Indian Drugs, 2009, 46(8): 603-609
ANTICANCER ACTIVITY OF WITHANIA SOMNIFERA DUNAL
Madhuri, Sharma1 and Pandey, Govind2*
1Senior Research Fellow of C.S.I.R. & Ph.D. Scholar, Department of Zoology & Biotechnology,
Model Science College, Jabalpur- 482001 (M.P.); 2Officer-In-Charge of Rinder Pest (Animal
Husbandry Department, Govt. of M.P.), Jabalpur
Considerable research works have been done on Withania somnifera (Ashwagandha)
to evaluate its anticancer activity, and some of the works have provided very promising
results. W. somnifera reduced the cancer cell proliferation and increased the overall
survival time. It enhanced the effectiveness of radiation therapy. It also reduced the side
effects of some chemotherapeutic agents, viz. cyclophosphamide and paclitaxel without
interfering with the cancer-reducing actions of the drugs. Given its broad spectrum of
cytotoxic and anticancer activity, W. somnifera presents itself as a novel therapy for
cancer. However, there is a great need to derive the potent anticancer drug from this
medicinal plant by conducting thorough research work.
Keywords: Withania somnifera, Withanolides, Anticancer activity
W. somnifera, also known as Ashwagandha, Indian ginseng, asgandh and winter cherry,
belongs to plant family Solanaceae. It grows in India, Pakistan, Afghanistan, Spain, parts of the
Middle-East Africa and Canary Islands. The plant is an erect undershrub that grows up to 1.5
meters tall. It has been an important herb in the Ayurvedic and indigenous medicine systems for
over 3000 years1-2. Various parts of the plant are traditionally used in the various diseases. Root
is traditionally used as aphrodisiac, liver tonic, antiinflammatory, astringent and antidepressant
and in impaired memory, neurasthenic, poor muscle tone and fever3. Western research supports
its polypharmaceutical use, confirming antioxidant, antiinflammatory, immunomodulating and
*Author for correspondence and reprint: Dr. Govind Pandey, 157/1, Badhaiyapura, North Milloniganj,
Jabalpur- 482002, M.P.; E-mail: firstname.lastname@example.org; email@example.com;
antistress properties in the whole plant extract and several separate constituents4. Anxiolytic and
antidepressant actions of the glycowithanolides, isolated from the roots of W. somnifera, in rats
were assessed5. W. somnifera inhibited stress-induced gastric ulcer more effectively as compared
to the standard drug ranitidine6. It significantly inhibited haloperidol or reserpine-induced
catalepsy and provided hope for the treatment of Parkinson's disease7. It also possessed
antivenom8 and antiinflammatory activities9. It also exhibited antiangiogenic effect10. Its root is a
potential source of hypoglycaemic, diuretic and hypocholesterolemic drugs11. Methanol and
hexane extracts of leaves and roots showed potent antibacterial activity12. Its cardioprotective
effect has also been reported experimentally13. In animals, W. somnifera acts as an
anticonvulsant in acute and chronic seizure models14. The glycoprotein of W. somnifera has
antifungal activity15. It also possesses adaptogenic, cardiotropic and anticoagulant properties16.
Leaf, root and root bark of W. somnifera possessed antimalarial activity17. Withaferin A, a major
chemical constituent of W. somnifera, possesses antiarthritic, antimicrobial, antimitotic and
Besides above properties, W. somnifera has also been reported to possess antitumour/
anticancer property. The present article explores out the research literatures pertaining to
anticancer activity of W. somnifera.
PHYTOCONSTITUENTS OF W. SOMNIFERA
The majority of the phytoconstituents of W. somnifera are withanolides (steroidal lactones
with ergostane skeleton) and alkaloids. These include withanone, withaferin A, and several other
withanolides and withasonidienone19. Much of the pharmacological activity of W. somnifera has
been attributed to two main withanolides, withaferin A and withanolide D20. Apart from these
contents, this plant also contains chemical constituents like withaniol, acylsteryl glucosides,
starch, reducing sugar, hantreacotane, ducitol, a variety of amino acids including aspartic acid,
proline, tyrosine, alanine, glycine, glutamic acid, cystine, tryptophan and high amount of iron3,21.
Most of these compounds have been found in both aqueous and alcoholic (ethanol) extracts of W.
somnifera root. The phytochemicals like alkaloids, reducing sugars, resins, saponins, fixed oils,
anthraquinones, proteins and amino acids have been present in the aqueous and alcoholic
extracts, while glycosides have been found in the alcoholic extract (AlE) of W. somnifera root22.
