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com

International Journal of ISSN: 0976-9390

Chemical and Pharmaceutical Sciences
2010, Dec., Vol.1 (2)

Evaluation of the Anti-Tumor and Antioxidant Activity of

Amorphophallus Paeonifolius on DMBA Induced Mammary
Carcinoma
1
Jagatheesh K*, 2Arumugam V, 3Elangovan N and 4PavanKumar P
1
Dept of Pharmacology, CARISM SASTRA University, Tanjoore, Tamil Nadu, India.
2
Dept of Pharmacology, KMCH college of Pharmacy. Coimbatore, Tamil Nadu India.
3
Dept of Biotechnology, Periyar University. Salem, Tamil Nadu, India.
4
Dept of Pharmacology, Chebrolu Hanumaiah Institute of Pharmaceutical Sciences.Guntur,
Andrapradesh, India.
*
Corresponding author: E-Mail – kjagatheesh@gmail.com

ABSTRACT
The present investigation was carried out to investigate the anti-tumor activity,
Antioxidant activity of the ethanolic extract of Amorphophallus Paeonifolius tubers against 7,12-
dimethyl benz (a) anthracene (DMBA) induced mammary tumour in rats.The Pharmacognostic
and Phytochemical studies over Amorphophallus Paeonifolius were studied first and the anti-
oxidant activity of the same were studied In-vitro and In-vivo methods finally the plant extract
was evaluated for mammary tumour activity by chemical induced tumour.The results shows total
Flavanoids compound in Ethanolic tuber extract of Amorphophallus Paeonifolius was found to
be 8.8 g/100g calculated as Quercetin equivalent, effect of Ethanolic extract on RBC, WBC, Hb
& Neutrophils as a-P< 0.001,b-P<0.01,c-P<0.05, ns-non significant, Effect of AP-extract on
tumor latency and tumour burden were found as extremely significant at P<0.001. One-way
ANOVA Tukey method was followed for statistics. It was concluded that ethanolic extract of
Amorphophallus Paeonifolius has shown significant antitumor and antioxidant effect in animals.
Keywords: Amorphophallus Paeonifolius, Anti-Oxidant, Mammary tumour
1. INTRODUCTION cancerous). Cancer affects people at all ages
Cancer is a class of diseases in which with the risk for most types increasing with
a group of cells display uncontrolled growth age. Cancer caused about 15% of all human
(division beyond the normal limits), deaths in 2009. Breast cancer can begin in
invasion (intrusion on and destruction of different areas of the breast – the ducts, the
adjacent tissues), and sometimes metastasis lobules, or in some cases, the tissue in between.
(spread to other locations in the body via There are many different types of breast cancer,
lymph or blood). These three malignant with different stages (spread), aggressiveness,
properties of cancers differentiate them from and genetic makeup; survival varies greatly
benign tumors, which are self-limited, and depending on those factors. [3]
do not invade or metastasis. [1, 2] Tumors can It is significant that over 60% of
be malignant (cancerous) or benign (non currently used anti-cancer agents are derived in
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one way or another from natural sources,
including plants, marine organisms and
micro-organisms. Indeed, these sources have
played, and continue to play, a dominant
role in the discovery of leads for the
treatment of most human diseases. [3]
The search for anticancer agents
from plant sources started in 1950s with the
discovery and development of vinca
alkaloids, vincristine, vinblastine and the
isolation of cytotoxic podophyllotoxin.
These discoveries in turn lead to discovery
of taxanes and camptothecins but their
development into clinically active agent
spanned period of 30 years from 1960s to
1990s.[4,5]
The plant Amorphophallus
paeonifolius (AP) synonym
Amorphophallus campanulatus belongs to
the family of Araceae Elephant foot yam is
basically a crop of Southeast Asian origin. It
grows in wild form in the Philippines,
Malaysia, Indonesia, and other Southeast
Asian countries. In India it is grown mostly
in Kerala, Andhra Pradesh, Maharashtra and
Orissa. A stout herbaceous plant""with
underground hemispherical depressed dark
brown corm; leaves compound, large,
solitary, petiole stout, mottled, 60-90 cm
long, leaflets 5-12.5 cm long of variable
width and the corms contain betulinic acid,
ß-sitosterol, stigmasterol, triacontane and ß-
sitosterol palmitate. Besides these, glucose,
galactose, rhamnose and exylose are also
present. The corms are irritant due to the
composition of arginine, histidine, leucine,
