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Background <p><i>Withania somnifera</i> (L.) Dunal, commonly known as ashwagandha, is a cornerstone of Ayurvedic medicine and has demonstrated anti-metastatic properties, including the ability to mitigate the cytotoxic effects of carcinogens and chemotherapeutic agents. Neuroblastoma (NB), a highly aggressive paediatric cancer, accounts for approximately 15% of childhood cancer-related deaths. Despite intensive treatment, over 50% of NB cases experience tumor recurrence and debilitating long-term effects. This study aimed to evaluate the anti-progression effects of <i>W. somnifera</i> root fractions on the human NB Kelly cell line at sub-cytotoxic concentrations and to identify the active bioactive constituents.</p> Methods <p><i>W. somnifera</i> roots were extracted using 95% ethanol and subsequently fractionated <i>via</i> vacuum liquid chromatography with a methanol-water gradient elution, yielding twelve fractions. Kelly cells were treated with each fraction at sub-cytotoxic concentrations, as determined by MTT assay. Treated cells were then subjected to transwell extracellular matrix invasion and fibronectin adhesion assays. Data were analysed using one-way ANOVA (GraphPad Prism), with statistical significance set at <i>P</i> ≤ 0.05. Bioactive fractions were further subfractionated by preparative HPLC, and major constituents were tentatively identified using GC-MS.</p> Results <p>Fraction 9 (eluted with 70% methanol) exhibited the highest anti-invasive activity, whereas Fraction 10 (eluted with 80% methanol) demonstrated the most potent and statistically significant (<i>P</i> = 0.0409) anti-adhesive effect compared to vehicle-treated cells (0.5% DMSO). Subfraction analysis revealed that Subfraction 10/1 had a significant anti-adhesive effect (<i>P</i> = 0.0482), while subfractions 10/3 and 9/2 showed non-significant anti-adhesive effects. GC-MS analysis of subfractions 9/2, 10/1, and 10/3 revealed the presence of four previously unreported compounds in <i>W. somnifera</i>.</p> Conclusions <p>Constituents of <i>W. somnifera</i> roots exhibit promising anti-metastatic activity against neuroblastoma cells, highlighting their potential to complement existing chemotherapeutic regimens and reduce associated long-term side effects.</p> Clinical trial number <p>Not applicable.</p>

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Bioactivity-guided fractionation of Withania somnifera (L.) Dunal roots extract: evaluation of the anti-progressive potential on human Kelly neuroblastoma cell line

  • 期刊论文

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  • 发表日期：

    2025年7月16日

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  • Nada Ahmad Al-Hasawi,
  • Naser Faisal Al-Tannak,
  • Josily Joy,
  • Khaled Youssef Orabi

摘要

Background

Withania somnifera (L.) Dunal, commonly known as ashwagandha, is a cornerstone of Ayurvedic medicine and has demonstrated anti-metastatic properties, including the ability to mitigate the cytotoxic effects of carcinogens and chemotherapeutic agents. Neuroblastoma (NB), a highly aggressive paediatric cancer, accounts for approximately 15% of childhood cancer-related deaths. Despite intensive treatment, over 50% of NB cases experience tumor recurrence and debilitating long-term effects. This study aimed to evaluate the anti-progression effects of W. somnifera root fractions on the human NB Kelly cell line at sub-cytotoxic concentrations and to identify the active bioactive constituents.

Methods

W. somnifera roots were extracted using 95% ethanol and subsequently fractionated via vacuum liquid chromatography with a methanol-water gradient elution, yielding twelve fractions. Kelly cells were treated with each fraction at sub-cytotoxic concentrations, as determined by MTT assay. Treated cells were then subjected to transwell extracellular matrix invasion and fibronectin adhesion assays. Data were analysed using one-way ANOVA (GraphPad Prism), with statistical significance set at P ≤ 0.05. Bioactive fractions were further subfractionated by preparative HPLC, and major constituents were tentatively identified using GC-MS.

Results

Fraction 9 (eluted with 70% methanol) exhibited the highest anti-invasive activity, whereas Fraction 10 (eluted with 80% methanol) demonstrated the most potent and statistically significant (P = 0.0409) anti-adhesive effect compared to vehicle-treated cells (0.5% DMSO). Subfraction analysis revealed that Subfraction 10/1 had a significant anti-adhesive effect (P = 0.0482), while subfractions 10/3 and 9/2 showed non-significant anti-adhesive effects. GC-MS analysis of subfractions 9/2, 10/1, and 10/3 revealed the presence of four previously unreported compounds in W. somnifera.

Conclusions

Constituents of W. somnifera roots exhibit promising anti-metastatic activity against neuroblastoma cells, highlighting their potential to complement existing chemotherapeutic regimens and reduce associated long-term side effects.

Clinical trial number

Not applicable.