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 Annapurna A et al. / Journal of Pharmacy Research 2011,4(4),1274-1276
Research Article
ISSN: 0974-6943 Available online through
http://jprsolutions.info
Anti-cancer activity of Curcuma longa linn.(Turmeric)
1 2 1 1
Annapurna
*1
A *, Suhasin G , Raju B Akondi , Jaya Prakash G , Siva Reddy Ch
College of Pharmaceutical Sciences, Andhra University, Visakhapatnam, A.P, India-530003
2
Department of Pharmacology and Toxicology,St.Peter’s Institute of Pharmaceutical Sciences [SPIPS],Vidya Nagar,Hanamkonda,Warangal,A.P,INDIA-506001
Received on: 05-12-2010; Revised on: 14-01-2011; Accepted on:09-03-2011

ABSTRACT
Curcuma longa Linn. (Turmeric) is used extensively in Indian cuisine and now proved to be useful in treating various types of cancers, diabetic wounds, biliary disorders etc. So the anticancer
activity of turmeric was evaluated prophylactically and therapeutically (as pre-induction treatment and post-induction treatment) against the MNU induced mammary tumors. The anticancer
activity was assessed using latency period, tumor incidence, tumor burden, tumor volume, tumor growth inhibition, histology and hematological parameters. Oral administration of turmeric
showed anticancer activity in a dose dependent manner and it was more in pre-induction treatment than in-post induction treatment groups. Topical application of turmeric was found to be more
effective in pre-induction treatment and topical treatment was more effective when compared to oral treatment. Chemo-preventive role of turmeric was more compared effective than therapeutic
role of turmeric.