isoleucine, lysine, methionine,
phenylalanine, threonine, tryptophan, and
valine.
They are useful in vitiated conditions
of vata and kapha, rthralgia, ephantiasis,
tumours, inflammations, haemorrhoids,
haemorrhages, vomiting, cough, onchitis,
asthma, anorexia, dyspepsia, flatulence,
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colic constipation, helminthiasis hepatopathy,
splenopathy, amenorrhoea, dysmenorrhoea,
seminal weakness, fatigue, anemia and general
debility. [5- 8]
2. MATERIALS AND METHODS
Plant materials [tubers] were locally
collected and authenticated by Botanical
Survey of India (BSI), Southern circle,
Coimbatore, Tamilnadu
2.1 Extraction of tuber of Amorphophallus
paeonifolius
The tuber of Amorphophallus
Paeonifolius was cut into pieces of around 5
mm, shade dried and ground to powder. 300
gm of powdered tuber was extracted with
petroleum ether to remove fatty material for 10
cycles. The material was air dried and then
extracted with chloroform as said above, the
tuber material was subjected to dry and finally
ethanolic extraction was done. The extract were
concentrated to dryness and stored in vaccum
dessicator.
2.2 Phytochemical analysis of ethanolic
extracts of Amorphophallus Paeonifolius
The photochemical analysis of the plant
with standard chemical tests was shown that
extract contains alkaloids, glycosides,
carbohydrates , steroids and sterols, Tannins,
proteins, peptides or amino acids and aromatic
amino acid
2.2.1. Determination of Total Flavanoids
Flavones and flavanols in the ethanolic
extracts of Amorphophallus Paeonifolius were
estimated as Quercetin equivalent. Quercetin
was used to make the calibration curve [10, 20,
30, 40, 50, 60, 70, 80, 90 and 100μg/ml in
99.9% ethanol (v/v)]. The standard solutions or
extracts (0.5 ml) were mixed with 1.5 ml of
95% ethanol (v/v), 0.1 ml of 10% aluminium
chloride, 0.1 ml of 1 mol/l sodium acetate and
2.8 ml water. The volume of 10% aluminum
chloride was substituted by the same volume of
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distilled water in blank. After incubation at
room temperature for 30 min, the
absorbance of the reaction mixture was
measured at 415 nm. The mean of three
readings was used and the total Flavanoids
content was expressed in milligrams of
Quercetin equivalents 190.2 mg/g extract.
The coefficient of determination was
r2 = 0.9985. [9]
2.3. In vitro anti-oxidant activity
The in vitro anti-oxidant activity of
the extract was done by various available
methods as follows
2.3.1. DPPH (Diphenylpicrylhydrazyl)
Radical scavenging activity
The scavenging activity of
Amorphophallus Paeonifolius extract was
measured in terms of hydrogen donating or
radical scavenging ability using the stable
radical DPPH Blois et al., (1958).[10] 0.1Mm
solution of DPPH in ethanol was prepared
and 1.0ml of this solution was added to 3 ml
of extract solution and standard in water at
different concentrations (10-100µg/ml). 30
min later absorbance was measured at
517nm. Lower absorbance of the reaction
mixture indicated higher free radical
scavenging activity. The capability to
scavenge the DPPH radical was calculated
using the following equation
2.3.2. Reducing power
The reductive potential of the extract
was determined according to the method of
Oyaizu et al., (1986).[11] The different
concentration of extracts and standard in
1ml of distilled water was mixed with
phosphate buffer (2.5ml, 0.2m, pH6.6) and
potassium ferricyanide [K3 Fe(CN)6]
(2.5ml,1% w/v). The mixture was incubated
at 50oC for 20min.2.5ml of
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trichloroaceticacid (10%w/v) was added to the
mixture, which was then centrifuged for 10 min
at 1000rpm. The 2.5 ml of supernatant was
mixed with distilled water (2.5ml) and Fecl3
(0.5ml, 0.1% w/v), and mixed. Absorbance was
measured at 700nm in a spectrophotometer.
Higher absorbance of the reaction mixture
indicated greater reductive potential.
2.3.3. Superoxide anion scavenging activity
assay
The scavenging activity of
Amorphophallus Paeonifolius towards
superoxide anion radicals was measured by the
method of Ni-shimiki et al.,(1972). [12] and
slightly modified. About 1ml of nitroblue
tetrazolium solution (156µm in 100Mm
phosphate buffer, pH 7.4) and 0.1ml of
different concentrations of extract and standard
in water were mixed. The reaction was initiated
by adding 100µl of
