Essential Oils and Cancer Research 1

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Essential Oils andCancer Research (1)
YLEO 2007 Convention Sue Chao Research Lab
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Top Five Causes of Death in U.S.1

• Number of deaths for leading causes of mortality
• Heart disease 654,092
• Cancer 550,270
• Stroke (cerebrovascular diseases) 150,147
• Chronic lower respiratory diseases 123,884
• Accidents (unintentional injuries) 108,694

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Top Five Most Commonly Diagnosed Cancers in U.S.2
Male
Female

• Total Cases 2,671,629
• Breast 30.4
• Lung 13.3
• Colon Rectal 11.7
• Corpus Uterus 5.7
• Non-Hodgkin
• Lymphoma 3.9

• Total Cases 2,853,795
• Prostate 29.4
• Lung 15.8
• Colon Rectal11.2
• Bladder 6.5
• Non-Hodgkin
• Lymphoma 4.1

All cancers are caused by uncontrolled
proliferation of cells
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Cancer Research Common Methods of
DeterminingCellular Proliferation

• Radioactive labeling of DNA synthesis 3-4
• SRB sulfurhodamine B assay 5-6
• MTT tetrazolium salt assay 7-8

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The Principle of Radioactive Proliferation Assay
DNA Synthesis
G2
S
M
G1
Interphase
Mitosis
DAUGHTER CELLS FORMED
Figure 1 Cell Cycle
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The Principle of Radioactive Proliferation Assay
9-11

• The division of all normal human cells occurs
  during the cell cycle and is dependent upon DNA
  synthesis (Figure 1).
• There are two phases in the cell cycle
  interphase and mitosis phase
• - Interphase includes three stages
• G1 phase Cell grows
• S phase DNA is synthesized and chromosomes are
  replicated
• G2 phase Cell continues to grow and prepares
  for mitosis
• - Mitosis phase Prophase, prometaphase,
  Metaphase, Anametaphase and Telophase. During
  mitosis, cell division occurs where one
  parent cell becomes two daughter cells.

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The Principle of Radioactive Proliferation Assay

• The more cell divisions, the more radioactivity
  will be incorporated into the DNA and more cells
  will be counted.
• During cell harvesting, the cells DNA is set
  free when cells are lysed by water. Intact DNA,
  millimeters in length, are retained on fiber
  glass filters
• The amount of radioactivity in radio-labeled DNA
  is counted in a scintillation counter.
• Inhibition of proliferation by a growth inhibitor
  is calculated by
• Inhibition of proliferation (cpm
  untreated cpm treated) / cpm untreated ? 100

cpm counts per minute
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Material

• Lung Cancer Cell ATCC CCL 185 12-13
• Normal Cell ATCC CCL 64
• Label marker 3H Thymidine
• Dye Trypan Blue
• DMSO Oil dilution agent
• Essential Oils Testing agent

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Materials - 1
Lung Cancer Cell
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Materials - 2
Normal Lung Cell
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Materials - 3
Trypan Blue
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Materials - 4
3H-Thymidine
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Materials - 5
DMSO
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Materials - 6
Young Living Essential Oils
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Procedure

• Day 1 - Cell culture (Photo 1 - 6)
• Day 2 - Cells are resuspended at a certain
  density (Photo 7 10)
• - YLEOs are diluted to various dilutions
  (Photo 11, 12)
• - Dilutions of YLEOs are added to cells
  (Photo 13)
• Day 3 - Add 3H-thymidine to label DNA synthesis
  (Photo 14)
• Day 4 - Harvest cells (Photo 15)
• - DNA fragments are retained on fiber glass
  filters (Photo16)
• - Drying glass filters (Photo 17,18)
• - Determine radioactivity (cpm) of each
  sample (Photo 1923)

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Procedure Day 1
Photo 1 cell culture
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Procedure Day 1
Photo 2 cell culture
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Procedure Day 1
Photo 3 cell culture
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Procedure Day 1
Photo 4 cell culture
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Procedure Day 1
Photo 5 cell culture
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Procedure Day 1
Photo 6 cell culture
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Procedure Day 2
Photo 7 cell preparation
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Procedure Day 2
Photo 8 cell preparation
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Procedure Day 2
Photo 9 cell preparation
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Procedure Day 2
Photo 10 cell preparation
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Procedure Day 2
Photo 11 oil preparation
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Procedure Day 2
Photo 12 oil preparation
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Procedure Day 2
Photo 13 add essential oil to cells
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Procedure Day 3
Photo 14 radioactive labeling of DNA synthesis
using 3H-thymidine
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Procedure Day 4
Photo 15 harvesting cells
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Procedure Day 4
Photo 16 DNA fragments are retained on fiber
glass filters
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Procedure Day 4
Photo 17 drying fiber glass filters
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Procedure Day 4
Photo 18 drying fiber glass filters
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Procedure Day 4
Photo 19 preparing to count samples
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Procedure Day 4
Photo 20 preparing to count samples
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Procedure Day 4
Photo 21 scintillation analyzer
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Procedure Day 4
Photo 22 counting samples
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Procedure Day 4
Photo 23 cell count
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Results
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Results
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Figure 1 Twelve EOs (with at least one dilution)
with inhibitory effects of greater than 50 on
cancer cells, and less than 20 inhibitory
effects for the normal cell line
inhibition
CCL-185 Lung Cancer CCL64 Normal Cell
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Table 3 The major components of 12 Essential Oils
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Table 4 The inhibition effects of Mt. Savory Oil
on lung cancer cell proliferation is
dose-dependent
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Table 5 All dilutions of Y.ylang and Valerien
oils significantly inhibited the proliferation of
the lung cancer cells but had minimal effects on
the normal cell line
Figure 3 All dilutions of Y.ylang and Valerien
oils significantly inhibited the proliferation of
the lung cancer cells but had minimal effects on
the normal cell line
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Conclusion

• 21 out of 38 essential oils inhibited cancer
  cell proliferation by more than 50
• 26 out of 38 essential oils inhibited normal
  cell proliferation by less than 20
• 12 of 38 essential oils at various dilutions
  inhibited cancer cell proliferation by more than
  50, but only inhibited the proliferation of
  normal cells by less than 20.
• - These oils include Cistus, Clove, Douglas Fir,
  White Fir, Lavender, Nutmeg, Patchouli, Palo
  Santo, Tangerine, Valerian and Ylang Ylang.
• Valerian and Ylang Ylang oils have the broadest
  range of dilutions that exhibit inhibitory
  effects on cancer cell growth
• The inhibitory effect of essential oils on
  cancer cell growth is dose-dependent
• Further research on the optimal dose of essential
  oil with inhibitory effects on cancer cell
  proliferation is needed for clinical purposes

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References

• 1. Deaths Preliminary Data, 2004 National
  Vital Statistics Reports NVSS vol 54 No.19
• 2. Top Five Most Commonly Diagnosed Cancers in
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