introduction chili peppers are eaten throughout the world in a variety of dishes, and cuisines...

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Introduction Chili peppers are eaten throughout the world in a variety of dishes, and cuisines Capsaicin, an active component in chili peppers, is responsible for the burning sensation of peppers Capsaicin in peppers has been used in cancer research and shows that it is able to inhibit the growth of prostate cancer cells The prostate cells were subjected to varying doses of capsaicin for 24 hours and apoptosis was observed in a dose dependent manner (0 µM- 1000µM) (Mori et al. 2006) In a similar study, it was found that capsaicin suppressed the growth of leukemic cells, but not healthy mononuclear bone marrow cells The study stated that the capsaicin did this by stopping the cell in the G 0 -G 1 phase and allowing the cell to go through apoptosis Leukemic cells were exposed to varying levels of capsaicin (0 µM-1000µM) over a 24 hour period in addition to subjecting cells to a dose of 50 µM for 0 to 120 hours These tests suggested that the cells responded to capsaicin in a dose/time dependent manner The higher/longer the dose of capsaicin, the faster the cells went through apoptosis The study also discovered that when capsaicin was injected into tumors, the tumors would shrink compared to control tumors Mice were used as an in vivo test group in both experiments, and were not detrimentally affected by the capsaicin (Ita et al. 2004). Works Cited Ito, Keisuke., Nakazato, Tomonori., Yamato, Kenji, 2004. Induction of Apoptosis in Leukemic Cells by Homovanillic Acid Derivative, Capsaicin, through Oxidative Stress: Implication of Phosphorylation of p53 at Ser-15 Residue by Reactive Oxygen Species. Cancer Research [serial online] 64: 1071-1078. Mori, Akio., Lehmann, Soren., O’Kelly, James, 2006. Capsaicin, a Component of Red Peppers, Inhibits the Growth of Androgen-Independent, p53 Mutant Prostate. Cancer Research [serial online] 66: 3222-3229. Methods Hypothesis H 0 :Treating MCF-7 breast cancer cells with Capsaicin will not cause cell death. H 1 : Treating MCF-7 breast cancer cells with Capsaicin will induce cell death through apoptosis. Chili Peppers: A Cure for Breast Cancer? Treating MCF-7 Breast Cancer Cells with Capsaicin at Varying Concentrations Andrew Sutton Department of Biology, York College of Pennsylvania Conclusion Cultured MCF-7 cells with DMEM + 10% Fetal Bovine Serum + 1% Penstrep + 1% GlutaMAX Allowed cells to reach confluency Removed cells from 30 mL flask using trypsin Counted cells using a hemocytometer, diluted to 10^ 6 cells/mL 150 µL of cells were put into each well of 96 well microtiter plate Cells were allowed a day to attach and multiply After first day, cells were administered doses of Capsaicin varying from 0µM to 1000 µM Capsaicin was dissolved in 100% EtOH or DMSO Equivalent concentrations of solvent were tested Four replicates of each concentration were tested Control healthy cells received 50 µL of growth media Control dead cells received 50 µL of cyclohexamide or NaN 3 Cell death measured using MTS/PMS assay Absorbance was measured, the greater the absorbance the more living cells were present Also ran a Caspase 3 assay to determine if cell death was from apoptosis or necrosis The higher the fluorescence, the more Caspase 3 protein present suggesting the cells went through apoptosis Concentrati on Amt. of Capsaicin /EtOH (uL) Amt. of Media (uL) Total Amt. of Capsaicin/ EtOH (uL) Total Amt. of Media (uL) 0 uM 0 50 0 250 1 uM 0.01 49.99 .05 249.95 10 uM 0.1 49.9 0.5 249.5 50 uM 0.5 49.5 2.5 247.5 100 uM 1 49 5 245 200 uM 2 48 10 240 500 uM 5 45 25 225 750 uM 7.5 42.5 37.5 212.5 1000 uM 10 40 50 200 Resul ts Capsaicin was able to kill the MCF-7 breast cancer cells in dependent manner Control cells were minimally affected by the solvent Caspase 3 Assay suggests apoptosis as mode of cell death Future Studies Does Capsaicin effect all cancers the same way? Does Capsaicin effect bacteria? Acknowledgements Thank you Dr. Thompson for all your guidance on this project. Thank you to Joan Carpenter for obtaining the M tp://en.wikipedia.org/wiki/File:Kapsaicyna.svg 7 cells in culture dishes prior to various Capsaicin treatments for Caspase 3 Assay. -7 cells growing and dividing at 400X magnification (left) and 200X (right). Figure 3: MCF-7 cells after treatment with Capsaicin at varying concentrations. 0 mM 1 mM Increasing concentration Figure 5. Caspase 3 Assay of Varying Levels of Capsaicin Treated MCF-7 cell after 24 hours. 500 μM Capsaicin treatment showed a significant difference to an unpaired t-test. P-value <0.05 ns ns ns * Figure 4. Treatment of MCF-7 Cells with Capsaicin at various mM concentrations

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Page 1: Introduction Chili peppers are eaten throughout the world in a variety of dishes, and cuisines Capsaicin, an active component in chili peppers, is responsible

