Selective Cytotoxic Effects of Amygdalin on Neuroblastoma Cells: Mechanisms of Apoptosis, Autophagy and Anti-Metastatic Potential

Abstract

Objectives: Neuroblastoma is the most common nervous system cancer in children. Amygdalin, a glycoside with known anticancer properties found in fruit seeds, has not been previously studied in the context of neuroblastoma. This study investigated amygdalin’s cytotoxic and anti-migratory effects on SH-SY5Y and L929 cells via cellular mechanisms. Materials and Methods: In this study, cells were treated with amygdalin (0.5–50 mM), then cell viability was measured by the MTS method and cell migration by the wound healing assay. Gene expressions associated with autophagy and apoptotic cell death (Bax, Bcl-2, Caspase-3 (Cas-3), Cas-8, PI3K, Akt, mTOR, SQSTM-1, and Beclin-1) were measured using RT-PCR. Results: Amygdalin exhibited selective cytotoxicity, with a more pronounced effect on SH-SY5Y neuroblastoma cells compared to L929 fibroblasts, as indicated by lower IC50 values (3.27 mM for SH-SY5Y; 7.24 mM for L929). Amygdalin significantly inhibited migration in SH-SY5Y cells, significantly increasing wound area (26.36±1.98% at 24h to 62.65±9.16% at 48h), but had a limited effect on L929 cells (20.32±1.73% to 19.79±1.40% h). In L929 cells, amygdalin partially inhibited apoptosis by upregulating Cas-8 and Bcl-2, downregulating Cas-3 and Bax, and also suppressed autophagy by reducing Beclin-1 expression. In contrast, in SH-SY5Y cells, amygdalin activated survival signaling via the PI3K/Akt/mTOR pathway while simultaneously promoting apoptosis (increased expression of Bax, Cas-3, and Cas-8) and autophagy (increased levels of Beclin-1). Conclusion: Although amygdalin activates survival pathways in SH-SY5Y cells, increased stress may override these signals, leading to apoptotic cell death, suggesting a potential therapeutic role in neuroblastoma treatment.

Keywords

• Amygdalin
• Neuroblastoma
• Apoptosis
• Autophagy
• Cytotoxic

Supporting Institution

This study was supported by the TÜBİTAK 2209-A University Students Research Projects Support Program (project number 1919B012325433).

Project Number

Project number 1919B012325433

Ethical Statement

This study was performed using continuous (immortalized) cell lines and does not include any direct data or samples from human or animal subjects

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