Anti cancer thesis 1

Anti Cancer Activity of
Medicinal Plant
On breast cancer or blood cancer
MBT Project Thesis-Part 1
Guided by Prepared by
Dr. Rabiul Haque,Ph.D Sk Abu Raihan Abdullah
Professor & HOD MBT , 3rd
Semester
Department of Biotechnolgy NITMAS
NITMAS

INTRODUCTION
I. Overview
Cancer is one of the most serious health problems worldwide, affecting
individuals from different sexes, ages, and races. It is a group of diseases,
characterized by uncontrolled cellular growth with frequent cancer cells
invasion to different body parts and spreading to other organs,
a process referred to as Metastasis. Metastasis is the major cause
of cancer related mortality.
In 2005, cancer was the second leading cause of death among both men and
women and accounted for 13% of the total 58 million deaths worldwide . In
2006, about 10.9 million new cancer cases are expected to be diagnosed
worldwide and more than 7.8 million cancer patients may die .
According to the latest report of cancer registry unit in Gaza strip, 5500 cases
have been reported over the period from January, 1995 to December, 2003 .
In addition, 1026 cancer patients died in 2004 in the Palestinian territories with
a mortality rate of 28.2 per 100.000 .
Cancer is also a problem of economical dimensions with a very high level of
expenses associated to it. For example the National Institute of Health, USA
estimates that an overall of $209.9 billion were invested worldwide in 2005,
for the sake of cancer research and management .
Cancer is a heterogeneous illness which can originate from many different
organs of the human body. However, the most frequent cancer types in the
world are lung, prostate, stomach, colorectal, and esophagus in men; and
breast, lung, stomach, colorectal and cervical in women .
Prostate cancer is the most frequently diagnosed and the second leading cause
of cancer death among men, with 234460 new cases estimated to occur in USA
during 2006, and 27350 American men will die as a result of this disease(3). In
Palestine, the mortality rate of prostate cancer was 1.4 per 100000 during the
period from January, 1995 to December, 2002 . Despite the fact there are
several cell types in the prostate, nearly all of the prostate cancers are
adenocarcinoma, originating in the gland cells .

Liver cancer ranks as the sixth most common type of cancer world wide .Many
different liver related tumors are identified depending on the type of cells
where they originate, from these types about 83% are hepatocellular
carcinoma (HCC) that begin in the hepatocytes, the main type of liver cells.
Cervical cancer is the most common cause of cancer death among women in
developing countries and the second most common caner in women
worldwide. It is caused by a change in the epithelial cells, which
line the wall of the cervix, and the most common risk factor for this type
of cancer is the human Papillomavirus (HPV).
In the last decades there were great advances in the diagnosis of cancer as well
as in the field of molecular oncology. However, the cure rate of most cancers
remains low. Several strategies have been used to cure cancer among which
the most common are surgery, chemotherapy, radiotherapy, and
immunotherapy. Other modern approaches such as hormonal and gene
therapy were proposed by researchers to replace conventional cancer therapy,
with variable degrees of success .
All of these therapies have undesired side effects, they are usually not
available all the time and they are expensive. For instance, in surgery
the immune system is compromised due to the large amount of cortisole
released subsequent to the surgery, which increase the probability of cancer
relapse .
Moreover, the current use of chemotherapy is accompanied with
difficult side effects. It inhibits bone marrow stem cells proliferation leading to
immune suppression . Radiotherapy which is widely used in the world is also
accompanied by a great deal of side effects. Lymphocytes are most readily
affected by radiation resulting in prolonged T-cell suppression . Other side
effects such as, bone necrosis, lung fibrosis, skin devascularization, ulceration,
nausea, vomiting, and renal damage are also associated with all types of
conventional therapies.
As the conventional cancer therapies failed to completely fulfill the criteria for
a successful cancer therapy, the use of naturally developed anticancer agents
has evolved as an alternative safe, low-cost and convenient one. Nontoxic
chemoprevention agents from natural resources were proposed by
researchers for this purpose.

II. Objectives
The major scientific and technological progress in combinatorial chemistry,
drugs derived from natural products still make an enormous contribution to
drug discovery today. Efforts therefore are being made to identify naturally
occurring anti-carcinogenesis which would prevent, slow and/or reverse
the cancer induction and its subsequent development. Plants are good
reducing factors for cancer. This thesis deals with the anti-cancer benefit
of drug isolated from plant.
The aim of the current study is to investigate the effect of Solanum
viarum(Tropical Soda Apple), Acanthus illicifolious (Harkucha Kanta), Annona
squamosa(Custard Apple), Alstonia scholaris(Chatium) and Calotropis gigantea
(Akanda) leaf extracts on human breast cancer or blood cancer.
Specifically this study aims at:
1- Determination of the proliferation activities of each cell line in response to
each plant extract treatment.
2- Determination of percent viability of each cell line in response to each
plant extracts treatment.
3- Determination of any morphological changes of each cell line in each time
performing viability testing assay in parallel.

