Immune modulation and apoptosis induction: Two sides of antitumoural activity of a standardised herbal formulation of Withania somnifera
Authors: Harish C Pal
Rohit Sharma
Krishan A Suri
Ajay Kumar
Shashi Bhushan
Fayaz Malik
Dilip M Mondhe
Jaswant Singh
Ajit K Saxena
Surjeet Singh
Anamika Khajuria
Ghulam N Qazi
Gurdarshan Singh
Journal: European Journal of Cancer (2009)
Discipline: CLINICAL MEDICINE
Keywords: Apoptosis; Immunity; Herbal formulation; Withania somnifera; Anticancer
Affiliations: Division of Pharmacology, Indian Institute of Integrative Medicine (Council of Scientific and Industrial Research), Canal Road, Jammu 180001, India.

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 &gt;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. 

Retracted: No

PubPeer comments:
 Could authors explain how they got such strong actin signals in nuclear fraction? What is the reliability of such data? ![file] , 
 The fact is that the presence of actin is not restricted to cytoplasmic only, nuclear actin is associated with numerous cellular functions and the same has been clearly shown in a number of reports publications available in the literature. Moreover, pertaining to this case, the cytoplasmic and nuclear fractions were prepared as per the indicated method (referred in the Methods section) and results are shown as per the experimental outcome. It is also possible that the fractionation protocol used was unable to separate 100% pure cytoplasmic nuclear fractions leading to enhanced signal. Despite that, this particular experimental outcome will not change the final conclusion of the paper. ,