Download Content: Introduction

Content:
Introduction………………………………………………………………………………………2
Method and material…………………………………………………………………………16
Results……………………………………………………………………………………… ……19.
-Cytotoxic effect of Nigella Sativa extract……………………………………20
-Cell Viability analyzer……………………………………………………………….20
-Cell Staining and morphology…………………………………………………21
Discussion……………………………………………………………………………………22
Conclusion …………………………………………………………………………………23.
References…………………………………………………………………………………..24
1
Introduction:
Cancer is considered a modern disease of great concern , and the real
cause of this disease is unknown. Some studies mentioned that
exposure to radiation, chemical substances, some drugs, environmental
factors are among the attributed factors . virus infection was also
accused for cancer induction ..
Generally, cancer or tumor is defined as a group of cells that proliferate
rapidly without any control leading to tumor formation in special organ
(local) or spreading to involve any part of the body (like in Leukocyte
Cancer –leukemia -).
Normally, the cells grow and dividing (proliferate) to form new cells
according to the body’s need. When the cells grow old and unable to
function, apoptosis or programmed cell death is the process helping get
rid of such cells .For still unknown caused , apoptosis was shut down by
gene mutation and stimulated growing cells continue to proliferate
without control, with subsequent cancer development
.
There are two types of tumors:
1)Benign Tumors: are not cancer and they have specific characters.
Firstly, they do not spread beyond its origin. Secondly, they do not come
back after they are removed Thirdly, they are rarely killing so it is not a
life threatening..
.
2)Malignant tumors: as their name imply , they are cancer
that characterized by spreading to nearby tissue and organs. Also,
cancer cells can breakaway from a malignant tumors and enter the
bloodstream or lymphatic system. That is how cancer cells spread from
the original source (primary tumor ) to form new masses ( metastasis)
in other organs
2
Uterine cancer is one of the most common cancer between
female. As usually, the exact causes of uterine cancer is unknown.
Logically, this disease like any cancer is not contagious that is mean it
doesn’t have the ability to transfer from person to other.
Figure1
The uterus is part of female’s
reproductive system (uterus- 2ovariesfallopian tubes- cervix-vagina- vagina
opening).Fig ( 1)
It lies in the pelvis between the bladder
and rectum. The narrow, lower portion of
the uterus is the cervix which act is a
conduct between uterus and vagina.
The broad, middle part of the uterus is the
body (corpus). The dome –shaped top of
the uterus is fundus. The fallopian tubes
exited from either top of the uterus to
ovaries to catch the eggs when it matures.
The wall of the uterus consists from 3 layers. The most inner one (lining)
is Endometrium. The middle is formed of smooth muscular tissue
(myometrium). In women of childbearing age, the lining of the uterus
grows monthly and become thick to prepare the uterus for pregnancy
(www.canceranswers.com) .
If there is no pregnancy the most inner functional layer degenerate ,
flows out of the body mixed with blood through the vagina and causes ,
the period or menstrual cycle .The uterus is prone for development of
Fibroids, Endometriosis and Endometrial hyperplasia.
Fibroids: are common benign tumors. They occur mainly in
women in their forties. Women may have many Fibroids at the same
time Fibroids are considered as benign tumors of the uterus
Usually, fibroids cause no symptoms and need no treatment. But
depending on their size and location, fibroids can cause bleeding,
vaginal discharge, and frequent urination. Women with these
symptoms should see a doctor. If fibroids cause heavy bleeding, or if
they press against nearby organs and cause pain, the doctor may
suggest surgery or other treatment.
3

Endometriosis is another benign condition that affects the
uterus. It is most common in women in their thirties and forties,
especially in women who have never been pregnant. It occurs
when endometrial tissue begins to grow on the outside of the
uterus and on nearby organs. This condition may cause painful
menstrual periods, abnormal vaginal bleeding, and sometimes
loss of fertility (ability to get pregnant), but it does not cause
cancer. Women with endometriosis may be treated with
hormones or surgery.

Endometrial hyperplasia is an increase in the number of cells in
the lining of the uterus. It is not cancer. Sometimes it develops
into cancer. Heavy menstrual periods, bleeding between periods,
and bleeding after menopause are common symptoms of
hyperplasia. It is most common after age 40.
