Withania somnifera Water Extract as a Potential Candidate for Differentiation Based Therapy of Human Neuroblastomas

Abstract

Neuroblastoma is an aggressive childhood disease of the sympathetic nervous system. Treatments are often ineffective and have serious side effects. Conventional therapy of neuroblastoma includes the differentiation agents. Unlike chemo-radiotherapy, differentiation therapy shows minimal side effects on normal cells, because normal non-malignant cells are already differentiated. Keeping in view the limited toxicity of Withania somnifera (Ashwagandha), the current study was aimed to investigate the efficacy of Ashwagandha water extract (ASH-WEX) for anti-proliferative potential in neuroblastoma and its underlying signalling mechanisms. ASH-WEX significantly reduced cell proliferation and induced cell differentiation as indicated by morphological changes and NF200 expression in human IMR-32 neuroblastoma cells. The induction of differentiation was accompanied by HSP70 and mortalin induction as well as pancytoplasmic translocation of the mortalin in ASH-WEX treated cells. Furthermore, the ASH-WEX treatment lead to induction of neural cell adhesion molecule (NCAM) expression and reduction in its polysialylation, thus elucidating its anti-migratory potential, which was also supported by downregulation of MMP 2 and 9 activity. ASH-WEX treatment led to cell cycle arrest at G0/G1 phase and increase in early apoptotic population. Modulation of cell cycle marker Cyclin D1, anti-apoptotic marker bcl-xl and Akt-P provide evidence that ASH-WEX may prove to be a promising phytotherapeutic intervention in neuroblatoma related malignancies.

Withania somnifera Suppresses Tumor Growth of Intracranial Allograft of Glioma Cells

“…Overexpression of the cyclin D1 gene has been reported in several human neoplasms [50,51], and elevated expression of cyclin D1 is associated with high degree of malignancy and rapid cell proliferation [52]. Arrest of the cell cycle at the G2/M phase in vitro is one of the effects of ASH-WEX found in this study, which is in agreement with our previous study with neuroblastoma and other cell lines [19,53] and other ashwagandha extracts [15]. Annexin V-FITC/PI staining study further supports this observation as there is an increase in early apoptotic cell population which may be due to differentiation-inducing ability of ASH-WEX which may finally leading the treated cells to normal apoptotic pathway.…”

Biotechnological interventions inWithania somnifera(L.) Dunal

“…Majority of these tumors develop into malignancy and remain incurable in spite of the therapies like external beam radiation, surgery, and chemotherapy and hence, call for the development of novel therapeutic approaches. W. somnifera induced upregulation of neuronal marker proteins has been correlated with the induction Biotechnology and Genetic Engineering Reviews 11 of differentiation in these cells (Kataria et al, 2011(Kataria et al, , 2013Kuboyama et al, 2002;Shah et al, 2009). The upregulation of glial fibrillary acidic protein (GFAP) in glioma, neurofilament protein (NF200) in neuroblastoma, and mortalin expression suggested the induction of differentiation in these cells.…”

“…The upregulation of glial fibrillary acidic protein (GFAP) in glioma, neurofilament protein (NF200) in neuroblastoma, and mortalin expression suggested the induction of differentiation in these cells. The upregulation of neural cell adhesion molecule (NCAM) and downregulation of its polysialylated form (PSA-NCAM) and matrix metalloproteases (MMPs) may explain the anti-migratory and differentiation inducing properties of W. somnifera leave extract in the glioblastoma and neuroblastoma cells (Kataria et al, 2011(Kataria et al, , 2013. Further, W. somnifera treatment led to cell cycle arrest at G0/G1 phase and increase in early apoptotic population along with modulation of cell cycle marker Cyclin D1, anti-apoptotic marker bcl-xl, and Akt-P (Kataria et al, 2013;Widodo et al, 2007).…”

“…The upregulation of neural cell adhesion molecule (NCAM) and downregulation of its polysialylated form (PSA-NCAM) and matrix metalloproteases (MMPs) may explain the anti-migratory and differentiation inducing properties of W. somnifera leave extract in the glioblastoma and neuroblastoma cells (Kataria et al, 2011(Kataria et al, , 2013. Further, W. somnifera treatment led to cell cycle arrest at G0/G1 phase and increase in early apoptotic population along with modulation of cell cycle marker Cyclin D1, anti-apoptotic marker bcl-xl, and Akt-P (Kataria et al, 2013;Widodo et al, 2007). These studies provide evidence that W. somnifera appears to affect multiple pathways for its anti-cancer and differentiation-inducing role in glioma and neuroblastoma cells instead of targeting a single protein or pathway.…”

Identification of amino acid appended acridines as potential leads to anti-cancer drugs

“…15 These observations are also supported by previous Letters on anticancer and differentiation inducing activities of some natural products. [16][17][18] This upregulated expression of mortalin (Figs. 4A, 5B and C) was also followed by elevated expression of mitochondrial stress response protein HSP70.…”