Download Programming Cancer Cell Death

+
Professor Davis
Counselors:
Alex McGovern
Taylor Szupiany
BASE Camp
2012
Campers:
Jobeenscya
Alexis
Jennifer
Castro
Gabrielle
Edwards
Gabriela
Rodriguez
Shennell
Tyndell
+
Introduction to Cancer
n 
Non-Cancerous cells divide when needed; however,
cancerous cells divide continuously and results in a tumor.
n 
The constant division is due to a mutation in a cell
n 
n 
Mutated genes are called oncogenes
The mutation of oncogenes can impair the function of healthy
cells.
+
Signal Transduction
Signal Transduction is a process that the cell must
undergo in order for the cancer cell to successfully form.
Signal to
grow is
turned on
Signal to
stop
growing is
turned off
Signal to
die is
turned off
Understanding Signal Transduction helps us understand
which step to eliminate in order to induce apoptosis.
Cancer
+
Introduction to Cancer: Leukemia
n 
In this experiment, we treated Leukemia cells with our
products
n 
Leukemia is a type of cancer that is within the blood cells.
n 
Characterized by uncontrolled division of white blood cells which
results in abnormalities or death of red blood cells
Because Red blood cells
take oxygen to the body, it
may lead to anemia,
abnormal bleeding and
infections.
.
+
Apoptosis:
§ 
Apoptosis is when cells tell themselves to commit
suicide due to a problem or age. Also known as
programed cell death.
+
Purpose, Hypothesis,
Prediction
n  Purpose- To find natural treatments for killing
leukemia cancer cells
n  Hypothesis- If natural products are applied to cancer
cells apoptosis will be induced
n  Prediction- If apoptosis is induced in cancer cells then
the fraction of cytochrome c in the cytosol over the
mitochondria will be greater then in the control cells
+
Procedure:
Treat
Collect
ELISA
n 
Centrifuged the supernatant to a high g force to separate the
mitochondria.
n 
Collected the supernatant as the cytosol fraction and collect
the pallet as the mitochondria.
n 
An ELISA was done to specify the amount of Cytochrome C in
the cytosol.
+
Antibody
Cytochrome C
Stop Soln
Cyt C Antibody
with Biotin Wash
Wash and
Substrate
ENZYME LINKED IMMUNABSORBANT ASSAY
+
n 
Molecules of natural origin
n 
Evolved for defense
n 
Make up the majority of drugs today:
-78% of antibiotics
-74% anticancer
n 
Complex Structures; lack of abundance
+
Cinnamon has extremely high
anti-oxidant activity, and the
oil of cinnamon has strong
anti-bacterial and anti-fungal
properties.
Ginger root is effective in
reducing nausea and also may
be useful in reducing the pain,
stiffness, and immobility of
arthritis
+
Milk thistle is a potent
antioxidant, it significantly
increases levels of glutathione,
which the liver uses to detoxify
and metabolize harmful
substances.
Red clover has been used to
treat coughs and respiratory
system congestion, because it
also contains resin. Red
clover's constituents are
thought to stimulate the
immune system.
+
Garlic displays antibiotic,
antifungal, and antiviral
properties and is reportedly
effective against many flu
viruses and herpes simple.
Aloe’s antiseptic and
antibiotic properties allow its
use even in more severe skin
problems, such as second
degree burns, eczema and
severe sunburn.
+
Melatonin is secreted by the
pineal gland in the brain, and
it has been used to treat a
number of sleep-cycle
problems, and other medical
illnesses.
Green tea contains very
powerful antioxidants. These
destroy the free radicals in
your blood stream, preventing
them from prematurely aging
your skin and damaging your
eyes.
+
n 
SNAP is a stable green crystalline compound that mimics the
action of nitric oxide the chemical that triggers cell death
+
Results
Cyt to Mit Ratios
Cyt to Mit Ratios
25
15
Series
10
5
0
Natural Product Treatment
Cyt/ Mit
Cyt/ Mit
20
90
80
70
60
50
40
30
20
10
0
Cyt/Mit
Percent
Dead
Natural Product
+
Discussion
n 
Most effective treatment: Green Tea
n 
15% error
Greatest proportion of Cytochrome C in the Cytosol (19)
n 
Future experimentation should involve green tea
n 
n 
n 
Least effective treatment: Garlic
n 
7.3% error
n 
Smallest proportion of Cytochrome C in the Cytosol (.109).
Most precise treatment: Milk Thistle
n 
n 
0% error
Overall, not an effective treatment.
+
Discussion continued: Future
Experimentation
n 
Green tea proved to be the most effective treatment
n 
Future experimentation
n 
More accurate/precise results should be obtained to ensure
reproducibility
n 
Experiments involving green tea.
n  Determination/extraction of active ingredient(s).
n 
Specific anti-cancer experimentation
n 
Different cells
+
Discussion: Error
n 
Human Error:
n 
n 
n 
Systematic Error:
n 
n 
A mistake made by the experimenter
Should not appear in a lab report; if
mistakes are made, experiment should be
repeated
Mistake/s in the procedure
Random Error:
n 
Imperfect measurements, result of limited
measuring equipment
+
Sources of Error
n 
Human Error:
n 
n 
n 
n 
Potential misuse of pipetter
When incubated, samples given different beginning times, but
received same end time.
Systematic Error:
n 
Cytochrome C was outside of range of line (1.3), needed to be
diluted (3 dilutions).
n 
Different number of cells per test
Random Error:
n 
Pipetter
+
Acknowledgements
n 
NIH Grant
n 
Fairfield University Chemistry and Biology Department
n 
Dr. Davis
n 
Dr. Phelan
n 
Alex and Taylor !