To prevent endometrial hyperplasia from developing into cancer,
the doctor may recommend surgery to remove the uterus
(hysterectomy) or treatment with hormones (progesterone) and
regular followup.
Uterine cancer (malignant cancer) are dividing to three types:
1-Endometrial cancer
2-cervical cancer.
3- Uterine sarcoma.
Endometrial cancer is the most common type of uterine cancer. It is a
malignancy which is occur in the endometrium(lining of the uterus).
There is many risk factors that raise the chance to get such like cancer
, excessive exposure to estrogen(hormone replacement therapy), period
of menopause, diabetes and many other factors.
This cancer may be developed from Endometrial hyperplasia. Usallay,
treatment of this disease is radiation therapy, chemotherapy; abdominal
hysterectomy is recommended and hormonal therapy (progesterone
and anti-estrogen).
4
In the present project, we will focus on cancer cervix.
Cervical Cancer is a malignant tumor of female
genital tract (ref)
Risk factors that increases the chance to
get this disease include:
HPV (Human Papilloma Virus) which was
reported to be responsible for the most cases
of cervix cancer, smoking, HIV infection,
Chlamydia infection, multiple pregnancies,
hormonal contrception, exposure to hormonal
drug diethylstilbestrol (DES), and family history of cervical cancer.
The early stage of cervical cancer may be asymptomatic at all.
Symptoms
of cervical cancer are vaginal bleeding, pain during intercours
and vaginal dischare. In the advanced cases, metasis (spread of cancer
cell) may be find in the abdomenand lungs. Symptoms of metasis are:
weight lossm appitie loss,legg pain, single swolleng leg, heavy bleeding
from vagina, leaking of urine and stool from vagina, and bone fractures.
Dignosis
Dignosis of Cervical Cancer depending on
the
Pap smear (Papanicolaou test ):
The pap test is a screening test for
Cervical Cancer . if the result of the pap
smear test is positive, it must apply
confirmatory test Loop Electrical Excision
procedure(LEEP) and coniziton ( cone
biopsy), in that the inner lining of the cervix is removed to be examined
pathologically.
5
Treatment
of Cervical Cancer was designed according to the stage and disease and
severity.
-Early stage (size of tumor less than 4cm) can be treated with radical
hysterectomy with removal of lymph nodes or raition therapy.
- The large early stage (size of tumor more than 4cm) may be treated
with radiation therapy and chemotherapy(cis-platin), hysterectomy with
radiation therapy or chemo(cis-platin) followed by hysterectomy .
- The late stage where tumors are treated with radiation therapy and
chemotherapy(cis-platin).
- Microinvasive cancer is usallay treated with hysterectomy.
Today, the world use a combination of two chemotherapy drugs
(Hycamin+ cisplatin) for women with late stage (IV B) Cervical Cancer
treatment.
Combination Treatment has significant risk of neutropenia (decrease of
neutrophill), Anemia and thrombocytopenia (decrease of platelets).
6
That is right, the chemotherapy and the radiation therapy have benefits
to fight the cancer cells but they also damage normal cells. That is mean
they have risks and disadvantage more than benefits!.
Because all of this risks and harmful side effects, all of the world today
directs to treat many disease especially cancer by herbs. And there are
many researches today that find treat the malignant tumors with
different types of herbs.
The herbs are naturally products (created by Allah) without any human
made. Well known herbs are safe for the body and usually have no side
effects or risks. Herbs are slow in action to get their effect, but often
they are effective. This rule don’t apply on the all of herbs but some
herbs play an important rule in fighting cancer cell or protecting the
body from tumor formation ( by increase the immunity).
In this project, we will focus on an eastern herb that was the most
common herb in medicine in the 14 centuries ago in east. Specially in the
Islamic society; because prophet Mohammed (peace be upon him) said:
“ Black Seeds is a Cure For Every Disease Except Death.” .
Scientific Name of Herb: Nigella Sativa
Other common name: black seeds, black
cumin, the blessing seeds, fennel flower,
black caraway, nutmeg flower, Roman
coriander, or black onion seed and black
sesame.
