title: Immune modulation and apoptosis induction: Two sides of antitumoural activity of a standardised herbal formulation of Withania somnifera.
abstract: Deregulated apoptosis and suppressed tumour reactive immunity render tumour cells to grow amok in the host body. Traditionally used botanicals may offer potential anticancer chemo-immunotherapeutic leads. We report in this study a chemically standardised herbal formulation (WSF) of Withania somnifera possessing anticancer and Th1 immune up-regulatory activities. WSF produced cytotoxicity in a panel of human cancer cell lines in vitro. The molecular mechanism of cell cytotoxicity, IC(50) 48h approximately 20mug/ml, was investigated in HL-60, where it induced apoptosis by activating both intrinsic and extrinsic signalling pathways. It induced early generation of reactive nitrogen and oxygen species (RNOS), thus producing oxidative stress mediated mitochondrial membrane potential (MMP) loss leading to the release of cytochrome c, the translocation of Bax to mitochondria and apoptosis-inducing factor to the nuclei. These events paralleled the activation of caspase-9, -3 and PARP cleavage. WSF also activated caspase-8 through enhanced expression of TNF-R1 and DR-4, suggesting also the involvement of extrinsic pathway of apoptosis. WSF at 150mg/kg, i.p., inhibited >50% tumour growth in the mouse tumour models. In tumour-bearing mice, WSF inhibited the expression of pStat-3, with a selective stimulation of Th1 immunity as evidenced by enhanced secretion of IFN-gamma and IL-2. In parallel, it enhanced the proliferation of CD4(+)/CD8(+) and NK cells along with an increased expression of CD40/CD40L/CD80. In addition, WSF also enhanced T cell activation in camptothecin treated tumour-bearing mice. WSF being safe when given orally up to 1500mg/kg to rats for 6 months may be found useful in the management of malignancy by targeting at multiple pathways.
authors.affiliation: Division of Pharmacology, Indian Institute of Integrative Medicine (Council of Scientific and Industrial Research), Canal Road, Jammu 180001, India.
authors.forename: Fayaz
authors.initials: F
authors.lastname: Malik
authors.forename: Ajay
authors.initials: A
authors.lastname: Kumar
authors.forename: Shashi
authors.initials: S
authors.lastname: Bhushan
authors.forename: Dilip M
authors.initials: DM
authors.lastname: Mondhe
authors.forename: Harish C
authors.initials: HC
authors.lastname: Pal
authors.forename: Rohit
authors.initials: R
authors.lastname: Sharma
authors.forename: Anamika
authors.initials: A
authors.lastname: Khajuria
authors.forename: Surjeet
authors.initials: S
authors.lastname: Singh
authors.forename: Gurdarshan
authors.initials: G
authors.lastname: Singh
authors.forename: Ajit K
authors.initials: AK
authors.lastname: Saxena
authors.forename: Krishan A
authors.initials: KA
authors.lastname: Suri
authors.forename: Ghulam N
authors.initials: GN
authors.lastname: Qazi
authors.forename: Jaswant
authors.initials: J
authors.lastname: Singh
mesh_terms: D000818:Animals; D000972:Antineoplastic Agents, Phytogenic / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D003601:Cytotoxicity Tests, Immunologic; D004354:Drug Screening Assays, Antitumor; D018922:HL-60 Cells; D006801:Humans; D007166:Immunosuppressive Agents / Q000494:pharmacology*; D051379:Mice; D008807:Mice, Inbred BALB C; D008815:Mice, Inbred Strains; D009369:Neoplasms / Q000276:immunology* / Q000378:metabolism; D009569:Nitric Oxide / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism; D013601:T-Lymphocytes / Q000276:immunology; D032341:Withania
chemical_list: D000972:Antineoplastic Agents, Phytogenic; D007166:Immunosuppressive Agents; D017382:Reactive Oxygen Species; D009569:Nitric Oxide