Download Though our project did produce some positive results, there were

Analyzing The Effect Of Perillyl Alcohol And Pure Organic Lavender
On Breast Cancer Cells
Rock, Esther and Vlasic, Ana
Suffolk County Community College
ABSTRACT
RESULTS
Perillyl alcohol is a monoterpene isolated from the essential oil of lavender. Perillyl extract
has been shown to exhibit chemotherapeutic, and chemo preventive potential in breast cancer cells.
Perillyl alcohol, and lavender essential oil were chosen for comparison of toxicity on MDA-MB231, Estrogen receptor negative, and MCF-7, Estrogen receptor positive cancer breast cells grown in
culture.
Apoptosis was induced in these cell lines with perillyl alcohol, and pure Lavender extract at
micro molar concentrations. Our results show that perillyl alcohol alone is more potent than lavender
extract. This compound shows potential as a cancer therapy to fight a variety of breast cancer in vivo.
From our experiment, we were able to conclude that because the pure lavender was at a higher
concentration, the results of apoptosis were seen in earlier stages. In using the proliferation assay, which
allowed us to determine the number of cells that are growing in the absence or presence of certain
proliferation affecting agents, we were able to determine that for the POH, the concentrations of
3.75um, 7.5 um, and 10 um, yielded the less amount of cells. In using the pure Lavender, we received
results of 10 um, and 25 um causing most apoptosis.
All together, better results were seen in the MCF-7 breast cell lines. Those cells treated with the
pure lavender presented quicker results, though the POH was more potent.
We also tested our cells by doing an adhesion assay, which shows cell binding when a drug
is, or isn’t added. Though not excellent, we did receive some promising results of adhesion decreasing
in cancer cells treated with the lavender drug.
DISCUSSION
Though our project did produce some positive results, there were many things that if had
been done correctly, would have enhanced our results. We intended to do three separate
assays; proliferation, adhesion, and migration, but the migration, and adhesion assays did not
produce any results. Mistakes from incorrect media, and calculation inaccuracies, left our
experiment with no cells to continue with. We based our results solely on our proliferation
procedure, and the effects of the drugs on the cells from the adhesion assay.
MCF-7 PO H 7, 10 um
CONCLUSION
INTRODUCTION
Cancer is the second leading cause of death in the United States. Cancer strikes one in every
two men and more than one in every three women, .
Many of the cancer drugs used are effective in treating the beginning stages of cancer, but none
can cure metastatic disease. These drugs also cause adverse side effects, which may include nausea,
vomiting, loss of hair, loss of appetite, and a suppressed immune system. It is for this reason that
alternative therapies are being considered. Alternative therapies such as, Perillyl Alcohol, extracted
from Lavender, are being tested for the possible application in inducement of apoptosis of cancerous
cells, and these natural treatments would also have no adverse side effects.
This experiment will compare the effect of pure organic Lavender and Perillyl extract, a
lavender extract, to see if the pure organic lavender will have the same or comparable effect, if any, in
inducing apoptosis on cancerous cells.
MCF-7 POH 1, 1.25 um
METHODS
Proliferation Assay :
(28 x 10 4Cells were tryptonized and seeded in 96 well plates ). Cells were then incubated for 48 hours.
MTT assay is based on cleavage of the yellow tetrazolium salt MTT to purple formazan crystals by
Metabolic active cells. The cellular reduction uses the pyrimidine nucleotide cofactors NADH and
NADPH. The crystals formed result in a colored solution , and is quantified using a scanning multi wells
Spectrophotometer (ELISA reader). This is used for accuracy, online computer processing of data, which
allows for a rapid handling of high number of samples. (ROCHE) The cellular reduction uses the
pyrimidine nucleotide cofactors NADH and NADPH. The crystals formed result in a colored solution and
is quantified using a scanning multi well spectrophotometer (ELISA reader).
This is used for accuracy, online computer processing of data, which allows for a rapid handling of high n
umber of samples. (ROCHE)
Adhesion assay:
ell adhesion assays were performed with fibronectin coated 96-well cluster plates.
Wells were washed with PBS with 0.2% Tween-20, blocked with 5% blotto (PBST with 5%
skim milk). Then washed twice with PBST. Cells were then seeded at a concentration of .58x 10^6
with and without migration media, perillyl alcohol, and lavender essential oil. Both cell lines were
incubated at 37 degrees Celsius for 30 minutes. The MCF-07 with CO2. Unbound cells were dislodged
by inverting, submerging, and rocking the plate in PBS for 15 minutes. Cells were then fixed in
formaldehyde, stained with crystal violet, and lysed with 1% SDS.
MCF-7 POH control
MCF-7 lavender 1 1um
MDA-MB 231 lavender control
MDA-MB 231 Lavender 10 um
Special Acknowledgement:
Williams Jennie PhD, Moloney Daniel PhD, Daccueil Farah MS, Grau Fred MAT
Funding provided by , NIH GM 50070
MDA-MB 231 Lavender 25um
MDA-MB231 , 2, 2.5 um
MCF-7 lavender 3, 10 um
MDA-MB231, POH 4, 3.75 um
Our focus in deciding to analyze, and compare Perillyl Alcohol against Pure Organic Lavender
essential oil was to see if Lavender would not only have an effect against cancer cells, but
work better than the Perillyl Alcohol. MDA-MB 231, and MCF-07 breast cancer cells lines
were used.
We learned that both the Perillyll Alcohol, and the pure Lavender Essential Oil worked on both
cancer lines. The best results seemed to be on the MDA-MB 231 line. The Lavender oil
worked with higher concentrations on both cells lines, while the Perillyl Alcohol worked in
smaller doses.
During this experiment, we applied cell proliferation, and adhesion assays. The proliferation
assay worked, and results were positive for both the Perillyl and the Lavender. The adhesion
assay was repeated three times. The first time, we had an error in the concentrations used. The
second and third failed due to the type of media used.
In conclusion, changes on the cancerous cells were seen with both the Perillyl Alcohol, and the
Pure Lavender Essential Oil. Even though our results were not perfect, and we were limited on
time, the experiment provides a positive base for further analysis of these alternative
therapies.
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