Organoleptic Characters:
Nigella sativa or blackseed, a member of
the buttercup family (the Ranunculaceae
family).
Nigella sativa is an annual flowering plant.Original home of Nigella
sativa is Asia and Mediterranean region, and it is culturing in
Syria,Iraq,India, Pakistan,Iran, Saudia Arabia and Egypt in Africa.
7
Nigella Sativa seeds
The flowers are coloured pale blue and white,
with 5-10 petals.
The fruit is a large and inflated capsule composed
of 3-7 united follicles, each follicles containing
numerous seeds. The seed is used as a spice.
Nigella sativa has been used for centuries, both
as a herb and pressed into oil, by people in Asia,
Middle East, and Africa for medicinal purposes.
It has been traditionally used for a variety of
conditions and treatments related to respiratory health, stomach and
intestinal health, kidney and liver function, circulatory and immune
system support, and for general overall well-being.
In Islam, it is regarded as one of the greatest forms of healing medicine
available.
The market samples of the seeds of Nigella sativa are
often adulterated. seeds of Argemone mexicana are
often mixed with it.
Nigella sativa seeds are also commonly confused
with onion (Allium cepa L) seeds.
To solve the problems of identification, adulteration
with the help of morphology and micro-morphology
depending upon the nature of the problem, so that
the people may benefit from the real plant.
onion (Allium cepa L)
The seed coat of N. sativa shows pebble-like pattern
with concave surfaces and the seed is black in the shape of a prism or
triangular its umbilical end being smaller than the other, black
In the case of Allium cepa the ornamentation is reticulate.
onion seeds are also black but irregularly shaped.
the seeds of N. sativa are , the umbilical end being smaller than the
other.
The seeds of N. sativa Its odour is strong and taste is bitter
onion seeds is odourless .
8
There are three types of Nigella sativa:
1-the most common type called: Nigella Sativa.
2-Nigella damascene.
3-Nigella orientalis.
Chemical structure:
1-
Essential.oil
i(avr.
0.5%,
thymoquinone
max.
was
component
identified
(up
1.5%)i
as
the
main
to
50%)i
i
p-cymene
(40%)i
i
α-pinene
(up
to
15%)i
i
dithymoquinone
and
thymohydroquinone
Other terpene derivatives were found only in
trace
amounts
Carvacrol,
carvone,
limonene,
4-terpineol,
citronellol
significant (10%) amounts of fatty acid ethyl esters
Fatty
2unsaturated
mainly
oil
fatty
linoleic
acid
oleic
(50
acid
eicodadienoic
–
60%)i
(20%)i
acid
9
acids
(3%)i
dihomolinoleic acid (10%) which is characteristic
for
the
Saturated
fatty
genus
acids
i(palmitic, stearic acid) amount to about 30% or less
i3- Numerous esters
of structurally unusual unsaturated fatty acids with terpene
alcohols (7%)i
i4- Traces of alkaloids
which belong to two different types
i1-isoqinoline alkaloids are represented by nigellimin and
nigellimin-N-oxide
i2-pyrazol alkaloids include nigellidin and nigellicin
i5-Saponins
melantin and alpha-hederin
i6-Protein ,crude fiber, calcium, iron, sodium, and
potassium
palmitic, glutamic, ascorbic and stearic acids; arginine;
methionine; lysine; glycine; leucine; and phytosterols
Generally, Nutritional composition of the seeds breaks
down to 21% protein, 35% carbohydrate, and 36% fat
The two principal bioactive constituents of Nigella sativa
are alpha-hederin and thymoquinone.
In the past few decades Nigella sativa has been under scrutiny for its
anti-bacterial, anti-inflammatory and anti-cancer potential.
Here follows a selection of the many studies examining the effects of
Nigella sativa and its various extracts on cancer.
10
1-Immune System Strengthening
Studies begun just over a decade ago suggest that if used on an ongoing
basis, black seed can play an important role to enhance human immunity,
particularly in immunocompromise patients.