LITERATURE REVIEW
I. Conventional cancer therapy
Despite the latest advances in medical sciences, and progress in strategies of
cancer treatment, cancer currently remains a tragic disease and is one of the
major causes of death worldwide. The principal methods of cancer treatment
include chemotherapy radiotherapy and surgery.
Chemotherapy is a systemic treatment, to which the whole body is exposed.
Among the most successful chemotherapeutic agent are Cisplatin, Mitomycin
and Docetaxel. All of these agents enhance serious side effects or long term
complication . These side effects include kidney damage, hearing
loose, lower blood count, liver damage, nerve damage, and blood vessel
damage.
External beams of Radiotherapy are associated with unacceptably high levels
of local-regional toxicity . Particularly, it affects the rapidly dividing cells of
mucosa, causing irritative urinary and blood loss . Later toxic effects result
from damage to the more slowly proliferating cells such as fibroblasts,
endothelial, or paranchymal stem cells causing chronic fibrosis and vascular
damage .
Other undesired side effects, such as, immune suppression, bone necrosis,
lung fibrosis and skin devascularization are seen with all types of
conventional therapies .

II. Plants with anticancer effects
Plants are the chief source of natural products that are used in medicine. Even
Aspirin, the world best known and most universally used medication, has its
natural origins from the glycoside salicin which is found in many species of the
plant genera Salix and Populus .
The scientific literature is rich in epidemiological studies that support
significant differences in the occurrence of cancers between oriental
and occidental populations . Generally, populations that consume
a high level of natural herbal products have a reduced incidence of cancer.
An example is the low incidence of colon cancer in Asian countries with high
consumption of soybean products . Soy beans are the major dietary source of
saponins, which have been suggested as possible anticancer agents .
There is lately a great interest in screening for plants to be used in cancer
prevention and treatment. For this reason,leaf extracts from different plants
have been extensively studied and confirmed anticancer activity.

Studies that examined the therapeutic
potential of plants leaf extracts
Plant Scientific Name Common Name Type of extraction Proposed active material
1. Solanum viarum Tropical Soda Apple Ether Solasodine glycoalkaloid
2. Acanthus illicifolious Harkucha Kanta Methanol Titerpenoids,Flavonoids,
Alkaloids
3. Annona squamosa Custard Apple Ethyl acetate Acetogenins,Alkaloids,
Dofamine
4 Alstonia scholaris. Chatium Methanol Alkaloids,Flavonoids
5. Calotropis gigantea Akanda Ethanol Triterpenoids,Flavonol
Glycosides

Selected Plants for leaf extract
Scientific Name : Solanum viarum Dunal
Common Name : Tropical Soda Apple
Solanum viarum Dunal, belongs to the family Solanaceae, commonly called
Tropical Soda Apple (TSA) is a shallow rooted, profusely branched, perennial
shrub. It is native plant of Argentina and central Brazil (Mullahey et al, 1993)
and has been naturalized in many parts of the world including India. Solasodine
is a major glycoalkaloid constituent of this plant and it is a as the potential
alternative for diosgenin which share a characteristic conversion to 16-dehydro
pregnenolone acetate, the first step in steroid synthesis (Bhudavari, 1989).
Solasodine extracted from this plant is used as a steroid precursor to produce
contraceptive pills (Everist, 1981), treat cancer (Trouillas et al., 2005),
Addison's disease, rheumatic arthritis, chronic asthma, leukemia (Blood
cancer), obesity, palsy, skin diseases (Pingle and Dhyansagar, 1980), and anti-
inflammatory steroidal drugs
(Pandurangan et al., 2011).
Tropical soda apple plant(Solanum viarum)

Plant leafs Collection,Identification and extraction
The Tropical soda apple plant leafs was collected from Haldia river side area in
August,2015 and identified Botanical Survey of India,Botanical
Garden,Howrah-711103
Air dried plant leafs (100gm) were powdered in a mechanical grinder.
And the plant leafs powder were successively extracted by aquaous,
ether,ethanol,ethyl acetate, chloroform and methanol by using soxhlet
extraction apparatus. Used 1 gm powder used for each exraction.
Then solvent was completely removed under reduced pressure and stored in
a vacuum dessicator. The yields of the ether,ethanol, chloroform and methanol
fraction were about 7.25% and 9.30 % respectively.
Qualitative analysis
Preliminary qualitative analysis has been needed performed to know the
type of compounds present in the various extracts
Further needed chemical group tests were performed for identified
salasodine glycoalkaloid , others and the yields of these ether,ethanol,
chloroform and methanol extracts were noted. These important contains
are functioning anti cancer activity in cancerous cell line.

