI: 10.1016/j.gendis.2026.102045
Image Credits: Xia Mao, Xiangying Yan, Yawen Chen, Bingbing Cai, Wenjia Chen, Ya Lin, Na Lin, Yanqiong Zhang
Keywords
Artesunate, hepatocellular carcinoma, liver cancer, glucosylceramidase, GBA, sphingolipid metabolism, ceramide, mitochondrial apoptosis, cathepsin D, alpha-synuclein, BID, BAX, drug repurposing