phenazinemethosulphate(PMS) solution
(60µM) in 100Mm phosphate buffer(pH7.4) to
the mixture. The reaction mixture was
incubated at room temperature for 5 min and
the absorbance at 560nm was measured against
reagent blank in spectrophotometer. (Quercetin
was used as standard).
2.3.4. ABTS radical scavenging activity
assay
The ABTS radical scavenging activity
of the extract was measured by Rice-Evans
et.al. (1997).[13]ABTS radical cation (ABTS+)
was produced by reacting ABTS solution
(7mm) with 2.45 Mm ammonium persulphate
and the mixture were allowed to stand in dark
at room temperature for 12-16hrs before use.
Different concentration (20-100µg/ml) of
ethanolic extract and standard (0.5ml) were
added to 0.3ml of ABTS solution and the
ethanol to make 1ml. Quercetin was used as
standard. The absorbance was read at 745nm.
The experiment was performed in triplicate.
2.4. Pharmacological Evaluation
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2.4.1. Acute toxicity Studies
Healthy adult Wistar albino mice of
either sex, starved overnight were divided
into five groups (n=6) and were orally fed
with the Ethanolic extract of
Amorphophallus Paeonifolius in increasing
dose levels of 100, 500, 1000, 2000mg/kg
body weight respectively and they were
observed for behavioral, neurological and
autonomic profiles and after a period of 24
and 72 hr they were observed for any
lethality.
2.4.2. Induction of cancer
The cancer was inducted by using
chemical induction method using 7, 12-
Dimethyl Benz (A) Anthracene(DMBA)
DMBA was purchased from sigma
chemicals, Mumbai, India. 120 mg DMBA
was dissolved in 24 ml olive oil. It forms a
yellow colour solution. DMBA solution was
given to animals orally with the help of oral
feeding needle at a dose of 25 mg/kg.
2.5. In-vivo anti-oxidant activity
All the experimental animals were
killed by cervical decapitation after the
experimental period. For the estimation of
non-enzymic and enzymic antioxidants,
tissue (liver and kidney) was minced and
homogenized (10% w/v) in 0.1 M phosphate
buffer (Ph 7.0) and centrifuged for 10 min
and the resulting supernatant was used for
enzyme assays.
2.5.1. Enzymatic Anti-oxidant activity
2.5.1.1. Estimation of catalase activity
Catalase activity was measured by
method described by Sinha et.al.(1972).[14]
The reaction mixture (1.5ml, vol) contained
1.0ml of 0.01M phosphate buffer (pH7.0),
0.1ml of tissue homogenate and 0.4ml of
2M H2O2.The reaction was stopped by the
addition of 2.0ml of dichromate-acetic acid
reagent (5% pot. dichromate and glacial
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acetic acid were mixed in 1:3 ratio). Then the
absorbance was read at 620nm; CAT activity
was expressed as µMol of H2O2
consumed/min/mg protein.
2.5.1.2. Estimation of Superoxide dismutase
(SOD) Activity
The activity of superoxide dismutase
was assayed by the method of Kakkar
et.al..(1984).[15] Breifly in a test tube; 0.5ml of
supernatant tissue homogenate was taken. To
this 1.5ml of carbonate buffer(pH10.2),0.5ml of
0.1Mm EDTA and 0.4ml of epinephrine was
added and the OD was taken at
480nm.Epinephrine was added just before
taking the OD. This activity is to be expressed
as units/min/mg protein.
2.5.1.3. Estimation of Glutathione
peroxidase (GPx) Activity
GPx activity was measured by method
described by Rotruck et.al.(1973).[16] Briefly,
the reaction mixture contained 0.2ml 0.4M
Phosphate buffer (pH7), 0.1ml 10Mm sodium
azide, 0.2ml tissue homogenized in 0.4M
Phosphate buffer(pH7), 0.2ml glutathione, and
0.1ml 0.2Mm H2O2. The contents were
incubated for 10min at 37o C, 0.4ml 10% TCA
was added to stop the reaction and centrifuged
at 3200 rpm for 20min.The supernatant was
assayed for glutathione content using Ellman’s
reagent(19.5g 5,5’-dithiobisnitrobenzoic acid
(DTNB) in 100ml 0.1% sodium citrate). The
activity is to be expressed as µMol of GSH
consumed/min/mg protein.
2.5.2. Non-enzymatic anti-oxidant activity
2.5.2.1. Estimation of reduced glutathione
(GSH) activity
GSH was determined by the method of
Ellman et al., (1959). [17] A known weight of
tissue homogenized in phosphate buffer from
this 0.5ml was pipetted out and precipitated
with 2ml of 5% TCA 1ml of the supernatant
was taken after centrifugation at 3200 rpm for
20min and added to it 0.5ml of Ellman’s
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reagent and 3ml of phosphate buffer (pH 2.6.4. Estimation of protein

