Objectives: Artemisinins are antimalarial drugs that exert potent anticancer activity. We evaluated the effects of artesunate, a semisynthetic derivative of artemisinin, on tumor growth, angiogenesis, the unfolded protein response, and chemoresistance in hepatocellular carcinoma.
Materials and methods: The effect of artesunate was examined in HepG2 and BWTG3 cells under normoxic and hypoxic conditions and in a diethylnitrosamine-induced mouse model. Histology was performed with hematoxylin/eosin and reticulin staining. The expression of chemoresistance-related transporters and angiogenic and unfolded protein response factors was determined. Cytotoxicity was assessed by alanine and aspartate transaminase, lactate dehydrogenase, water-soluble tetrazolium salt, and caspase-3 activity assays. Small animal imaging was performed using dynamic contrast-enhanced MRI and choline PET to assess tumor progression.
Results: Artesunate dose dependently reduced cell viability (from 50 μmol/l; P<0.05) and increased caspase-3 activity (P<0.05) in HepG2 and BWTG3 cells. These effects were enhanced by hypoxia (from 12.5 μmol/l; P<0.01). Moreover, artesunate downregulated vascular endothelial growth factor and placental growth factor expression in vitro (both P<0.05) and in vivo (both P<0.01). In mice, artesunate decreased vessel density and tumor burden (both P<0.05). These in-vivo effects were enhanced by combination with sorafenib (P<0.05 and P=0.07, respectively), without apparent hepatotoxicity. Furthermore, artesunate modulated the unfolded protein response in vitro and in vivo, increasing proapoptotic signaling, and did not induce doxorubicin chemoresistance.
Conclusion: These findings indicate that artesunate could offer a new approach to the therapy of hepatocellular carcinoma. Clinical trials with artesunate as monotherapy or in combination with current hypoxia-inducing approaches are necessary.