Scientific Name : Acanthus ilicifolius Linn.
Common Name : Harkucha Kanta
Acanthus ilicifolius Linn. (Acanthaceae) a gregarious shrub, commonly found
with mangroves along the coastal regions of India. The common names of
Acanthus ilicifolius are Beng. Harkucha kanta, Eng. Holy leaved acanthus.
It is found commonly in the east and west coasts, also found in Meghalaya and
the Andamans. Leaves are simple, oblong to elliptic, varying in size, smaller in
flowering season.
Acanthus ilicifolius (Acanthaceae) has received considerable attention due to its
wide range of secondary metabolites and its traditional usage in Indian and
Chinese system of medicine. This plant is reported to be a mangrove.Mangrove
survives in the most hostile environment with fluctuating tidal and saline
regime. Hence, these plants are considered to be rich sources of
steroids,triterpenoids, saponins, flavonoids, alkaloids, and tannins.
The plant have demonstrated significant pharmacological activities like
antioxidant, anticarcinogenic, anti-osteporotic and hepatoprotective.
Traditionally, the plant has been used for dyspepsia, paralysis, asthsma,
headache, rheumatism, and skin diseases.
Harkuncha kanta Shrub(Acanthus ilicifolius)

Plant leaf Collection,Identification and extraction
The Harkucha Kanta Shrub leafs was collected from Haldia river side area in
And the plant leafs were successively extracted by aquaous,
extraction apparatus. Used 1 gm powder used for each exraction.
compounds like steroids,triterpenoids, saponins, flavonoids, alkaloids, and
tannins. of these ether,ethanol, chloroform and methanol extracts were
noted.These important contains are functioning anti cancer activity in
cancerous cell line.

Scientific Name : Annona squamosa Linn.
Common Name : Custard apple
Annona squamosa Linn. (Annonaceae), popular with common name of
“custurd apple or sugar apple”.
Its origin from West Indies and Central America. The plant is also cultivated
throughout the South Asia especially in tropical regions for edible purpose. It is
a small (about 5-6 m in height) deciduous tree with irregular branches.
Ethyl acetate extracts leaf of Annona squamosa L. were found
acetogenins,alkaloids,dofamine,salsolinol and Coclaurine. Its have significant
anticancer activity. It is effective remedy for various types of inflammatory
diseases as well as tumorous growths.
The plant has been used in traditional medicine of several tropical countries to
treat epilepsy, dysentery, cardiac problems, worm infection, constipation,
hemorrhage, antibacterial, dysurea, fever and ulcer. It also has anti-fertility
and abortifacient properties (Soni et al., 2012).
Custard apple tree(Annona squamosa Linn.)

The Custard apple tree leafs was collected from Haldia river side area in
extraction apparatus.
type of compounds present in the various extracts.
compounds like acetogenins, alkaloids, dofamine, salsolinol and Coclaurine
of these ether, ethanol, chloroform and methanol extracts were noted.
These important contains are functioning anti cancer activity in cancerous
cell line.

Scientific Name : Alstonia scholaris Linn.
Common Name : Saptaparni or Chatium
Alstonia scholaris Linn. (Family Apocynaceae) which is popularly known as the
“Saptaparni” or “Devil’s tree”. Common name of Alstonia scholaris Linn. are
Bengali Chatium.
Alstonia scholaris Linn. is a medium to large tree. The upperside of the leaves
are glossy, while the underside is Greyish, its mature bark is grayish.
It is widely distributed in dried forests of India, Westwern Himalayas,Western
Ghats and in the Southern region.
The plant Alstonia scholaris has been used in different system of traditional
medication for the treatment of diseases and ailments of human beings. It is
reported to contain various alkaloids, flavonoids and phenolic acids. It has
been reported as anti-cancer, antioxidant, analgesic, anti-inflammatory, anti-
ulcer, bronchodilatory, antimicrobial, antiamoebic, antidiarrhoeal,
antiplasmodial, hepatoprotective, immunomodulatory, antiasthmatic, free
radical scavenging, anti-fertility and wound healing activities.
Saptaparni Chatium tree(Alstonia scholaris Linn.)