Key words: Curcuma longa,anticancer activity,

INTRODUCTION
The dried Curcuma longa Linn., is the source of the spice turmeric, has been used extensively
in India for its medicinal and non medicinal uses like coloring agent, flavoring agent. Its anti
inflammatory property is known to ancient Indians and Chinese. It is also used for the
treatment of sprains and swelling caused by injury (1), for the treatment of biliary disorders,
anorexia, cough, diabetic wounds, hepatic disorders, rheumatism and sinusitis (2). It also
shows adjuvant chemoprotection in experimental stomach and oral cancer models of Swiss
mice and Syrian golden hamsters (3). Curcumin shows anti-tumour (4,5,6) and anti-
carcinogenic (7, 8, 9, 10).
Turmeric was already proved beneficial in various types of cancers like duodenal tumors (4),
tongue carcinoma (11), colon cancer (12), human breast cancer cells in-vitro (13, 14, 15, 16),
mammary tumor in-vivo (17, 18).
Few studies reported the chemopreventive effect of Curcumin. Curcumin topical application
inhibited DMBA-induced /TPA promoted skin tumors (19). So, it is logical to study the
chemo-preventive and therapeutic effects of turmeric by different routes of administration. Since
many dietary and non-dietary phytochemicals do not affect survival of normal cells and also
possess anti-tumourigenic activities, it could be a rational approach to examine the effect of
turmeric on MNU-induced mammary tumors in rats. Therefore, the present study was con-
ducted to evaluate the effect of turmeric on MNU-induced mammary tumors in rats.
MATERIALS AND METHODS:
Plant Material
Fresh raw turmeric (Curcuma longa) rhizomes were obtained from fields of Paderu,
Visakhapatnam district of Andhra Pradesh, during September 2005.They were cut down in to
small pieces, shade dried and powdered mechanically. Turmeric powder was suspended in 1%
sodium carboxy methyl cellulose and administered to rats at two dose levels orally (500mg /
Kg/B.W and 1000mg / Kg/ B.W) and topically (200mg / rat ) twice daily for 24 weeks after
MNU treatment.
Chemicals
N- methyl -N- nitrosourea (MNU) was obtained from sigma Chemical Co, St. Louis, MO,
USA. Upon arrival MNU was stored at – 200 c in the dark. All other chemicals used were of
analytical grade.
Animals
Female Sprague-Dawley rats of 4-weeks old were obtained from Mahavir Enterprises, Hyderabad.
The animals were housed in plastic cages, maintained temperature (22+/-20C) and humidity
(60+10%). Rats were maintained under 12h light/12h dark cycle and fed commercial diet
obtained from Rayan’s Bio- technology PVT – Ltd., ad libitum and water freely throughout
the study.
Induction of Mammary Carcinogenesis
For systemic exposure, MNU was dissolved immediately prior to its use in 0.9% Nacl
(Normal saline) containing 0.05% acetic acid. The intra peritoneal injection of 50mg / kg
body weight, a dose known to produce high incidence of mammary cancer (20) was made
alongthe ventral midline of the 50 days old rats.
*Corresponding author.
Annapurna Akula,
University College of Pharmaceutical Sciences,
Andhra University, Visakhapatnam,
A.P, India-530003
Journal of Pharmacy Research Vol.4.Issue 4. April 2011
Experimental Design:
The SD rats were randomly divided in to 7 groups. Number of animals varies from group to
group and it ranges from 12 animals to 24 animals. The details were given as follows. Group
1 (n = 15), Control group received MNU only, Group 2 (n = 12), Prophylactic treatment of
turmeric suspension (500mg/kg, P.O), Group 3 (n = 24), Prophylactic treatment of turmeric
suspension (1000mg/kg, P.O), Group 4 (n = 24), Prophylactic treatment of turmeric suspen-
sion (200mg/rat, topical), Group 5 (n = 12), Therapeutic treatment of turmeric suspension
(500mg/kg, P.O), Group 6 (n = 12), Therapeutic treatment of turmeric suspension (1000mg/
kg, P.O), Group 7 (n = 12), Therapeutic treatment of turmeric suspension (200mg/rat, topical).
Raw turmeric powder was administered as suspension (Suspended in 1%Sodium carboxy
methyl cellulose) by two routes, orally and topically in different groups. Turmeric was
administered orally at two doses 500 mg/kg and 1000mg/kg. In topical treatment group, the
animals were shaved ventrally by using hair removing cream and turmeric suspension was
applied along the ventral side of rat by using a small brush in dose of 200 mg / rat, twice daily.
Pre-induction treatment:
Animals of groups 2 and 3 were treated with oral administration of 500 mg/kg and 1000 mg/
kg body weight of turmeric (group1: 500mg/kg body weight; group 2: 1000mg/kg body
weight) and animals of group 4 were treated with 200mg/rat turmeric topical application twice
daily from two weeks before the carcinogen treatment and continued till the end of the study (for
24 weeks after MNU treatment).
Post-induction treatment:
Animals of group 5 and 6, after 2 weeks of MNU administration were treated with oral
administration of 500 mg/kg bodyweight and 1000 mg/kg bodyweight of turmeric (group 5:
500 mg/kg body weight; group 6:1000mg /kg body weight ) and animals of group 7 were
treated with 200mg/rat turmeric topically twice daily from two weeks before the carcinogen
treatment and continued till the end of the study (for 24 weeks after MNU treatment) Follow-
ing carcinogen treatment, all rats were weighed once weekly and palpated for detection of
mammary tumors. The rats were sacrificed 24 weeks after MNU treatment. Liver and uterus
were dissected out from each of the experimental animal and weighed. The Latency period, the
number of tumor bearing animals (% incidence), the number of tumors per animal (multiplic-
ity) and tumor burden (weight/animal) were determined.
The tumor volume (V=4/3 πr3),
( Vc − Vt )
tumor growth inhibition (TG1 %= × 100) was calculated.
Vc
Hemoglobin content, RBC, WBC and platelet counts were recorded every month starting from
the experiment till the end of the experiment.
Statistical analysis
The difference in tumor volumes, latency periods, body weights, wet uterus and wet liver
weights were statistically evaluated by One–way analysis of variances (ANOVA), followed by
Tukey’s t test and the tumor incidence rates were determined using chi–square test. Values of
P<0.001, P<0.01, P<0.05 were considered significant.
RESULTS:
There was 20% mortality in control group animals whereas mortality has been significantly
reduced in prophylactic and therapeutic treatment of groups (Table 1).
1274-1276
 Annapurna A et al. / Journal of Pharmacy Research 2011,4(4),1274-1276
Effect of turmeric on percent incidence rates of mammary tumors:
Incidence rate of mammary tumors in control group was 83.33%. Percent incidence rates after
treatment were given in Figure 1 and Table 1. However, incidence rates were significantly
decreased in both prophylactic & therapeutic treatment of turmeric suspension. Prophylactic
treatment of turmeric has superiorly reduced the incidence rates of mammary tumor when
compared to therapeutic treatment of turmeric. Hence it is evident that per oral administration
and topical application of turmeric was more effective when administered prophylactically than
therapeutically in reducing the incidence rates when compared to control and it was dose
dependent.
100
topically has significantly reduced the mean tumor volume. Interestingly, prophylactic treat-
ment of turmeric at a dose of 200mg/kg topically was most effective than prophylactic oral
treatment of turmeric and both oral and topical therapeutic treatment. Therapeutic treatment of
turmeric at a dose of 500mg/kg (P<0.01) and 1000mg/kg (P<0.001) has significantly reduced
mean tumor volume but not on par with the prophylactic treatment. In addition, prophylactic
topical application of turmeric has significantly (P<0.001) reduced the mean tumor volume
when compared to therapeutic topical application of turmeric has which no significant effect on
mean tumor volume. Results of mean tumor volume were given in Figure 3 and Table 2.
30
***