8.0). Then the absorbance was read at Procedure described by Lowery et.al.
412nm. The values were expressed as (1951). [18] was used for protein estimation. The
mg/100 g tissue. method was based on the biuret reaction,
2.6. Anti –tumor activity formation of a protein-copper complex and
2.6.1. Experimental design for treatment reduction of phosphomolybdo tungstate reagent
oriented study (Folin-ciocalteu phenol reagent) by tyrosine
and tryptophan residues of protein to form a
Experimental rats were divided into coloured product.
5 groups of six animals each and received
the following treatment for 90 days. Group 2.7. Statistical Analysis
I- Control rats given only Saline (p.o.), The data for various parameters were
Group II- Rats given DMBA only 25 mg/kg analyzed using analysis of variance (ANOVA)
(p.o.), Group III- Rats treated with DMBA + Tukey, followed by compared of all pairs of
AP tuber extract 50 mg/kg (p.o.), Group IV- column.
Rats treated with DMBA + AP tuber extract 3. RESULTS AND DISCUSSION
100mg/kg (p.o.), Group V- Rats treated with
DMBA + TAM 10 mg/kg (p.o.) Table 1: Determination of Total Flavanoids.
Concentration
2.6.2. Hematological studies Sample Absorbance
(μg/ml)
In this 90 days study both group of 10 0.2651
animals were treated with respective extracts 20 0.3663
and standard drugs via oral route. Body Quercetin as 30 0.5014
weight was taken every week till 17th week. standard
40 0.8116
After the completion of treatment, blood
50 0.9934
was collected for hematological parameters
Ethanolic tuber
estimation like RBC, WBC (Total and extract of
250 0.4942
differential count by Improved Neubauer’s Amorphophallus
Paeonifolius
counting chamber method using Haeyem’s
fluid) Hb (by sahli’s Haemoglobinometer
method) and Neutrophil by Improved Total Flavanoids compound in
Neubauer’s counting chamber method. Ethanolic tuber extract of Amorphophallus
2.6.3. Bio chemical parameters Paeonifolius was found to be 8.8 g/100g
calculated as Quercetin equivalent
Aliquot of blood was collected and
centrifuged at 5000 rpm for 5 min to Preliminary phytochemical screening
separate the serum. This serum was used for indicated the presence of alkaloids and
estimation of SGOT by Optimized UV- test flavanoids in Amorphophallus Paeonifolius
according to IFCC(International Federation tuber extract. flavanoids (8.8 g/100 g)
Of Clinical Chemistry and Laboratory calculated as Quercetin equivalent. Flavanoids
Medicine), SGPT by Kinetic UV test, have been shown to possess anti-mutagenic and
according to the International Federation of anti-malignant effects. Moreover, flavanoids
Clinical Chemistry and Laboratory have an antitumor role through their effects on
Medicine, serum creatinine by Jaffe Method signal transduction in cell proliferation and
(modified), serum urea by Urease-GLDH angiogenesis. The cytotoxic and antitumor
enzymatic UV test method .
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properties of the extract may be due to these
compounds.
DPPH is a stable free radical at room
temperature and accepts an electron or
hydrogen radical to become a stable dia-
magnetic molecule .The reduction capability
of DPPH was determined by the decrease in its
absorbance at 517 nm, which is induced by
anti-oxidants. On the DPPH radical, AP extract
significant scavenging effects when compared
with that Quercetin.(table 1)