The Saptaparno or Chatium tree leafs was collected from Haldia river side area
in August,2015 and identified Botanical Survey of India,Botanical
compounds like alkaloids, flavonoids and phenolic acids of these
ether,ethanol, chloroform and methanol extracts were noted.These
important contains are functioning anti cancer activity in cancerous cell
line.

Scientific Name : Calotropis gigantea
Common Name : Akanda
Calotropis gigantea (Crown flower or Akanda) is a species of Calotropis native
to Cambodia, Indonesia, Malaysia, Philippines, Thailand, Sri Lanka, India, China,
Pakistan, Nepal, and tropical Africa.
It is a large shrub growing to 4 m (13 ft) tall. It has clusters of waxy flowers that
are either white or lavender in colour. The plant has oval, light green leaves
and milky stem.
Several phytochemicals have been isolated from Calotropis gigantea and they
include cytotoxic cardenolides, antifeedant nonprotein amino acid,
naphthaleneand terpene derivatives, flavonol glycosides, pregnanes, ursane-
type triterpenoids and steroids.
Traditionally extracts and preparations from roots and leaves of Calotropis
gigantea are used against abdominal tumours, boils, syphilis, tuberculous,
leprosy, skin diseases, piles, wounds, rheumatism and insect-bites.
Root bark of this plant is used in dysentery and as a purgative,
alterative,diaphoretic and emetic. Antipyretic, analgesic, anticonvulsant,
anxiolytic, sedative, hepatoprotective, wound healing, antidiabetic,
larvicidal,anti-inflammatory and anti-diarrhoeal activities of C. gigantea have
been scientifically proven.

Akanda shrub (Calotropis gigantean)
The Akanda shrub leafs was collected from Haldia river side area in
compounds like flavonol glycosides, pregnanes, triterpenoids and steroids of
these ether,ethanol, chloroform and methanol extracts were noted.These
important contains are functioning anti cancer activity in cancerous cell
line.

References
1. Chandra, V., and S. N. Srivastava. 1978. Solanum viarum Dunal
syn. Solanum khasianum Clarke, a crop for production of
solasadine. Indian Drug. 16: 53-60.
2. Nee, M. 1991. Synopsis of Solanum section Acanthophora: A group
of interest for glyco-alkaloides, pp. 258-266 In: J. G. Hawkes, R. N.
Lester, M. Nee and N. Estrada eds. Solanaceae III: Taxonomy,
chemistry, evolution. Royal Botanic Gardens Kew, Richmond,
Surrey, UK.
3. Khajure, P. V. & Rathod, J. L. (2011). Potential anticancer activity of
Acanthus ilicifolius extracted from the mangroves forest of Karwar,
west coast of India. World Journal of Science and Technology, 1(1),
1-6.
4. Wostmann, R. & Liebezeit, G. (2008). Chemical composition of the
mangrove holly Acanthus ilicifolius (Acanthaceae). Senckenbergiana
maritime, 38(1), 31-37
5. Neha Pandey and Dushyant Barve (2011). Phytochemical and
Pharmacological review on Annona squamosa Linn. International
Journal of Research in Pharmaceutical and Biomedical Sciences, 2(4):
1404- 1412.
6. Soni H, Malik SS and Singhai AK (2012). Evaluation of leaf of aqueous
extract of MeOH extract of Aannon squamosa on cell viability
American. J. Pharm. Tech. Res., 2(4): 936-944.
7. Abhijit Dey. .Alstonia scholaris R.Br. (Apocynaceae): Phytochemistry
and pharmacology: A concise review.Journal of Applied
Pharmaceutical Science. 2011;01(06): 51-57.
8. Arulmozhi S, Papiya Mitra Mazumder, Sathiya Narayanan L, Prasad
A Thakurdesai. In vitro antioxidant and free radical scavenging
activity of fractions from Alstonia Scholaris linn. R.Br. International
Journal of PharmTech Research. 2010; 2(1):18-25.
9. Sen S, Sahu NP, Mahato SB. Flavonol glycosides from Calotropis
gigantea. Phytochemistry 1992; 31: 2919-2921.
10. Kintzios SE. Terrestrial plant derived anticancer agents and plants
used in anticancer research. Crit Rev Plant Sci 2006; 25: 79-113.

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