Chili Peppers: A Cure for Breast Cancer?
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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. http://en.wikipedia.org/wiki/File:Kapsaicyna.svg. Introduction. Methods.

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Introduction 7089222

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

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

Introduction

Methods

  • 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

  • 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).

Results

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

Hypothesis

ns

*

ns

ns

H0: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.

Figure 5. Caspase3 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

Conclusion

  • 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

Figure 1. MCF-7 cells in culture dishes prior to various Capsaicin treatments for Caspase3 Assay.

Future Studies

  • Does Capsaicin effect all cancers the same way?

  • Does Capsaicin effect bacteria?

Works Cited

Ito, Keisuke., Nakazato, Tomonori., Yamato, Kenji, 2004. Induction of Apoptosis in Leukemic Cells by HomovanillicAcid 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.

0 mM

Increasing concentration

1 mM

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.

Acknowledgements

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


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