Introduction• Chili peppers are eaten throughout the world in a variety of dishes,

and cuisines• Capsaicin, an active component in chili peppers, is responsible for the

burning sensation of peppers• Capsaicin in peppers has been used in cancer research and shows

that it is able to inhibit the growth of prostate cancer cells• The prostate cells were subjected to varying doses of capsaicin for 24

hours and apoptosis was observed in a dose dependent manner (0 µM-1000µM) (Mori et al. 2006)

• In a similar study, it was found that capsaicin suppressed the growth of leukemic cells, but not healthy mononuclear bone marrow cells

• The study stated that the capsaicin did this by stopping the cell in the G0-G1 phase and allowing the cell to go through apoptosis

• Leukemic cells were exposed to varying levels of capsaicin (0 µM-1000µM) over a 24 hour period in addition to subjecting cells to a dose of 50 µM for 0 to 120 hours

• These tests suggested that the cells responded to capsaicin in a dose/time dependent manner

• The higher/longer the dose of capsaicin, the faster the cells went through apoptosis

• The study also discovered that when capsaicin was injected into tumors, the tumors would shrink compared to control tumors

• Mice were used as an in vivo test group in both experiments, and were not detrimentally affected by the capsaicin (Ita et al. 2004).

Works CitedIto, Keisuke., Nakazato, Tomonori., Yamato, Kenji, 2004. Induction of Apoptosis in Leukemic Cells by Homovanillic Acid Derivative, Capsaicin, through Oxidative Stress: Implication of Phosphorylation of p53 at Ser-15 Residue by Reactive Oxygen Species. Cancer Research [serial online] 64: 1071-1078.

Mori, Akio., Lehmann, Soren., O’Kelly, James, 2006. Capsaicin, a Component of Red Peppers, Inhibits the Growth of Androgen-Independent, p53 Mutant Prostate. Cancer Research [serial online] 66: 3222-3229.

Methods

HypothesisH0:Treating MCF-7 breast cancer cells with Capsaicin will not cause cell death. H1: Treating MCF-7 breast cancer cells with Capsaicin will induce cell death through apoptosis.

Chili Peppers: A Cure for Breast Cancer? Treating MCF-7 Breast Cancer Cells with Capsaicin at Varying Concentrations

Andrew Sutton Department of Biology, York College of Pennsylvania

Conclusion

• Cultured MCF-7 cells with DMEM + 10% Fetal Bovine Serum + 1% Penstrep + 1% GlutaMAX

• Allowed cells to reach confluency• Removed cells from 30 mL flask using trypsin• Counted cells using a hemocytometer, diluted to 10^6 cells/mL• 150 µL of cells were put into each well of 96 well microtiter plate• Cells were allowed a day to attach and multiply• After first day, cells were administered doses of Capsaicin varying from 0µM to 1000 µM• Capsaicin was dissolved in 100% EtOH or DMSO• Equivalent concentrations of solvent were tested• Four replicates of each concentration were tested• Control healthy cells received 50 µL of growth media• Control dead cells received 50 µL of cyclohexamide or NaN3

• Cell death measured using MTS/PMS assay• Absorbance was measured, the greater the absorbance the more living cells were present• Also ran a Caspase 3 assay to determine if cell death was from apoptosis or necrosis• The higher the fluorescence, the more Caspase 3 protein present suggesting the cells went through apoptosis

Concentration Amt. of Capsaicin/Et

OH (uL)

Amt. of Media (uL)

Total Amt. of Capsaicin/EtOH

(uL)

Total Amt. of Media (uL)

0 uM 0 50 0 2501 uM 0.01 49.99 .05 249.9510 uM 0.1 49.9 0.5 249.550 uM 0.5 49.5 2.5 247.5100 uM 1 49 5 245200 uM 2 48 10 240500 uM 5 45 25 225750 uM 7.5 42.5 37.5 212.51000 uM 10 40 50 200

Results

• Capsaicin was able to kill the MCF-7 breast cancer cells in a dose dependent manner• Control cells were minimally affected by the solvent• Caspase 3 Assay suggests apoptosis as mode of cell death

Future Studies• Does Capsaicin effect all cancers the same way?• Does Capsaicin effect bacteria?

AcknowledgementsThank you Dr. Thompson for all your guidance on this project. Thank you to Joan Carpenter for obtaining the MCF-7 cells

http://en.wikipedia.org/wiki/File:Kapsaicyna.svg

Figure 1. MCF-7 cells in culture dishes prior to various Capsaicin treatments for Caspase 3 Assay.

Figure 2. MCF-7 cells growing and dividing at 400X magnification (left) and 200X (right). Figure 3: MCF-7 cells after treatment with Capsaicin at varying concentrations.

0 mM 1 mMIncreasing concentration

Figure 5. Caspase 3 Assay of Varying Levels of Capsaicin Treated MCF-7 cells after 24 hours. 500 μM Capsaicin treatment showed a significant difference according to an unpaired t-test. P-value <0.05

nsnsns *

Figure 4. Treatment of MCF-7 Cells with Capsaicin at various mM concentrations