Investigation of the anti neoplastic activity of Vitex agnus castus L. essential oil on neuroblastoma cells

ABSTRACT INVESTİGATİON OF THE ANTİ NEOPLASTİC ACTİVİTY OF VİTEX AGNUS CASTUS L. ESSENTİAL OİL ON NEUROBLASTOMA CELLS Aim: The aim of this study is to evaluate the potential antineoplastic (anti-tumor) effects of the essential oil obtained from Vitex agnus-castus L., a plant known in traditional medicine for its various pharmacological properties, on neuroblastoma cells—one of the most common childhood cancers—under in vitro conditions. In this context, the effects of the essential oil on cell proliferation, apoptosis induction, and cytotoxicity are intended to be determined. Material and Method: In this study, the antineoplastic effects of the essential oil obtained from Vitex agnus-castus L. through traditional distillation were investigated in vitro on the human neuroblastoma cell line (SH-SY5Y). The cells were cultured in DMEM medium and treated with various concentrations of the essential oil. The cytotoxic effects were assessed using the XTT cell viability assay, and the IC₅₀ value was calculated. Apoptosis levels were determined using the TUNEL method, while cell migration was evaluated via the wound healing assay. Additionally, RT-PCR analyses were conducted to examine the effect of VAC treatment on the expression levels of specific genes. All experimental data were statistically analyzed, and a significance level of p<0.05 was considered. Results: Vitex agnus-castus L. essential oil exhibited a dose-dependent cytotoxic effect on SH-SY5Y neuroblastoma cells, significantly reducing cell viability. While the expression of apoptotic genes such as NOXA, CASP3, and DR5 was downregulated, an increase was observed in BDNF, MAP2, and NLGN3 gene expression. Although TUNEL analysis showed a slight increase in apoptosis, this change was not statistically significant. However, wound healing assays revealed a marked inhibition of cell migration. These findings suggest that VAC may possess differentiation-promoting and anti-metastatic potential rather than classical pro-apoptotic activity. Conclusions: Our study suggests that Vitex agnus castus essential oil could be a potential natural agent in neuroblastoma treatment, and further research is needed to comprehensively elucidate its mechanisms of action, investigate other cell death pathways, identify its bioactive compounds, and validate these findings through in vivo studies. Keywords: Antineoplasticity, Apoptosis, Cytotoxicity, Neuroblastoma, SH-SY5Y, Vitex Agnus Castus L.