Organic and Medicinal Chemistry

International Journal
ISSN 2474-7610

Research Article Organic & Medicinal Chem IJ

Volume 7 Issue 3 - August 2018
Copyright © All rights are reserved by Karnati Rajeswari
DOI: 10.19080/OMCIJ.2018.07.555719

Anti-Cancer Activity of Stigmasterol Of Xylocarpus

Granatum In Cytotoxicity Studies Using
Hela And Mcf-7 Cells
Karnati Rajeswari1*, T Bhaskara Rao1, SharmaGV R2 and Murali Krishna R3
1
Department of Chemistry, K L University, India
2
Department of Chemistry, Gitam University, India
3
School of Chemistry, Andhra University, India
Submission: July 25, 2018; Published: August 07, 2018
Corresponding author: Karnati Rajeswari, Department of Chemistry, KL University, Guntur, Andhra Pradesh, India,
*

Email:

Abstract

Objectives: To evaluate the anti-cancer activity of the mangrove plant Xylocarpus granatum against Hela and MCF-7 cells belonging to
identify the compound Stigmasterol unprecedented in nature by cytotoxicity analysis.

Findings: Soxhlet extraction method was used to get the corresponding extract of n-hexane. By using column chromatography extracting
solvents like n-hexane and EtOAc to get the fraction. After that fraction rechromatographed to get the compound.

Methods: The anti-cancer activities of the compound Hela and MCF-7 cells were cultivated in DMEM and ATCC out with 10% hush FBS, Penicillin,
Streptomycin in a rinse air of 5 % CO2 at 370C. Arrangement TPVG was apart with cell. Further cells are observed. After that centrifuged
50,000cells/well was seeded. At 370C, 5% CO2 hatchery. It was watched that while focus expands the restraint increments.

Improvements: At last the compound indicating great movement against Hela cells when contrast with MCF-7 cells. Docetaxel was utilized
as Standard medication.

Keywords: Mangrove Plant Xylocarpus granatum; Stigmasterol Compound; Hela and MCF-7 cells

Introduction
the in vitro confirmation of lethal impacts of obscure mixes has
Xylocarpus granatum a marine mangrove plant is a royal
been performed by numbering practical cells in the wake of
wellspring of basically one of a kind repharse and limonoids
decolouring with an essential colour. Diverse techniques utilized
as a society prescription in Southeast Asia for the operation
are estimation of radioisotope including as a measure of DNA
of looseness of the bowels, cholera, and fever-bringing on
combination, numbering via robotized counters and others
infections, for example, intestinal sickness. In the scan for
which depend on colours and cell action.
potential medication margin from tropical mangrove plants, we
have as of late announced the seclusion and different Twenty- The MTT framework is a method for evaluating the action
nine new limonoids named xylomolins A1–A7, B1–B2, C1–C2, D–F, of living cells by means of mitochondrial dehydrogenises. The
G1–G5,H–I, J1–J2, K1–K2, L1–L2, and M–N and Xylogranatins F-R. MTT strategy is straightforward, precise and yields reproducible
Examine on limonoids from the meliaceae family is of interest outcomes. The key segment is (3-[4, 5-dimethylthiazo-2-yl]-2,
because of their scope of organic exercises, for example, bug 5-diphenyl tetrazolium bromide) or MTT, is a water broken
antifeedant and evaluation controller, antibacterial, antifungal, down in tetrazolium salt passive a yellowish layout when
antimalarial, anticancer and antiviral activities. The mangroves managed in media or salt measures lacking phenol red. Refuse
xylocarpus granatum is known for creating antifeedant limonoids, MTT is remove to an insoluble purple formazan by cleavage of
particularly phragmalins and mexicanlodes. In the present paper, the tetrazolium ring by mitochondrial dehydrogenise proteins of
hostile to - malignancy movement investigations of stigmasterol proper cells. This water unsolved formazan can be solubilized
were performed which was secluded from the product of utilizing DMSO, fermented isopropanol or different solvents
X.granatum interestingly. The structure of the compound was set (Pure propanol or ethanol). The subsequent purple arrangement
up on the premise of spectroscopic information [1-5]. Actually, is spectrophotometric partner measured. An expansion or

