International Journal of Civil Engineering and Technology (IJCIET)

Volume 8, Issue 1, January 2017, pp. 943947 Article ID: IJCIET_08_01_111

Available online at http://www.iaeme.com/IJCIET/issues.asp?JType=IJCIET&VType=8&IType=1
ISSN Print: 0976-6308 and ISSN Online: 0976-6316

IAEME Publication Scopus Indexed

ANTI-CANCER ACTIVITY OF PHYLLANTHUS

RETICULATUS ON COLON CANCER CELL LINE
C. Aarthi
Research Scholar, Department of Industrial Biotechnology
Bharath University, Chennai, India

Dr. P.B. Ramesh Babu

Professor & Head, Department of Genetic Engineering,
Bharath University, Chennai, India

ABSTRACT
Cancer is a global epidemic that is still a mystery in the field of medicine. Phyllanthus
species is proven to have various medicinal and thereuptic properties. In this study we have
tried to explore the anti-cancer properties of Phyllanthusreticulates using MIT assay in -
vitro.colon cancer cell lines -HT 29 purchased from Pune was used for study.
Key words: Cancer, Phyllanthu, global epidemic, Anticancer
Cite this Article: C. Aarthi and Dr. P.B. Ramesh Babu, Anti-Cancer Activity of Phyllanthus
Reticulatus on Colon Cancer Cell Line. International Journal of Civil Engineering and
Technology, 8(1), 2017, pp. 943947.
http://www.iaeme.com/IJCIET/issues.asp?JType=IJCIET&VType=8&IType=1

1. INTRODUCTION
Cancer is a global epidemic with approximately fourteen million new cases being diagnosed each
year, leading to an annual death toll of approximately eight million (Ferlay et al., 2015).
Herbs and plants derived medicines have a long history of use in the various treatments and now,
they still remain as important sources for the development of anticancer drugs. More than 3000 plants
species have been reported to be involved in the development of anticancer drugs (Shoeb, 2006). From
1940-2006, more that 40% of drugs in the market are anticancer agents and 65% of these anticancer
drugs mimic natural compounds.
Phyllanthus (EUPHORBIACEAE) is a small annual plant and is widely distributed throughout the
tropical and subtropical regions of the world (Lee et al., 1996).This genus consisting of more than 700
species. Phyllanthus has been employed as herbal drugs for a long time in China, India, Brazil, and
Southeast Asian countries. These plants have bitter and astringent taste and are considered as
stomachic, diuretic,febrifuge, deobstruent, antiseptic, and effective remedies for hepatopathy. In
India, fifteen species of genus Phyllanthus are widely used by indigenous medicine.
The extracts of these plants have been reported to have pharmacological effects such as antiviral
activity against hepatitis B and related hepatitis viruses,anti-bacterial activity, anti-hepatotoxic or
liver-protecting activity and hypoglycaemia properties (Etta, 2008).

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 Anti-Cancer Activity of Phyllanthus Reticulatus on Colon Cancer Cell Line

The anticancer effects of the genus Phyllanthus has been reported in a few papers, for instance P.
amarus protects the liver from hepatocarcinogenesis and the root extract of P. acuminatus exerts
growth inhibition in murine P-388 lymphocytic leukaemia and B-16 melanoma cell lines (Pettit et al.,
1990; Powis & Moore, 1985). Reports on the anticancer activity of P.reticulatus are meagre. To throw
more light on this aspect the present study on the anticancer activity of P.reticulatus against colon
cancer cell line was planned and carried out.

2. MATERIALS AND METHODS

The aerial parts (leaves and fruits) of Phyllanthus reticulates were collected from Chennai, Tamil
Nadu and were shade dried, powdered and stored in an air container. 25 grams of power was dipped
into 250 ml of Ethanol and water separately and kept inside the rotary shaker for 7 days. After 7 days
the extracts were filtered through Teflon cloth and whatman No.1 filter paper. The filtrates were
concentrated using rotary evaporator. After evaporation stock solutions were prepared to a final
concentration of 100mg/ml.

2.1. Anti Cancer Study

COLON CANCER cell line HT29 was purchased from NCCS, Pune. The cells were maintained in
Minimum Essential medium supplemented with 10 % FCS, Penicillin (100 U/ml) and Streptomycin
(100ug/ml) in a humified atmosphere of 5% Co2 at 370C.