Table 2: In Vitro Anti-Oxidant Activity

Amorphophallus Paeonifolius Tuber extract Quercetin Standard
Concentration(μg/ml) % Inhibition Concentration(μg/ml) % Inhibition
10 35.40 10 42.75
20 45.15 20 55.11
DPPH radical 30 53.06 30 65.18
scavenging activity 40 53.42 40 66.33
50 53.48 10 70.56
60 54.78 20 72.19
20 65.35 20 81.03
40 67.47 40 81.85
ABTS radical
scavenging activity 60 69.81 60 82.04
assay
80 71.79 80 82.56
100 73.58 100 86.96
40 61.93 40 87.76
Superoxide anion 60 62.24 60 87.99
scavenging activity
assay 80 62.40 80 88.66
100 63.07 100 88.76

In ABTS scavenging activity the
extract tend to possess significant
scavenging activity in comparison to
Quarcetin. The decolorization of ABTS+
radical is an unambiguous way to measure
the antioxidant activity of phenolic
compounds. The determination of phenolic
antioxidant using the oxygen radical
absorbance capacity (ORAC), ABTS+ and
the 1, 1-diphenyl 2-picrylhydrazyl (DPPH)
assays. Thus monitoring the antioxidant
activity by ABTS+ radical scavenging assay
gives good prediction of their ORAC and
DPPH radical scavenging capacity. AP
Extract. showed potential activity in ABTS+
decolorization. 73% inhibition was noted
with 100μg/ml of AP extract. Decolorization
of ABTS+ in the present study reflects the
capacity of an antioxidant species to donate
electrons or hydrogen atoms to inactivate this
radical cation.(table 2)
Indirect stimulation of lipid oxidation
by superoxide as a result of superoxide acts as
precursors of singlet oxygen radical. This free
radical responsible for cellular damages and
oxidative degradation of macromolecules. The
scavenging effect of AP extract and Quercetin
on superoxide radical was compared. On the
superoxide radical, AP extract has significant
scavenging effects when compared with that
Quercetin.
The acute toxicity studies revealed the
non-toxic nature of the ethanolic extract of AP
up to higher concentration of 2000mg/kg, there
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was no lethality or any toxic reaction found
with the selected dose until the end of the
study period. The dose of the test drug has
been selected on the basis of dose
calibration curve.
The result of the study have shown that
the ethanolic extract of AP at a dose of 50
mg/kg and 100 mg /kg has a marked significant
on DMBA mammary carcinoma rats.

Table 3: In-Vivo Anti-Oxidant Activity

Enzymatic antioxidant activity
Group (n=6) control DMBA only DMBA + DMBA + DMBA +
AP-Tuber AP-Tuber Tamoxifen
extract(50mg/kg) extract(100mg/kg) (10mg/kg)

Estimation of
Liver 317.4±70 287.9±10a 443.7±27.8 a 602.5±21.21 a 483.3±29.63 a
Catalase Activity
Kidney 279.13±37.55 189.4±38.11a 540.78±30.8 a 717.69±33.9 a 827.02±22.88 a
Estimation of Liver 40.84±8.56 4.22±2.31a 31.63±3.35 a 53.42±4.02 a 63.82±7.17 a
Superoxide a b ns
Kidney 36.6±5.92 16.35±1.85 30.29±3.36 24.22±9.20 73.41±1.28 a
Dismutase (SOD)
Activity
Estimation of Liver 206.04±28.88 97.41±11.7a 132.64±43.89 a 290.66±17.09 a 230.94±8.27 a
Glutathione
Kidney 269.71±11.47 87.37±13.12a 227.23±39.98 a 260.45±36.80 a 291.59±6.00 a
peroxidase (GPx)
activity
Non-enzymatic antioxidant activity