Organic & Medicinal Chem IJ 7(4): OMCIJ.MS.ID.555719 (2018) 001

 Organic and Medicinal Chemistry International Journal

diminishing in cell number outcomes in an analogous variation in b. DMSO

the measure of formazan shaped, showing the level of cytotoxicity
c. CO2 hatchery
created by the test material. The target of the present review is
to discover the affectability of Hela and MCF-7 cell lines against d. Tecan plate peruser
Stigmasterol as watched inhibitory movement of Stigmasterol on
4.1. Predation of Test Arrangements
the expansion of both the cells.
For cytotoxicity thinks about, each test atom was weighed
Materials & Methods and blended to get the coveted focus and melted in refined DMSO
a. MTT Power (the arrangement is separated through a and volume was made up with DMEM enrich with 2% hush FBS
0.2µ m channel and put away at 2-80C for continuous utilize or to get a stock provision of 1 mg/ml fixation and soak by filtration.
solidified for amplified periods) Serial two crease weakening (0-320µg/ml) were set up from this
for completing cytotoxic reviews (Figure 1).

Figure 1: (a) MTT and (b) Formazan.

Cell lines and Culture Medium 100µl of deny test centralizations of test medications were
subsumed to the constrained monoslab in microtiter plates. The
Hela and MCF-7 Cells was trucked from ATCC, storage cells
plates were then mope at 37oC for 3 days in 5% CO2 element,
was rare in DMEM medium reinforce with 10% muffle Fetal
advance minuscule view was done; perceptions were noted for
Bovine Serum (FBS), Penicillin (100 IU/ml), streptomycin (100
each 24h. After 72h, the test groundwork in the wells was mined
µg/ml) in a stream element of 5% CO2 at 37oC in advance of
of and 50µl of MTT (5 mg/10ml of MTT in PBS) was united to
coverage. The cell was apart with TPVG layout (0.2 % trypsin,
each well. Plates were tenderly shaken and hatched for 4 h at
0.02 % EDTA, 0.05 % glucose in PBS). The suitability of the cells
37 0C in 5% CO2 air. The resilient was expelled and 100µl of
are checked and centrifuged. Further, 50,000 cells/well was
DMSO was entering and the plates were delicately overcome to
seeded in a 96 well plate and hatched for 24 hrs at 37oC, 5% CO2
solubilise the meld formazan. The absorbance was certain cover
hatchery.
a miniaturized scale plate perused at a wavelength of 540nm.
Wellspring of Reagents: DMEM, FBS, Pen strip, Trypsin The rate development restraint was ascertained utilizing the
created from Invitrogen. accompanying recipe and convergence of medication fancied to
hinder cell development by half (IC50) qualities is created from
Discussion the dosage reaction bends for every cell line.
The monoslab cell culture was trypsin zed and the cell
Results
number was traded in conformity with 1.0 x 105 cells/ml urge
DMEM involve 10% FBS. To any well of the 96 well microtiter In this review, the Compound Stigmasterol of the Mangrove
plate, 100µl of the emptied cell halt (50,000 cells/ well) was plant Xylocarpus granatum was expose to Cytotoxicity favour
admitted. After 24h a halfway monoslab was framed, the resilient utilizing MCF-7 and Hela Cells was make from ATCC, storage cells
was flicked off, scrubbed the monoslab once with medium and was refined in DMEM medium fill out with 10% hush Fetal Bovine

002 How to cite this article: Karnati Rajeswari, T Bhaskara Rao, SharmaGV R, Murali Krishna R. Anti-Cancer Activity of Stigmasterol Of Xylocarpus Granatum
In Cytotoxicity Studies Using Hela And Mcf-7 Cells. Organic & Medicinal Chem IJ. 2018; 7(4): 555719. DOI: 10.19080/OMCIJ.2018.07.555719.
 Organic and Medicinal Chemistry International Journal

Serum (FBS), Penicillin (100 IU/ml), streptomycin (100 µg/ml) underneath. The IC50 observed to be 123.2 mg/ml it has been
in a humidified status of 5% CO2 at 37oC as far as intersecting. watched that the restraint increments for that with the focus
The cell was distant with TPVG arrangement (0.2 % trypsin, increments. And after that observed to be physical portrayal are
0.02 % EDTA, 0.05 % glucose in PBS). The reasonability of the seen in Figure 3. This demonstrates advance clinical reviews are
cells are checked and centrifuged. Further, 50,000 cells/well was required. Also the compound (1) was screened against MCF-
seeded in a 96 well plate and hatched for 24 hrs. At 37oC, 5% 7 cells which have demonstrated no critical action at various
CO2 hatchery. While Decotaxel is standared sedate in this action fixations. The compound has demonstrated 28% and 34% goes
ponders. The Figure 2 was screened against Hela and MCF-7 cells. about as 160 and 320 mg/ml fixation individually. Subsequently
Strikingly, the compound has demonstrated great action against the compound is by all accounts more dynamic against Hela cells
Hela cells at various fixations as appeared in the accompanying (Figure 4).
(Tables 1 & 2). While Hela Cells Images are Figure 3 appeared on
Table 1: HELA-Cells.