In 1986, Drs. El-Kadi and Kandil conducted a study with human
volunteers to test the efficiency of black seed as a natural immune
enhancer. The first group of volunteers received black seed capsules (1
gram twice daily) for four weeks and the second group were given a
placebo. A complete lymphocyte count carried out in all volunteers
before and four weeks after administration of black seed and the placebo
revealed that the majority of subjects who took black seed displayed a
72% increase in helper to suppresser T-cells ratio, as well as an increase
in natural killer cell functional activity. The control group who received
the placebo experienced a net decline in ratio of 7%. They reported,
"These findings may be of great practical significance since a natural
immune enhancer like the black seed could play an important role in the
treatment of cancer, AIDS, and other disease conditions associated with
immune deficiency states."
These results were confirmed by a study published in the Saudi
Pharmaceutical Journal in 1993 by Dr. Basil Ali and his colleagues from
the College of Medicine at Kin Faisal University.
In the field of AIDS research specifically, tests carried out by Dr. Haq on
human volunteers at the Department of Biological and Medical Research
Center in Riyadh, Saudi Arabia (1997) showed that black seed enhanced
the ratio between helper T-cells and suppresser T-cells by 55% with a
30% average enhancement of the natural killer (NK) cell activity.
2. Anti-histamine activity
Histamine is a substance released by bodily tissues, sometimes creating
allergic reactions and is associated with conditions such as bronchial
asthma.
In 1960, scientists Badr-El-Din and Mahfouz found that dimer
dithymoquinone isolated from black seed's volatile oil, under the name of
"Nigellone," and given by mouth to some patients suffering from
11
bronchial asthma, suppressed the symptoms of the condition in the
majority of patients.
Following the results of this early study, crystalline nigellone was
administered to children and adults in the treatment of bronchial asthma
with effective results and no sign of toxicity. It was observed, however,
that although effective, crystalline nigellone displayed a delayed reaction.
In 1993, Nirmal Chakravarty, M.D., conducted a study to see if this delay
could be attributed to the possibility of crystalline nigellone being an
inhibitory agent on histamine. His hypothesis proved correct. Dr.
Chakravarty's study found that the actual mechanism behind the
suppressive effect of crystalline nigellone on histamine is that crystalline
nigellone inhibits protein kinase C, a substance known to trigger the
release of histamine. In addition, his study showed that crystalline
nigellone decreased the uptake of calcium in mast cells, which also
inhibits histamine release.
The importance of these results are that people who suffer from bronchial
asthma and other allergic diseases may benefit from taking crystalline
nigellone.
3. Anti-tumor principles
A study of black seed's potential anti-tumor principles by the Amala
Research Center in Amala Nagar, Kerala (India) in 1991 lent further
impetus to Dr. Chakravarty's suggestion for the possible use of black seed
in the treatment of cancer.
Using an active principle of fatty acids derived from black seed, studies
with Swiss albino mice showed that this active principle could completely
inhibit the development of a common type of cancer cells called Ehrlich
ascites carcinoma (EAC). A second common type of cancer cells,
Dalton's lymphoma ascites (DLA) cells were also used.
Mice which had received the EAC cells and black seed remained normal
without any tumor formation, illustrating that the active principle was
100% effective in preventing EAC tumor development.
Results in mice who received DLA cells and black seed showed that the
active principle had inhibited tumor development by 50% less compared
to mice not given the active principle.
12
The study concluded, "It is evident that the active principle isolated from
nigella sativa seeds is a potent anti-tumor agent, and the constituent long
chain fatty acid may be the main active component."
4. Anti-bacterial
In 1989, a report appeared in the Pakistan Journal of Pharmacy about
anti-fungal properties of the volatile oil of black seed. 1992 saw
researchers at the Department of Pharmacy, University of Dhaka,
Bangladesh, conducting a study in which the antibacterial activity of the
volatile oil of black seed was compared with five antibiotics: ampicillin,
tetracycline, cotrimoxazole, gentamicin, and nalidixic acid.
The oil proved to be more effective against many strains of bacteria,
including those known to be highly resistant to drugs: V. cholera, E. coli
(a common infectious agent found in undercooked meats), and all strains
of Shigella spp., except Shigella dysentriae. Most strains of Shigella have
been shown to rapidly become resistant to commonly used antibiotics and
chemotheraputic agents.
