Introduction

1. 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