Cantharidin Suppresses Cell Viability and Induces Apoptosis of SK-N-SH and SH-SY5Y Cells.

Abstract

Background/aim: Neuroblastoma (NBL) is a pediatric malignancy with high mortality, particularly within the first year of life. Cantharidin, a natural terpenoid derived from blister beetles, has shown anticancer activity against several malignancies; however, its effect on NBL remains unexplored. In this study, we evaluated the antiproliferative and pro-apoptotic effects of cantharidin on SH-SY5Y and SK-N-SH NBL cell lines.

Materials and methods: The cell viability and appearance of sub-G₁ were measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and flow cytometry, respectively. The alterations of apoptosis-related molecules were determined by western blotting.

Results: MTT assays revealed that treatment with 5 and 10 μM cantharidin for 24 and 48 h significantly reduced viability of both SH-SY5Y and SK-N-SH cells (all p<0.05), with 10 μM for 48 h reducing viability by 65.3% and 72.3%, respectively. Flow cytometry showed that 10 μM cantharidin induced apoptosis of 51.0% of SH-SY5Y and 68.3% of SK-N-SH cells at 48 h (p<0.05). Western blot analysis demonstrated increased expression of cleaved caspase-3, -8 and -9, and pro-apoptotic proteins BCL2-associated X (BAX) and BH3-interacting domain death agonist (BID), alongside reduced levels of anti-apoptotic BCL2 apoptosis regulator (BCL2) and B-cell lymphoma-extra large (BCL-xL). Cytochrome c release was also elevated, confirming mitochondrial pathway involvement. Additionally, phosphorylation of Janus kinase 2 (JAK2) and signal transducer and activator of transcription 3 (STAT3) was significantly suppressed by 10 μM cantharidin at 48 h, suggesting JAK2 and STAT3 pathway inhibition contributes to apoptosis.

Conclusion: These findings support the hypothesis that cantharidin induces apoptosis via both intrinsic and extrinsic pathways, as well as through suppression of the JAK2-STAT3 axis. Our results reveal that cantharidin holds significant promise as a multi-target therapeutic candidate for NBL, justifying additional in vivo validation and clinical translation efforts.