2.2. In Vitro assay for Anticancer Activity (MTT assay) (Mosmann, 1983)
Cells (1 105/well) were plated in 24-well plates and incubated in 370C with 5% CO2 condition. After
the cell reaches the confluence, the various concentrations of the samples were added and incubated
for 24hrs. After incubation, the sample was removed from the well and washed with phosphate-
buffered saline (pH 7.4) or MEM without serum. 100l/well (5mg/ml) of 0.5% 3-(4,5-dimethyl-2-
thiazolyl)-2,5-diphenyl--tetrazolium bromide (MTT) was added and incubated for 4 hours. After
incubation, 1ml of DMSO was added in all the wells .The absorbance at 570nm was measured with
UV- Spectrophotometer using DMSO as the blank. Measurements were performed and the
concentration required for a 50% inhibition (IC50) was determined graphically. The % cell viability
was calculated using the following formula:
% cell viability = A570 of treated cells / A570 of control cells 100
Graphs are plotted using the % of Cell Viability at Y-axis and concentration of the sample in X-
axis. Cell control and sample control is included in each assay to compare the full cell viability
assessments.

3. RESULTS AND DISCUSSION

The percentage cell viability of ethanol and aqueous extracts were shown in table 1 and 2.The IC50
values of aqueous and ethanol extracts of Phyllanthus reticulates was recorded at15.6ug/ml and
31.2ug/ml respectively. The results clearly showed that the extract of Phyllanthus reticulatus were
found to be effective to suppress the growth of colon cancer cell line-HT29.Further the results confirm
that aqueous extract was more effective than Ethanol extract against the colon cancer cell lines.
Sumalatha (2013) reported the anticancer activity of Phyllanthus emblica against colon cancer cell
line. The results clearly exhibit that the Phyllanthus species are potential agents to be utilized as an
anticancer drugs. Further studies are warranted on these plants to examine the potential of these plants
to prevent the cancers of diverse tissue sites and the active principle of these plants.

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 C. Aarthi and Dr. P.B. Ramesh Babu

Table 1 Anticancer effect of aqueous extract on HT29 cell line

S.No Concentration Dilutions Absorbance Cell Viability
(g/ml) (O.D) (%)
1 1000 Neat 0.032 1.82
2 500 1:1 0.120 6.84
3 250 1:2 0.298 16.98
4 125 1:4 0.386 22.00
5 62.5 1:8 0.499 28.44
6 31.2 1:16 0.609 34.72
7 15.6 1:32 0.872 49.71
8 7.8 1:64 1.019 58.09
9 Cell control - 1.754 100

Cell Viability (%)

120
P
e100
r
80
c
e 60
n
t 40
a
g 20
e
0
1000 500 250 125 62.5 31.2 15.6 7.8 Cell
Concentration(g/ml) control

Figure 1 Cell Viability-Percentage

Table 2 Anticancer effect of Ethanol extract on HT29 cell line

S.No Concentration Dilutions Absorbance Cell Viability
(g/ml) (O.D) (%)
1 1000 Neat 0.051 2.90
2 500 1:1 0.138 7.86
3 250 1:2 0.228 12.99
4 125 1:4 0.332 18.92
5 62.5 1:8 0.493 28.10
6 31.2 1:16 0.883 50.34
7 15.6 1:32 1.043 59.46
8 7.8 1:64 1.236 70.46
9 Cell control - 1.754 100

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 Anti-Cancer Activity of Phyllanthus Reticulatus on Colon Cancer Cell Line

Cell Viability (%)

120
P100
e
r 80
c
60
e
n 40
t
a 20
g 0
e 1000 500 250 125 62.5 31.2 15.6 7.8 Cell
control
Concentration(g/ml)

Figure 2 Cell Viability-Percentage

4. CONCLUSION
The anti-cancer effect study of Phyllanthus reticulates on colon cancer cell line-HT 29 using MIT
assay is found to be commendable. Aqueous extract of Phyllanthusreticulates shows 50% cell viability
at 15.6 milligram/ ml, whereas the ethanol extract shows 31.2 milligram/ ml. Further research on the
extracts might be useful in finding effective drugs in the treatment of cancer.

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