In light of the above research findings, it is of interest that homeopaths
have long been known to make a tincture from the black seed for
digestive and bowel complaints. Traditionally, the black seed is still used
to help relieve vomiting and diarrhea, as well as flatulent colic, and to
help counteract the griping action of purgatives (e.g. certain laxatives,
fruits such as apricots when over consumed).
5. Anti-inflammatory
As early as 1960, Professor El-Dakhakny reported that black seed oil has
an anti-inflammatory effect and that it could be useful for relieving the
effects of arthritis.
In 1995, a group of scientists at the Pharmacology Research Laboratories,
Department of Pharmacy, Kings College, Lond, decided to test the
effectiveness of the fixed oil of Nigella sativa and its derivative,
thymoquinine, as an anti-inflammatory agent. Their study found that the
oil inhibited eicosanoid generation and demonstrated anti-oxidant activity
in cells.
The inhibition of eicasanoid generation, however, was higher than could
be expected from thymoquinone alone. Their study suggested that other
13
compounds within the oil might also be responsible for the enhanced antiinflammatory reactions in cells.
The scientists speculated that the unusual C20:2 unsaturated fatty acids
contained in black seed were possibly responsible for boosting the oil's
effectiveness.
In 1997, studies conducted at the Microbiological Unit of the Research
Center, College of Pharmacy, King Saud University, Riyadh, Saudi
Arabia, found that externally in an ointment form, the anti-inflammatory
activity of the black seed was found to be in the same range as that of
other similar commercial products. The tests also demonstrated that the
black seed is non-allergenic.
6. Promotes lactation
A study by Agarwhal (1979) showed that black seed oil increases the
milk output of breastfeeding mothers.
A literature search by the University of Potchefstroom (1989), including
biological abstracts, revealed that black seed's capacity to increase the
milk flow of nursing mothers could be attributed to a combination of lipid
portion and hormonal structures found in the black seed.
In vitro studies performed in Jordan and the United States
have determined that the volatile oil is anti-leukemic.
Studies performed in Spain as well as England found that
the fixed oil is useful in the treatment of rheumatism and
other inflammatory diseases. This property is attributed to
thymoquinone which is as high as 25% in the Egyptian
seed and missing entirely is some seeds.
14
* A June 2007 study (abstract: http://tinyurl.com/2zksw6),informs us that
“Nigella sativa has immunopotentiation and antihistaminic, antidiabetic,
anti-hypertensive, anti-inflammatory, and antimicrobial activities. [...]
Furthermore, blackseed preparations may have a cancer chemopreventive
potential and may reduce the toxicity of standard antineoplastic drugs.”
The study looked at the in vitro and in vivo (mice) potential of different extracts of
Nigella sativa seeds against several tumour cell lines. The essential oil injected
directly into solid tumours inhibited their development, and even decreased their
volume after 30 days of treatment. The study concludes: “Our results indicate for the
first time that intra-tumor treatment of tumor-bearing mice with essential oil may have
led to the inhibition of metastasis development [...]. These results demonstrate either
that the essential oil has an anti-metastatic activity in mice or that it inhibits or delays
metastasis by rapid reduction of primary tumor volume at the site of induction. [...]
The present study demonstrates that the cytotoxic activity of blackseed extracts is a
complex phenomenon depending not only on the nature of the extract and its
components, but also on the tumor cell type.”
* An August 2007 study published in “Cancer Research”
(http://tinyurl.com/266rlg) examines the in vitro and in vivo effects of one of
Nigella sativa’s active compounds, thymoquinone, on prostate cancer,
concluding that it may prove to be effective in treating hormone-sensitive as well as
hormone-refractory prostate cancer. Furthermore, because of its selective effect on
cancer cells, we believe that thymoquinone can also be used safely to help prevent the
development of prostate cancer. The cytotoxicity of Nigella sativa purified extracts,
thymoquinone (TQ) and dithymoquinone (DIM), against a variety of tumour cells had
already been examined in 1998 (http://tinyurl.com/33woyf).