(Weeks) Mean ± SEM

Mean Latency pe riod
*** ***
Control
83.33%
20
Mean ± SEM (n = 12-24)

80 Turmeric 500 mg/kg,p.o.(Prophylactic)

***
Turmeric1000 mg/kg,p.o.(Prophylactic)
% Incidence rate

Control
63.3% Turmeric 200 mg/rat, topical (Prophylactic)
60% Turmeric 500 mg/kg,p.o.(Prophylactic)
60 10
Turmeric 1000 mg/kg,p.o.(Prophylactic) Turmeric 500 mg/kg,p.o.(Therapeutic)
Turmeric 1000 mg/kg,p.o.(Therapeutic)
Turmeric 200 mg/rat, topical (Prophylactic)
40 36.3% Turmeric 200 mg/rat, topical (Therapeutic)
Turmeric 500 mg/kg,p.o.(Therapeutic) 0
25% Turmeric 1000 mg/kg,p.o.(Therapeutic)
20 Turmeric 200 mg/rat, topical (Therapeutic) ***p<0.001 vs Control group by One way ANOVA/Tukey's test
8.33% 10% Figure 3: Effect of prophylactic and therapeutic treatment mean tumor volume in female SD rats.
0 Table 2: Effect of prophylactic and therapeutic treatment on parameters of induced mammary
cancer in female SD rats
***p<0.05 vs Number of rats without tumors by ψ Chi square test 2

Animal Groups Total No. No. of No. of Total *Tumor Mean TGI %
Figure 1: Effect of prophylactic and therapeutic treatment of turmeric on incidence rate of of rats effective rats with No. of burden tumor
mammary tumors in female SD rats. rats tumors tumors volume

Table 1: Effect of prophylactic and therapeutic treatment of turmeric on the incidence of Control 15 12 10 17 1.41 3.21 —
Turmeric 500 mg/kg, p.o. 12 12 3 4 0.33 1.10 65.73
tumor development in female SD rats.
(Prophylactic)
Turmeric 1000 mg/kg, p.o. 24 24 2 1 0.08 0.86 73.20
Animal Groups Total No. No. of % No. of No. of Incidence (Prophylactic)
of rats effective Mortality rats without rats with rates Turmeric 200 mg/kg, topical 24 20 2 2 0.20 0.23 92.83
rats tumors tumors (Prophylactic)
Turmeric 500 mg/kg, p.o. 12 10 6 10 1.00 2.02 37.07
Control 15 12 20 2 10* 83.33% (Therapeutic)
Turmeric 500 mg/kg, p.o. 12 12 0 9 3* 25% Turmeric 1000 mg/kg, p.o. 12 11 4 5 0.45 1.69 47.35
(Prophylactic) ( Therapeutic )
Turmeric 1000 mg/kg, p.o. 24 24 0 22 2* 8.33% Turmeric 200 mg/kg, topical 12 11 7 13 1.18 2.56 20.24
(Prophylactic) ( Therapeutic )
Turmeric 200 mg/kg, topical 24 20 16 18 2* 10%
(Prophylactic) *Tumor burden = Number of tumors per rat; Tumor growth inhibition (TGI%) = (Vc-Vt)/Vc
Turmeric 500 mg/kg, p.o. 12 10 16 4 6* 60% × 100 ; Vc = mean tumor volume in control group at any point of time, Vt = mean tumor volume
(Therapeutic) of treatment group at any point of time; Significance was given in the graphs for mean tumor
Turmeric 1000 mg/kg, p.o. 12 11 8.33 7 4* 36.36%
( Therapeutic ) volume and tumor growth inhibition in Figure 3 and figure 4 respectively.
Turmeric 200 mg/kg, topical 12 11 8.33 4 7* 63.36%
( Therapeutic ) Effect of turmeric on percent tumor growth inhibition (% TGI):
Percent tumor growth inhibitions of prophylactic turmeric 500mg/kg, 1000mg/kg P.O. and
P< 0.05, * statistically significant when compared to number of rats without tumors. Ψ Chi 2
prophylactic turmeric 200mg/kg topical application were found to be 65.73, 73.20 and 92.83
square test; S = Statistically significant; NS = Statistically insignificant respectively. Hence, prophylactic treatment of turmeric orally has shown percent tumor growth
Effect of turmeric on mean latency periods for induction of mammary tumors: inhibition dose dependently. The major finding of the study is that the highest degree of
Prophylactic treatment of turmeric when given orally showed dose dependent and significant percent growth inhibition with prophylactic topical application of turmeric (92.83) which is
beneficial effect. Curcumin prolonged the mean latency periods for induction of mammary significantly (P<0.001) higher than the TGI of prophylactic turmeric treatment with 500mg/
tumors. Similarly, Prophylactic treatment of turmeric when used topically at a dose of 200mg/ kg.p.o. Percent tumor growth inhibitions of therapeutic turmeric 500mg/kg, 1000mg/kg P.O.
kg has significantly increased the mean latency periods. Interestingly, prophylactic treatment and therapeutic turmeric 200mg/kg topical application were found to be 37.07, 47.35 and
of turmeric at a dose of 200mg/kg topically was most effective than prophylactic oral treatment 20.24 respectively. In contrast to prophylactic topical application of turmeric, therapeutic
and oral and topical therapeutic treatment. Therapeutic treatment of turmeric has no significant topical application of turmeric has shown lesser degree of percent tumor growth inhibition.
effect on mean latency periods except at 1000 mg/kg, P.O. (P<0.001). In addition, prophylac- Results were given in the Figure 4 and Table 2.
tic topical application of turmeric has significantly (P<0.001) prolonged the mean latency
periods when compared to therapeutic topical application of turmeric which has no significant
effect on mean latency periods. Results of mean latency periods were given in Figure 2. 4
Mean Tumor volume (Cm 3 )