ANTICANCER ACTIVITY OF W. SOMNIFERA
The AlE of whole plant of W. somnifera (200 mg/kg, orally daily for seven months) reduced
the tumour incidence significantly against urethane (125 mg/kg biweekly for seven months)-
induced lung adenomas in adult male albino mice. The histological appearance of the lungs of
animals protected by W. somnifera was similar to those observed in the lungs of control
animals23. This drug has been found to scavenge the reactive molecules leading to
antimutagenesis and anticarcinogenesis24. The AlE of the root of W. somnifera was injected
intraperitonially (ip) at daily doses of 200 to 1000 mg/kg for 15 days starting from 24 hr after
intradermal inoculation of 5×10(5) cells of Sarcoma 180 (S 180) in Balb/c mice. Cumulative
doses of 7.5 to 10 g, ip at daily doses of 500 or 750 mg/kg seems to produce a good response in
this tumour25. The antitumour effect of root extract of W. somnifera and its modification by heat
were studied in vivo on S 180 tumour grown on the dorsum of adult Balb/c mouse26. The AlE of
the root of this plant produced in vivo growth inhibitory effect on transplantable mouse tumour, S
18027. Further, the AlE of the dried root of this plant as well as its active component withaferin A
showed significant antitumour effect in experimental tumours in vivo28. The author also observed
the anticancer effect of withaferin A against Ehrlich ascites carcinoma (EAC) in vivo29. In
another study, withaferin A was found to inhibit tumour growth and increased tumour free
survival in a dose-depended manner30.
W. somnifera may be used as an adjuvant during cancer chemotherapy for prevention of
bone marrow depression associated with anticancer drugs31. Administration of the root extract
(20 mg/dose/animal, ip) of W. somnifera was found to inhibit the 20-methylcholanthrene induced
sarcoma in mice and increase the life span of tumour bearing animals32. Administration of this
extract also reduced the skin carcinogenesis induced by dimethyl benzanthracene (DMBA) and
croton oil. W. somnifera treated animals showed increased glutathione (GSH), superoxide
dismutase (SOD), glutathione peroxidase (GPX) and catalase (CAT) in the liver and skin33.
Methanol extracts of W. somnifera root at a dose of 65 μg/ml or 265 μg/ml were able to down-
regulate the expression of p34cdc2, a cell-cycle regulatory protein. This protein is expressed
during cellular proliferation, and down regulation arrests the cell cycle in the G2/M transition
phase34. This plant showed a significant increase in cytotoxic T lymphocyte production both in
vivo as well as in vitro and it may reduce tumour growth35. W. somnifera (root extract) treated
splenocytes along with the mitogen lipopolysaccharide (LPS) could stimulate the lymphocyte
proliferation six times more than the normal. The natural killer (NK) cell activity was enhanced
significantly in both the normal and tumour bearing animals36. Oral treatment with
hydroalcoholic extract (HAE) of W. somnifera root at the dose of 400 mg/kg (one week before
injecting 20-methylcholanthrene and continued until 15 weeks thereafter) significantly reduced
the tumour incidence, tumour volume and enhanced the survival of the mice, compared with 20-
methylcholanthrene injected mice bearing with fibrosarcoma37.
The chemopreventive effect was demonstrated in a study of HAE of W. somnifera root on
DMBA induced skin cancer in mice. A significant decrease in incidence and average number of
skin lesions was noticed. Additionally, levels of reduced GSH, SOD, CAT and GPX in exposed
tissue returned to near normal values following administration of the extract. The
chemopreventive activity is thought to be due in part to antioxidant/ free radical scavenging
activity of the extract38. W. somnifera root possessed anticancer activity in swiss albino mice
bearing EAC and S 180 tumours22. Withaferin A from W. somnifera was most effective at
delaying tumour growth and doubling time in fibrosarcoma39. On in vitro study, withanolides
inhibited the growth in human breast (MCF-7), central nervous system (SF-268), lung (NCL-
H460) and colon (HCT-116) cancer cell lines comparable to doxorubicin. Withaferin A more
effectively inhibited the growth of breast and colon cancer cell lines than did doxorubicin. These
results suggest that the extracts of W. somnifera root may prevent or inhibit tumour growth in
cancer patients, and suggest a potential for development of new chemotherapeutic agents40. It has
been reported that AlE of W. somnifera is effective against different prostate cancer cell lines of
various metastatic potential41. Both in vivo and in vitro research attest to the cytotoxic and
antitumour potential of W. somnifera. The osteogenic sarcoma and breast carcinoma cell lines
were treated with 3-24 μg/ml aqueous leaf powder extract of W. somnifera. These cancers
exposed to high oxidative stress via a high-glucose medium or exposure to H2O2 were more
susceptible to oxidative damage after treatment with W. somnifera extract, suggesting that drug
has antiproliferative effect on tumour cells42.
A significant increase in the life span and a decrease in the cancer cell number and tumour
weight were noted in the tumour-induced mice after treatment with AlE of W. somnifera root.
