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PHYTOCHEMICAL ANALYSIS OF THE LEAF OF Dieffenbachia maculate
Schott (DUMB CANE) FOR CYTOTOXICITY ASSAY AND
ANTIANGIOGENIC TESTING USING CHORIOALLANTOIC
MEMBRANE (CAM) OF DUCK EMBRYOS

A Research Proposal Presented to the

Biological Sciences Department
College of Science and Computer Studies
De La Salle University-Dasmarias
City of Dasmarias, Cavite

In Partial Fulfilment of the Requirements for the Degree

Bachelor of Science in Medical Biology

SUHAIMA N. DALIMBANG
JANINA SUZETTE C. GUILLERMO

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May 2015
CHAPTER 1
INTRODUCTION

1.1

Background of the Study

People can safely say that the medical field, in this modern time, is already

developed, advanced and well established. Concurrently, we can expect that

modernization has helped science understand and treat most diseases and ailments
but despite the progress and growth of our knowledge there are still sicknesses
that are not fully understood. One disease that until now has no treatment was
discovered in Egypt and dates back to about 3000 BC called Edwin Smith
Papyrus or Cancer, in todays term (Cancer Organization c2005-2015).
According to American Cancer Society, cancer is the uncontrolled growth
of abnormal cell, called malignant cells, in the body. Treatment of cancer includes
the use of chemotherapeutic drugs, radiation therapy or surgery; all these are
available to help lengthen the life of a patient but none of them can truly eradicate
and cure cancer Due to this, World Health Organizations statistics showed that
there were 14.1 million new cancer cases, 8.2 million cancer death and 36.2
million people living with cancer in 2012 worldwide (WHO 2012). Unfortunately,
based on Department of Health, cancer ranked third in the list of leading causes of
in the Philippines. Also, it has been estimated that about 18.6 persons per 100,000

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Filipinos now living will eventually have cancer, one every five (5) Filipinos who
live to age 74 will get cancer (DOH 2010). This phenomenon is still a challenge
to scientists; certain studies are being conducted to create a medicine that will
cure cancer because the treatments that are available at this moment are very
expensive and not affordable to most people.
Accordingly, the need for a safe and reliable treatment and drug is urgent.
According to Lewin Jo (2014) a natural therapist, plants and herbs have been used
by herbalists and apothecaries throughout the centuries because of their healing
and restorative powers. Plants contains phytochemical constituents that has the
potential in treating diseases like cancer, heart disease, pneumonia, diabetes, etc.
According to Dr. Winston Craig, certain plants contain flavonoids, which help
vitamin C work more efficiently as an antioxidant, mopping up the free radicals
that cause cancer. Some plants also contain terpenoid, which is thought to inhibit
growth tumor (Jo 2014). A plant that has the potential to treat cancer is
Dieffenbachia seguine, commonly known as Dumb Cane Plant. This plant is used
as a toxin to poisonous arrows and as a gardening plant for ornamental purposes.
This plant was discovered by researchers in Notre Dame Dadiangas University in
General Santos City to be useful in controlling the spread of cancer cells in the
body because it contains antiangiogenic factors that inhibit the growth and
development of new blood vessels. They used wild duck embryos and observed
the effects of dumb cane essential oil, leaf extracts, mineral water and

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Simvastatin. Results showed that there was a lower blood vessel braching point
compared to embryos in mineral water. Thus, this plant has a great potential in in
the formulation of anti-cancer drug.
This study will determine the specific phytochemical components of dumb
cane plant and its antiangiogenic effect to chorioallantoic memberane.

1.2

Conceptual Framework
According to American Cancer Society (2009), Anti-angiogenesis is a

form of targeted therapy that uses drugs or other substances to stop tumors from
making new blood vessels. Without a blood supply, tumors can't grow. Recent
studies has hypothesized that Dieffenbachia seguine has the ability to inhibit
blood vessel formation to stop the growth of tumor cells. The paradigm of the
proposed study is given below:

A: Plant Extract containing

Phytochemical Constituents
of Dieffenbachia maculata
B: Mineral Water
C: Simvastatin

Blood Vessel Formation of

Duck Embryo

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This study was conducted to determine the Plant Extract containing

Phytochemical Constituents of Dieffenbachia maculata, Mineral Water and
Simvastatin will be introduced to 10 days old duck embryo for observance of
blood vessel formation. The plant extract would show the antiangiogenic effect,
the mineral water would show the normal growth of blood vessels while the
simvastatin would show the angiogenic effect on the embryo. If the plant extract
has a positive result for antiangiogenesis, it will be used to test its ability to inhibit
tumor growth.

1.3

Statement of the Problem

This study generally aims to determine the Phytochemical analysis of the

leaf of Dieffenbachia maculata (Dumb cane) for cytotoxicity assay and

antiangiogenic activity testing using Choriallantoic Membrane (CAM) of duck
embryo.
Specifically, the study seeks to answer the following questions:
1. What are the phytochemicals present in the leaf of Dieffenbachia
maculata?

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2. Do the following compounds subjected to Chorioallantoic Membrane
(CAM) test using duck embryos will result positive for antiangiogenic
properties?
3. What is the population of viable cells after application for assessing the
cytotoxicity assay of Dieffenbachia maculate?

1.4

Scope and Limitations

This study focused on the extraction of phytochemicals on the leaf of

Dieffenbachia maculate (dumb cane) and isolation of active compounds with

antiangiogenesis efficacy. The extraction preparation of phytochemicals only used
the solvent extraction method.
Determining the antiangiogenic properties and cytotoxicity of the plant
extract are the evaluation scope of this study after phytochemical analysis. Duck
embryo is used in Chorioallantoic membrane (CAM) which will be tested for 10
days and compounds such as mineral water (phosphate buffer saline solution) and
simvastin, a commercial anti-cholesterol drug were also used to determine if the
plant extract effect will be comparable to these compounds effect.