Control
30
Turmeric 500 mg/kg,p.o.(Prophylactic)
*** 3
(Weeks) Mean ± SEM

Turmeric 1000 mg/kg,p.o.(Prophylactic)

Mean Latency pe riod

Mean ± SEM

*** ***
Control ** Turmeric 200 mg/rat, topical (Prophylactic)
20 Turmeric 500 mg/kg,p.o.(Prophylactic) 2 *** Turmeric 500 mg/kg,p.o.(Therapeutic)
***
Turmeric1000 mg/kg,p.o.(Prophylactic) *** *** Turmeric 1000 mg/kg,p.o.(The rapeutic)
Turmeric 200 mg/rat, topical (Prophylactic) 1 Turmeric 200 mg/rat, topical (Therapeutic)
10 ***
Turmeric 500 mg/kg,p.o.(Therapeutic)
Turmeric 1000 mg/kg,p.o.(Therapeutic) 0
Turmeric 200 mg/rat, topical (Therapeutic)
**p<0.01, ***p<0.001 vs Control group by One way ANOVA/Tukey's te st
0
Figure 4: Effect of prophylactic and therapeutic treatment on percent tumor growth inhibition
***p<0.001 vs Control group by One way ANOVA/Tukey's test in female SD rats.
Figure 2: Mean latency periods across all (control and prophylactic, therapeutic treatment) the
groups of female SD rats
The difference between the body weights, wet uterus, and wet liver weights between control
Effect of turmeric on mean tumor volume: and treated groups were statistically insignificant (data was not shown). It was observed that
Prophylactic treatment of turmeric when given orally, reduced the mean tumor volume there was no statistically significant difference in hematological parameters including WBC,
significantly (P<0.001). Similarly, Prophylactic treatment of turmeric at a dose of 200mg/kg RBC and hemoglobin values between control and treated groups. Hence it was evident that

Journal of Pharmacy Research Vol.4.Issue 4. April 2011 1274-1276

 Annapurna A et al. / Journal of Pharmacy Research 2011,4(4),1274-1276
there was no bone marrow depression with turmeric treatment. Results of hematological Till date, there was no evidence of anticancer activity with topical application of turmeric in
parameters, WBC, RBC and RBC were given in Table 3, Table 4 and Table 5. breast cancer model. In two in vivo studies reported earlier, topical application of 100 or 3000
Table 3: Effect of prophylactic and therapeutic treatment for six months on WBC (103/µL) nmol curcumin in CD-1 mice and 0.2% or 1% curcumin in diet significantly reduced the tumor
incidence and tumor volume in dimethyl benz (a) anthracene (DMBA) initiated and 12,0-
count in female SD rats
tetradecanoylphorbal -13-acetate (TPA) promoted skin tumors (22).
Animal Groups 0 time 1st Month 2ndMonth 3rd Month 4th Month 5th Month 6th Month