Estimation of Liver 851.47±105.2 358.36±59.47a 907.66±200.9c 1034.03±44.52 a 1015.20±166.8 a

reduced a a a
Kidney 837.95±94.7 400.74±68.84 1041.61±55.69 1136.86±93.07 1161.12±112.4 a
glutathione (GSH)

a-P< 0.001,b-P<0.01,c-P<0.05, ns-non significant other to hydrogen peroxide. Hydrogen

Data was expressed as mean ±SD.(n=6
animals in each group). Values were statistically
extremely significant at P<0.001. Unit: unit/mg of
protein
One way ANOVA Tukey, followed
by comparison of all pairs of columns test
was performed. DMBA only (group 2)
compared with control (group 1), AP-Tuber
extract 50 mg/kg (Group3), AP-Tuber
extract 100 mg/kg (group 4), TAM 10
mg/kg (group 5) were compared
The antioxidant system contributes
towards the inhibition of carcinogenesis.
SOD, a pair of Superoxide anions by
oxidizing one to oxygen and reducing the
peroxide, which is also considered to be
mutagenic, degraded to H20 and oxygen by
Catalase48. The decrease in the level of SOD
and Catalase in only DMBA treated animals
and an increase of the same in tuber extract
plus DMBA treated groups reflects the
anticarcinogenic potential and ability of
Amorphophallus Paeonifolius to revert SOD-
Catalase activity towards normalization.
Amorphophallus Paeonifolius has shown anti-
oxidant and anti-carcinogenic potential in
mammary carcinogenesis model. The results of
the present study also indicate the strong anti-
oxidant potential of Amorphophallus
Paeonifolius.

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Increase in GPX associated with of oxidative stress created by carcinogen

various forms of carcinogens has been
documented widely and the scavengers of
GPX are known to play an important role in
cancer prevention. Decrease in the level of
GPX in the liver of DMBA treated group
may be due to the decreased peroxidation of
primary substrates. GPX in Amorphophallus
Paeonifolius plus DMBA treated group
showed considerable increase towards
normalcy suggesting in protective effect of
the modulator.
GSH, acts on multiple levels of the
defense system. The thiol group of GSH
participates in the protection against
deleterious effects of reactive oxygen
species evolved during biological imbalance
as well as cancerous conditions. DMBA
treatment resulted in decreased GSH level in
the liver which is most likely a consequent
treatment and use of GSH for free radical
scavenging. The elevation of GSH level in the
Amorphophallus Paeonifolius plus DMBA
treated animals implies another protective
mechanism against electrophillic chemicals by
carcinogens.
The restoration of normal in vivo
antioxidant system and result obtained from the
effects on marmmary tumorigenesis and body
weight analysis, are the suggestive of the
potential marmmary carcinoma protective
activity of Amorphophallus Paeonifolius
against the DMBA induced mammary
carcinoma and provide further rationale for the
potential development of this trational
medicinal plant as an effective
chemopreventive agent against the mammary
carcinomas.(table 3)

Table 4: Estimation of Hematological and Bio-Chemical Parameters

DMBA + DMBA +
DMBA +
Group DMBA AP-Tuber AP-Tuber
control Tamoxifen
(n=6) only extract extract
(10mg/kg)
(50mg/kg) (100mg/kg)

WBC (x109/L) 6.95±0.13 14.09±0.14 a 7.10±0.12 a 7.08±0.22 a 7.21±0.40 a

RBC(x1012/L) 13.26±0.36 10.13±0.71 a 12.43±0.27 a 13.43±0.75 a 12.76±0.10a
Hb(g/dL) 13.26±0.36 10.13±0.71 a 12.5±0.44 b 13.43±0.75 a 12.76±0.17a
NEUTROPHIL
8.96±0.18 17.13±0.18 a 9.26±0.31 a 9.03±0.54b 8.9±0.17 a
(%)
SGPT 78.66±2.251 112.33±28.78 a 87.66±9.893c 74±7.797 a 88.66±3.141ns
SGOT 131±22.68 225.66±27.07 a 140.33±18.99 a 148.66±18.64 a 154.66±7.607a
creatinine 0.633±0.051 0.733±0.051 a 0.733±0.051 a 0.7±0.001 a 0.666±0.051 a
urea 38.56±0.459 47.5±4.588 a 30.93±4.519 a 32±2.968a 32.26±5.643 a