1.5

Significance of the Study

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This study aims to help the progress in research of cancer treatment with
the use of herbal medicine. The potential therapeutic claims of Dieffenbachia
maculata will be analyze through phytochemical analysis and antiangiogenesis.
The result of this study will be beneficial and helpful to the following:
World Health Organization and Cancer Organizations. This would be of
great significance to those organizations looking for a treatment to cancer because
this research could be a step in finding cure to cancer by using herbal plant as a
therapeutic drug.
Department of Health. There is a possibility that a drug can be
manufactured with the help of this study because of its availability in the
Philippines. It would give recognition to our country and would lessen the
mortality rate of Filipinos.
Pharmaceutical researchers. This study can be used to update treatment
methodologies and guide to them in producing a commercially available anticancer drug.
Cancer Patients. The possibility of manufacturing a drug will be great
because this plant is common and very affordable. All cancer patients, regardless
of their social and economic capability, will have the access to a safe and effective
treatment.

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Further Researchers. This research investigation may serve as baseline
information and can be used as a reference for them to conduct similar research
study in the future.

1.6

Definition of Terms
The following statements define terms within the perspective of the study:
Antiangiogenesis. A process that inhibits the growth or formation of new

blood vessels within the present tumor to prevent metastasis.

Chorioallantoic

membrane. A higly vascularized

extra-embryonic

membrane formed in birds and reptiles by the apposition of the allantois to the
inner surface of chorion.
Cytotoxicity. It is the quality of being toxic to cells that causes them to
undergo apoptosis or programmed cell death.
Dieffenbachia maculata. Commonly known as Dumb cane. It posses
phytochemicals with antioangiogenic properties extracted from its leaf.
Metastasis. The process of spreading the diease producing agency as
cancer cells from the initial primary site of disease to another part of the body.

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Phytochemicals. A natural occurring bioactive compounds in plants that
are considered beneficial for human health.
Simvastatin. A compound commercially available in the market that acts
by inhibiting cholesterol synthesis.
Tumors. An abnormal growth of tissue resulting from uncontrolled,
progressive multiplication of cells serving no physiological functions.
Viable cells. A number of cells that is alive and capable of growth and
proceeding to its normal function as cell.

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CITED REFERENCES

(1) Bashir Arif, Saxena Trisha. 2013 Evaluation of Antiangogenic and

Antiproliferative Efficacy of 3 Phytochemicals with Special reference to
Anthocyanins. International Journal of Science and Research (IJSR), Bhopal
Memorial Hospital and Research centre.
(2) Cancer Organization [Internet]. c2005- 2015. American Cancer Society,
Incorporated: [cited: 2015 May 15]. Available from: http://www.cancer.org/

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(3) Department of Health [Internet]. c2011. Health Statistics: Leading Cause of
Mortality.

[cited:

2015

May

15].

Available

from:

http://www.doh.gov.ph/statistics.html
(4) Jeong, Koh, Lee, Bae, Lu, Kim (2010). Review of Antiangiogenic
Phytochemicals and Medicinal Herbs, College of Oriental Medicine, Kyung Hee
University Seoul 130-701, Republic of Korea. Published online DOI:
10.1002/ptr.3224: www.interscience.wiley.com
(5) Lewin Jo [Internet]. c2015. BBC Good Food: The healing Power of Herbal
Medicine.

[cited:

2015

May

15].

Available

from:

http://www.bbcgoodfood.com/howto/guide/healing-power-herbs
(6) Philippine Council for Health Research and Development (2013). Dumb cane
plant found potential in treating cancer. [cited May 28, 2013]; 2723 Available
from:

http://www.pchrd.dost.gov.ph/index.php/news/2723-dumb-cane-plant-

shows-potential-in-treating-cancer
(7) World Health Organization [Internet]. c2015. Cancer: Country Profiles.
[cited: 2015 May 15]. Available from: http://www.who.int/cancer/en/

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APPENDIX A
CURRICULUM VITAE

Suhaima N. Dalimbang

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Suhaima N. Dalimbang was born on September 16, 1997 in Makati City,
Metro Manila. With her supportive parents Hassan Dalimbang and Zenaida
Dalimbang, she started her grade school years in Philippine Womens
University and graduated from secondary school at St. Scholasticas college
Manila with excellence in academic award. She showed her interest in the
filed of science especially in Biology. She is pursuing her tertiary education
at De La Salle University Dasmarias and took the course Bachelor of
Science in Biology Major in Medical biology. Suhaima, with her undying
passion in the field of science plans to pursue her degree in Medicine as a
cardiologist with a purpose of giving important contributions in the field of
Medicine and her country, Philippines.

Janina Suzette C. Guillermo

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Janina Suzette C. Guillermo was born on February 21, 1997 in University of Sto.
Tomas Hospital, Manila. With her supportive parents, Matias Jun V. Guillermo
and Susana C. Guillermo, she started her grade school years and also graduated
from secondary school in Lords Angels Montessori School both as Valedictorian
of her batch. Being a consistent honor student, she showed her interest in the field
of Science especially in Biology. She is pursuing her tertiary education at De La
Salle University Dasrmarias and took the course Bachelor of Science in
Biology Major in Medical Biology. With her undying passion and dream of
becoming a Medical doctor, Janina is planning to continue her aspirations in the
Medical school and further apply for specialization as a Cardiothoracic Surgeon.
She believes that being a doctor is not about gaining money, fame, title or just a
degree but she has visualized this career as calling for some people in order to
help and prolong lives.

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