Control 6.9 ±0.36 8.1±0.62 8.34±0.36 8.26±0.46 8.43±0.38 8.60±0.42 8.82±0.21 The general anti-carcinognic effect of Curcumin involves the mechanisms like induction of
Turmeric 500 mg/kg, p.o. 7.02±0.46 8.6±0.69 8.46±0.52 8.47±0.38 8.56±0.26 8.72±0.46 8.97±0.42 apoptosis and inhibits cell-cycle progression, both of which are instrumental in preventing
(Prophylactic) cancerous cell growth in rat aortic smooth muscle cells (23). The antiproliferative effect is
Turmeric 1000 mg/kg, p.o 7.60±0.51 8.30±0.64 8.48±0.47 8.56±0.82 8.68±0.43 8.86±0.48 8.96±0.33
.(Prophylactic) mediated partly through inhibition of protein tyrosine kinase and c-myc mRNA expression and
Turmeric 200 mg/kg, topical 6.49±0.45 7.54±0.59 7.92±0.54 8.28±0.57 8.65±0.52 8.71±0.37 8.79±0.27 the apoptotic effect may partly be mediated through inhibition of protein tyrosine kinase,
(Prophylactic) protein kinase C, c-myc mRNA expression and bcl-2 mRNA expression (23). Specifically,
Turmeric 500 mg/kg, p.o 6.36±0.26 7.89±0.43 7.99±0.38 8.03±0.48 8.16±0.61 8.11±0.34 8.3±0.36
.(Therapeutic) Curcumin suppresses human breast carcinoma through multiple pathways. Its antiproliferative
Turmeric 1000 mg/kg, p.o. 7.12±0.32 7.90±0.60 8.13±0.42 8.15±0.39 8.34±0.44 8.48±0.41 8.65±0.21 effect is estrogendependent in ER (estrogen receptor)-positive MCF-7 cells and estrogen-
( Therapeutic ) independent in ER-negative MDA-MB-231 cells (9). Curcumin also down regulates matrix
Turmeric 200 mg/kg, topical 7.32±0.52 7.92±0.52 8.22±0.48 8.34±0.40 8.58±0.39 8.73±0.31 8.68±0.46
( Therapeutic ) metalloproteinase (MMP)-2 and upregulates tissue inhibitor of metalloproteinase (TIMP)-1,
two common effector molecules involved in cell invasion (9). It also induces apoptosis
Table 4: Effect of prophylactic and therapeutic treatment for six months on RBC (106/µL) through P53-dependent Bax induction in human breast cancer cells (10).
count in female SD rats
Since major side effect of anti-cancer drugs is bone marrow depression, the present study has
Animal Groups 0 time 1st Month 2nd Month 3rd Month 4th Month 5th Month 6th Month
investigated the effect of chronic turmeric treatment on hematological parameters. There was no
Control 4.80±0.15 6.71±0.32 6.93±0.61 7.41±0.53 7.60±0.24 7.67±0.36 7.80±0.32 significant difference in hematological parameters among the different treatment groups and
Turmeric 500 mg/kg, p.o. 4.67±0.18 6.24±0.25 6.46±0.53 6.98±0.48 7.26±0.62 7.41±0.41 7.64±0.41 control group. Hence it was evident that no bone marrow depression with turmeric treatment
(Prophylactic) was observed, which is a major side effect with cytotoxic chemotherapy.
Turmeric 1000 mg/kg, p.o. 4.92±0.13 6.89±0.30 7.03±0.56 6.97±0.36 7.13±0.43 7.24±0.26 7.92±0.56
(Prophylactic)
Turmeric 200 mg/kg, topical 4.89±0.10 6.82±0.28 7.14±0.51 7.11±0.53 7.62±0.31 7.41±0.43 7.47±0.40 In conclusion, the turmeric acts effectively both orally and topically initiation stage of mam-
(Prophylactic) mary cancer than in the promotion stage of mammary carcinoma. This stage specificity of
Turmeric 500 mg/kg, p.o. 4.32±0.30 6.43±0.31 6.89±0.43 7.26±0.46 7.70±0.53 7.78±0.51 7.94±0.38
(Therapeutic) turmeric’s anticancer activity must be established by further investigations.
Turmeric 1000 mg/kg, p.o. 4.50±0.21 6.65±0.22 6.96±0.49 7.44±0.48 7.86±0.42 7.80±0.38 7.82±0.36
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Journal of Pharmacy Research Vol.4.Issue 4. April 2011 1274-1276

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