Cantharidin Suppresses Cell Viability and Induces Apoptosis of SK-N-SH and SH-SY5Y Cells
Jen-Sheng Pei1, Hsu-Tung Lee2,3, Chao-Chun Chen1
1Department of Pediatrics, Taoyuan General Hospital, Ministry of Health and Welfare, Taoyuan, Taiwan, R.O.C.
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.
Insights
Cantharidin effectively reduces neuroblastoma (NBL) cell viability and induces apoptosis through intrinsic and extrinsic pathways. This natural compound also inhibits the JAK2-STAT3 axis, showing promise as a multi-target NBL therapeutic.
Area of Science:
- Pediatric Oncology
- Natural Product Chemistry
- Molecular Biology
Background:
- Neuroblastoma (NBL) is a high-mortality pediatric cancer.
- Cantharidin, a natural terpenoid, has shown anticancer effects but its impact on NBL is unknown.
Purpose of the Study:
- To investigate the antiproliferative and pro-apoptotic effects of cantharidin on NBL cell lines.
- To explore the molecular mechanisms underlying cantharidin's action in NBL.
Main Methods:
- MTT assays to assess cell viability.
- Flow cytometry to detect apoptosis.
- Western blotting to analyze apoptosis-related proteins and signaling pathways (JAK2/STAT3).
Main Results:
- Cantharidin significantly reduced viability and induced apoptosis in SH-SY5Y and SK-N-SH NBL cells.
- Apoptosis involved the mitochondrial pathway (cytochrome c release) and caspase activation.
- Cantharidin suppressed JAK2-STAT3 signaling and modulated BCL2 family proteins.
Conclusions:
- Cantharidin induces NBL cell apoptosis via intrinsic and extrinsic pathways.
- Inhibition of the JAK2-STAT3 axis contributes to cantharidin's apoptotic effect.
- Cantharidin is a promising multi-target therapeutic candidate for NBL, warranting further investigation.