* A 2005 article (http://tinyurl.com/2u9arx) mentions “the protective effects of
TQ and the volatile oil against the nephrotoxicity and hepatotoxicity
induced by either disease or chemicals. The seeds/oil have antiinflammatory,
analgesic, antipyretic, antimicrobial and antineoplastic activity. The oil decreases
blood pressure and increases respiration. Treatment of rats with the seed extract for up
to 12 weeks has been reported to induce changes in the haemogram that include an
increase in both the packed cell volume (PCV) and haemoglobin (Hb), and a
decrease in plasma concentrations of cholesterol, triglycerides and glucose.”
* Back to Science Daily. Researchers at Thomas Jefferson University
recently discovered that an extract of Nigella sativa, called
thymoquinone, blocked pancreatic cancer cell growth and
killed the cells via apoptosis.
15
Method and Material:
Materials:
-
Nigella Sativa Extract (essential oil, Volatile oil-thymoqunine-).
Hela cells (cancer cervix cells)from tissue culture unit .KFMRC .
Cisplatin prepared by KAUH pharmacy.
Dulbecco's modified eagle Medium (DMEM) was purchased from
MP Biomedicals Inc, USA.
- Fetal calf serum (FCS) was obtained from Gibco.
- Dimethyl sulfoxide (DMSO) and sulforhodamine B were obtained
from (MP Biomedicals, Illkrich, France).
Methods:
Nigella Sativa Extract:Nigella sativa seeds obtained from local market (spice dealer and mills
shop) in Jeddah city.
The Nigella Sativa volatile oil extract prepared in the Lab of
pharmaceutical preparation, collage of pharmacy KAU Jeddah.
The powdered seeds were extracted with MeOH (5 X 1000
ml) by using Ultraturrax T25 homogeniser (Janke and Kunkel, IKA
Labortechnik, Stauten, Germany) at room temperature. The combined
extracts were evaporated under reduced pressure and, lyophilized and
protected from light at 4 oC until use. The perecentage yields for ......
.and types were .... % and ........, respectively.
after that Dissolved in DEMSO (Dimethyl sulfoxide) immediate before
used.
16
Cell Culture:
Hela cell line (derived from cervical cancer cells) was used in this study, and
available in the cell bank of the virology unit, KFMRC, King Abdulaziz
(KAU), Jeddah.
Cell line was grown in DMEM media supplemented with 10% FCS,
penicillin (100 u/ml) and streptomycin (100 ug/ml) in culture flasks at 37 0C
in 5% humidified CO2 incubator. Cells were fed until confluence.
Cytotoxic activity of nigella sativa on the proliferation of Hela
cancer cell line by sulforhodamine (SRB) B assay:
The cytotoxicity of the Nigella Sativa was tested using the method of
Skehan et al., (1990).
Cells were cultured in 96 plate (104 cells/well) for 24 hr in a humidified
CO2 incubator at 37ºC.
Different concentrations of the Nigella Sativa and Cisplatin
(2.5, 7.5, 10, 50 and 100 µg/ml)
were added to the cells and reincubated for 48 hr at 37ºC in a
humidified 5% CO2 incubator. Cells were fixed with 50 µl of cold 50%
trichloroacetic acid (TCA) on the top of growth media in each well. The
cultures were incubated at 4ºC for 1 hr and then washed five times with
tap water to remove TCA. TCA-fixed cells were stained for 30 minutes
with 0.4% (wt/vol) sulforhodamine B (SRB) dissolved in 1% acetic acid. At
the end of the staining period, SRB was removed and cultures were
quickly rinsed four times with 1% acetic acid to remove unbound dye.
Bound dye was solubilized with 10 mM unbuffered Tris EDTA
(100 µl /well) for 5 minutes on a gyratory shaker.
17
Color intensity was measured immediately in an ELISA reader at wave
length 490 – 530.