The haematological parameters were also corrected. These observations are suggestive of the
protective effect of W. somnifera in Dalton's Ascitic Lymphoma43. Pretreatment with the animals
with 1-oxo-5 beta, 6 beta-epoxy-witha-2-enolide (20 mg/kg), isolated from the roots of W.
somnifera, prior to exposing the animals to ultraviolet (UV) radiation, prevents the incidence of
skin carcinoma44. Pretreatment of W. somnifera root powder extract (20 mg/dose/animal/24 hr, ip
for 10 days) or a constituent of W. somnifera, withanolide D (500 μg/dose/animal/24 hr, ip for 10
days) resulted in a significant reduction in tumour (melanoma) and increase in life span of
mice45. W. somnifera inhibited benzo(a)pyrene induced fore-stomuch papillomagenesis, showing
up to 60% and 92% inhibition in tumour incidence and multiplicity, respectively. Similarly, W.
somnifera inhibited the DMBA induced skin papillomagenesis, showing up to 45% and 71%
inhibition in tumour incidence and multiplicity46. The benzo(a)pyrene induced cancer animals
were treated with W. somnifera extract for 30 days, resulting into significant alteration of the
levels of immunocompetent cells, immune complexes and immunoglobulins47. It was also
observed that the combination chemotherapy of W. somnifera along with paclitaxel is a
promising chemotherapeutic agent against lung cancer induced by benzo(a)pyrene in swiss
albino mice48. The antiproliferative activity was screened against human laryngeal carcinoma
(Hep2) cells by microculture tetrazolium assay. The effect was confirmed in vivo by mouse
sponge implantation method. The experiments suggest that the roots of W. somnifera possess cell
cycle disruption and antiangiogenic activity, which may be a mediator for its anticancer action49.
Withanolides inhibited the cyclooxygenase enzymes, lipid peroxidation and proliferation of
tumour cells. It suppressed NF-kappaB (NF-kB) activation induced by a variety of inflammatory
and carcinogenic agents including tumour necrosis factor (TNF), interleukin-1beta, doxorubicin
and cigarette smoke condensate. Suppression was not cell type specific, as both inducible and
constitutive NF-kB activation were blocked by withanolides. The suppression occurred through
the inhibition of inhibitory subunit of I-kappaB (I-kB) alpha kinase activation, I-kB alpha
phosphorylation, I-kB alpha degradation, p65 phosphorylation and subsequent p65 nuclear
translocation. Overall, withanolides inhibit the activation of NF-kB and NF-kB-regulated gene
expression, which may explain the ability of withanolides to enhance apoptosis and inhibit
invasion and osteoclastogenesis50. The aqueous extract of W. somnifera had inhibitory effect on
Chinese hamster ovary cell lines51. Withaferin A induced the prostate apoptosis responses-4 in
androgen-refractory prostate cancer cells, showing that W. somnifera exhibits the cytotoxic effect
against variety of cancer cell lines52. W. somnifera primarily induced the oxidative stress in
human leukemia HL60 cell and in several other cancer cell lines53. It was further pointed that the
withaferin A inhibited DNA binding of NF-kB and caused nuclear cleavage of P65/Rel by
activated caspase-3. N acetyl-cysteine rescued all these events suggesting thereby a pro-oxidant
effect of Withaferin A. Apart from these, the anticancer activity of W. somnifera has also been
mentioned by several other workers20,54-62.
POSSIBLE ANTICANCER MECHANISM OF W. SOMNIFERA
The anticancer activity of W. somnifera is related to its multiple functions. W. somnifera
may increase the overall effectiveness of cancer treatment. Its anticancer activity is probably due
to the action of its main constituents, viz. withaferin A (which inhibits RNA and protein
production) and withanolide D (which inhibits RNA production). The RNA and protein
inhibition may lead to increased cancer cell death1. W. somnifera exhibits both antioxidant and
pro-oxidant activities. Tumour-bearing animals treated with this drug showed increased GSH,
SOD, GPX and CAT in the liver and skin33,46. These effects could clearly repair oxidative
damage caused by tumour growth and inflammation, thus reducing the likelihood of disease
progression. This antioxidant activity is enhanced by the potential of W. somnifera to up-regulate
phase II liver enzymes. W. somnifera may also mitigate unregulated cell growth via the potent
tumour suppressor gene p53, which regulates cell cycle proliferation44. W. somnifera was
identified via mass spectrometry as the most potent constituent of W. somnifera to inhibit tumour
TNF-α induced NF-kB activation, inhibiting angiogenesis at a dose of 7 μg/kg/day. The NF-kB
may play a key role in the antitumour action of W. somnifera since it is activated by carcinogens,
tumour promoters and inflammatory agents. This implicates NF-kB suppression as one
mechanism by which W. somnifera could decrease inflammation, enhance cytotoxicity and
apoptosis of tumour cells, and decrease metastasis. W. somnifera also exerts a beneficial effect
on the immune system, which may explain some of its antitumour activity10,50. It has been
reported36 that the NK cell activity is significantly enhanced by W. somnifera during
tumorigenesis. The strong immune-stimulating effect of W. somnifera elicits from macrophages
and NK cells can increase tumour cell surveillance and control.
W. somnifera (mainly its root) has been reported to possess several medicinal properties
such as antioxidant, immunomodulatary, anticonvulsant, neuroprotective, radiosenstising,
hypoglycaemic, hypolipidaemic, antiinflammatory, antitumour/anticancer etc. However, more
experimental and clinical researches should be conducted to support its therapeutic use. In
conclusion, W. somnifera reduces the cancer cell proliferation while increasing overall survival
time. Given its broad spectrum of cytotoxic and anticancer activity, W. somnifera presents itself
as a novel therapy for cancer.
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