a-P< 0.001, b-P<0.01,c-P<0.05, ns-non significant
Data was expressed as mean ±SD. (n=6
animals in each group). Values were statistically
extremely significant at P<0.001.
The analysis of the hematological
parameters(table 4) doesn’t showed toxic
effect in rats treated with Amorphophallus
Paeonifolius alone, where as in the
experimental study, Amorphophallus
Paeonifolius extract was able to reverse the
changes in the hematological parameters
consequent to tumor inoculation. The present
study reveals that the extract was cytotoxic
towards DMBA induced tumour.
The values of RBC and Haemoglobin
were slightly lower in the DMBA treated group
than in the other experimental groups,
indicating a tendency to anemia. On the other
hand, the number of WBC was considerable
increased. The number of Neutrophils was also
significantly higher in the DMBA group,
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indicating an inflammatory response in present investigation the levels of SGPT,

animals with large tumors.
Tissue damage is the sensitive
feature in cancerous conditions. Therefore,
such a deterioration or destruction of the
membrane and lead to the leakage of
enzymes from the tissue. Hence, elevation of
these liver specific enzymes observed in
breast cancer condition and is a possible
indicator of progression of tumor growth. In
SGOT, Creatinine and Urea were found to be
increased in DMBA group. This may be due to
altered membrane permeability induced by
DMBA.
In the treatment, the levels of SGPT,
SGOT, Creatinine and Urea found to be
restored compared to DMBA treated animals
shows that extract had the optimum activity.

Table: 5 Roles of Amorphophallus Paeonifolius on the Mammary Carcinogenesis

GROUPS NO.OF RATS WITH INCIDENCES TUMOR TUMOR
TUMOR (%) LATENCY BURDEN

DMBA only 6/6 100 5±0.63 2.5±0.83

DMBA + AP-Tuber 3/6 50 5.83±0.84a 1±0.63a
extract(50mg/kg)
DMBA + AP-Tuber 2/6 33.33 6.33±1.08a 0.66±0.52 a
extract(100mg/kg)
DMBA + Tamoxifen 1/6 16.66 8.16±1.36a 0.33±0.41a
(10mg/kg)
a-P< 0.001,b-P<0.01,c-P<0.05, ns-non significant
Data was expressed as mean ±SD. (n=6 animals in each group). Values were statistically extremely
significant at P<0.001
Table: 6 Effect of Extract on The Level of Total Protein In Liver And Kidney
Experimental Rats.
DMBA DMBA
DMBA
+ +
Group DMBA +
control AP-Tuber AP-Tuber
(n=6) only Tamoxifen
extract Extract
(10mg/kg)
(50mg/kg) (100mg/kg)
0.87±0.04 0.69±0.01a 0.84±0.02 b 0.88±0.04 a 0.94±0.02 a
Kidney
0.82±0.02 0.43±0.08a 0.69±0.01a 0.78±0.02 a 0.76±0.02 a

During the 17weeks experimental The impairment is indicated by the

period the normal control rats gained in the
body weight. On the other hand, tumour
control showed no significant gain in the body
weight over the same period of time its shows
in table no 5.
The effect on tumour incidence was
also confirmed. The AP extract treatment
shows the reducing the tumour incidence
which reflects the effect of AP extract in
reduction of mammary tumour incidences
high levels of protein in tissue. In this study,
the proteins level in tumour induced animals
and AP extract treated animals had a
significant difference. This shows AP tuber
extract had the optimum activity.
Experimental and clinical study suggests
oxidative stress plays role in the pathogenesis
of a tumorigenesis. Free radicals are formed
disproportionately in tumor by non-enzymatic
glycation of proteins and subsequent

48
Research Paper
oxidative degradation of glycated protein
shown in table no 6.
4. CONCLUSION
The ethanolic extract of
Amorphophallus Paeonifolius has shown
significant antitumor and antioxidant effect in
animals. This may be due to presence of
flavanoids in ethanolic extract of
Amorphophallus Paeonifolius. The present
preliminary investigation suggests that
Amorphophallus Paeonifolius tuber stimulates
both cellular and humoral immunity. Further
studies have elucidate the exact antitumor
mechanism of Amorphophallus Paeonifolius
tuber.
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