Cell Viability assay:
The viability of the cells was assessed by cell viability analyzer
which provides an automatic and effective ways to perform the trypan
blue dye exclusion method. Where trypan blue selectively stains dead
cells in which the plasma membranes has lost their integrity, where
the living cells with intact membranes cannot absorb this dye.
cells suspended in original DMEM containing 10% FBS (1×105 cell/ml)
were cultivated onto 24-well plate, each well contain 1ml. After 24h the
original media was replaced by fresh (Fetal Calf Serum) FCS that contain
several concentrations of Nigella Sativa Seeds extracts and
Cisplatin(1:10, 1:100, 1:1000) The controls extract contain 0.1% DMSO.
18
Results:
Cytotoxic activity of nigella sativa on the proliferation of Hela
cancer cell line was shown in Fig ( 5) :
Figure5
Fig. (5) showed the intial significant decrease in hela cells
survival rate at low concentration of NS extract. More higher
concentration were still effective at higher concentration in case of NS
(Qassim) treated cells .
by measure the number of adhesion protein:
Table1
Table (1) . showing that NS ( Qassium ) decrease the adhesion protein of Hela cells in a dose dependant
manner. Similar results were obtained by NS from Ethiopia region.
19
Cell Viability analyzer:
Nigella Sativa Nigella Sativa
Cisplatin (Ethiopia)
(Qassim)
Control concentration
0.077
0.173
0.167
0.21
1:10
0.22
0.027
0.086
0.21
1:100
0.1
0.303
0.177
0.21
1:1000
20
Cell morphology :
Using Comassi Stain ,the morphology of cancer Hela cells proved
the anti cancer effect of NS extract .
Shrinkage and apoptotic changes were observed and it was more
marked with Ns Qassim
Hela cells after 24 hrs without
any treatment
Hela cells after incubated for 24 hrs
with 1/100 concentration of NS Ethiopia
Hela cells after incubated for 24 hrs
with 1/100 concentration of cisplatin
Hela cells after incubated for 24 hrs
with 1/100 concentration of NS Qassim
21
Discussion:
Although the quinone TQ has demonstrated significant in vitro
and in vivo antineoplastic activities against different tumor cell
lines, the mechanism of this effect has not been fully examined
(8,19).
This compound's ability to effectively-kill several types of tumors
without significant cytotoxicity to normal cells(10)
In this study, TQ demonstrated selective cytotoxicity
in vitro for human cervical cncer cells tested in comparsion with
Cisplatin. Moreover, TQ showed the greatest toxicity towards
Hela cells more than Cisplatin toxicity.
The morphological results of this study suggest that TQ inhibited
proliferation of tumor cells by a mechanism that involves cytotoxicity and
apoptosis. Significant of Cell Viability was seen only after
treatment with 1/100
We studied the early cellular responses induced by TQ
treatment in an attempt to define the events associated with
cytotoxicity.
Significantly, we showed by Comassi stain in hela cells that has been
treated with NS Ethiopia a markedly decrease in cell density per field
and shrinkage cells more than seen in NS Qassim and Cisplatin.
The most likely reason for this observation may be NS Ethiopia contain
more TQ than Qassim.
22
Conclusion:
These results suggest that TQ may be a promising new anticancer
therapeutic agent and a potential candidate to be further evaluated.

Thymoquinone is the active form of Nigella Sativa extract, it is
effective in fighting cervical cancer cells (hela cells).

Nigella Sativa Extract is more effective in comparsion with
Cisplatin.

Nigella Sativa of Qassim is more effective than NS Ethiopia.
23
References:
1- Nanda, Rita (2006-06-09). "Cervical cancer". MedlinePlus Medical Encyclopedia.
National Institutes of Health.
http://www.nlm.nih.gov/medlineplus/ency/article/000893.htm. Retrieved 2007 -1202.
2- On June 15, 2006, the US Food and Drug Administration
3-www.wikipidia.com
4- http://www.healthymuslim.com/articles/mdhpn-black-seed-oil-nigella-sativa-andits-disease-preventing-effects.cfm
5- http://margaret.healthblogs.org/other-alternative-treatments/nigella-sativablack-cumin/
6- http://www.kitchendoctor.com/herbs/black_cumin.php
7- http://www.medicinenet.com/uterine_cancer/article.htm
http://www.health.com/health/library/topic/0,,tm6524_tm6525,00.html
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