S.NO Compound Name Conc. µg/ml OD at 590 nm % Inhibition IC50

1 Stigmasterol Control 0.6212 0.00

2 Stigmasterol 10 0.5983 5.05

3 Stigmasterol 20 0.5677 10.22

4 Stigmasterol 40 0.4933 22.01 123.2

5 Stigmasterol 80 0.4095 35.28

6 Stigmasterol 160 0.2906 53.96

7 Stigmasterol 320 0.2004 68.27

Table 2: MCF-7 Cells.

S.NO Compound Name Conc. µg/ml OD at 590 nm % Inhibition IC50

1 Stigmasterol Control 0.8619 0.00

2 Stigmasterol 10 0.8276 4.03

3 Stigmasterol 20 0.7955 7.09

4 Stigmasterol 40 0.7608 11.87 NA

5 Stigmasterol 80 0.6919 19.96

6 Stigmasterol 160 0.6202 28.37

7 Stigmasterol 320 0.5582 34.71

Figure 2: Chemical Structure of Stigmasterol.

How to cite this article: Karnati Rajeswari, T Bhaskara Rao, SharmaGV R, Murali Krishna R. Anti-Cancer Activity of Stigmasterol Of Xylocarpus Granatum
003 In Cytotoxicity Studies Using Hela And Mcf-7 Cells. Organic & Medicinal Chem IJ. 2018; 7(4): 555719. DOI: 10.19080/OMCIJ.2018.07.555719.
 Organic and Medicinal Chemistry International Journal

Figure 3: Cancer activity Images are observed in Hela Cells.

Figure 4: Physical Characteristic of Stigmasterol.

Conclusion References
We have communicated in this paper the results of the 1. Zhen Fang Z, Tiber K, Attila M, Li Gong Y, Yu Cheng G, et al. (2016) Novel
and Neuroprotective Tetranortriterpenoids from Chinese mangrove
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of X. granatum using Hela and MCF-7 cells. It was found
2. Xi Yin, Xin Li, Yaoguang H, Yiwen Z, Haishui S (2015) Xylocarpin H, a
that the compound showing good activity against Hela cells Limonoid of Xylocarpus granatum, Produces Antidepressant Activity.
when compared to MCF-7 cells. Form the present study, it has Journal of Behavioral and Brain Science. 5: 524-532.
been concluded that the compound is showing good anti- 3. Zhen Fang Z, Orazio Taglialatela S, Yu Cheng Gu, Ling Yi K (2014)
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communicated. granatum. Fitoterapia 97: 192-197.
4. Wenping C, Li Shen, Tirumani Satyanandamurty, Jun Wu (2014)
Acknowledgment Absolute configurations of new limonoids from Xylocarpus granatum.
Fitoterapia 94: 108-113.
We are grateful to Department of Chemistry K L University,
Andhra Pradesh, India acknowledged for necessary facilities. 5. Yibing Wu, Ying Bai, Xiaohan Guo, Jinlong Qi, Mei Dong, et al. (2014)
A new Limonoid from Xylocarpus granatum. Chemistry of Natural
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004 How to cite this article: Karnati Rajeswari, T Bhaskara Rao, SharmaGV R, Murali Krishna R. Anti-Cancer Activity of Stigmasterol Of Xylocarpus Granatum
In Cytotoxicity Studies Using Hela And Mcf-7 Cells. Organic & Medicinal Chem IJ. 2018; 7(4): 555719. DOI: 10.19080/OMCIJ.2018.07.555719.
 Organic and Medicinal Chemistry International Journal

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How to cite this article: Karnati Rajeswari, T Bhaskara Rao, SharmaGV R, Murali Krishna R. Anti-Cancer Activity of Stigmasterol Of Xylocarpus Granatum
005 In Cytotoxicity Studies Using Hela And Mcf-7 Cells. Organic & Medicinal Chem IJ. 2018; 7(4): 555719. DOI: 10.19080/OMCIJ.2018.07.555719.