INVESTIGATIONS OF ANTI-CANCER COMPOUNDS FROM ROOT EXTRACTS OF ECHINACEA

1. INVESTIGATIONS OF ANTI-CANCER COMPOUNDS FROM ROOT EXTRACTS OF ECHINACEA Lakeshia Wright, Advisors, Todd Gary and E. Lewis Myles

2. Introduction: Medicinal Plants • Plants are the source of medicine. 25% of prescriptions in U.S. are plant-based. • Herbal products become increasingly important to the international health care.

3. Medicinal Plants • Taxol(paclitaxel) is a naturally occuring substance in evergreen YEW. • It is an anticancer drug that inhibits cell division. • Echinacea is a genus native to North. It has been utilized as a healing herb by Native American people.

4. Echinacea/Purple Cone Flower

5. SECONDARY METABOLITES • Primary Metabolites are compound synthesized by plants and are needed for growth and development. Primary Compounds are need for survival (proteins, carbohydrate, lipids and Nucleic Acids) • Secondary Metabolites are compounds that are not used by the plant for growth and development. The primary role of secondary compounds are for protection against environmental stress.

6. MEDICINAL PLANT COMPOUNDS • PHENOLICS: Flavonoids (Anti-cancer) , Digitalin (Heart Conditions) • Alakaloids: Strychnine (toxic poison), Morphine (analgesic), codeine (cough supressant), atropine (neurotoxin inhibitor). • Terpenoids: menthol, mints • Other compouds such as Asprin, Cumarin (inhibits blood cloting)

8. Oxidants • Free radicals are important and essential as intermediates in primary metabolism and the regulation of specific signal transduction pathways and gene expression. • A variety of oxidants, including superoxide, HP, lipid hydroperoxides, and hydroxyl radical, are produced by diverse initiating agents or modalities such as ionizing radiation, redox cycling drugs, leakage from the mitochondria during electron transport, activation of signal transduction pathways, and a variety of cell types of the immune system when stressed or activated. • oxidizing species can damage cells and tissues

9. Oxidants • Fig. 1. A, hierarchical clustering of RNA expression observed for HP, MEN, and TBH treatment of MCF7 cells using Pearson correlation and complete linkage. Average expressions of the 3 replicates of 446 genes are shown in logarithmic scale. All of the expressions shown are relative to untreated cells. For each treatment, expressions shown are for 0, 1, 3, 7, and 24 h after treatment (left to right). Selected subclusters (A–F) are shown on the right with clone identification numbers (bold text, IMAGE clones; regular text, Incyte clones) and gene symbols. B, MDS plot. The MDS plot (of the total 446 genes shown in A) illustrates the relationship among all of the samples; MEN (blue), HP (red), and TBH (green). Labeled in black, the initial time point (0 h or untreated) for all three treatments. The average expression ratios of three replicates were used in the calculation of the Pearson correlation

10. Materials & Methods • Extraction was performed using 2 grams of Echinacea to 4ml acetone . • The suspension was centrifuged at 15xg. • Extract was collected and experimentally used on a culture of BT549 breast cancer cells • Cells were counted and viability determined by hemocytometer.

11. Materials & Methods • Six well plates were used to culture 300,000 cells. • Three samples were examined and compared (DMSO and extract + DMSO). 4 to 33 ug of each were added. • Cells were treated for 4 days and cell proliferation, viability index was determined.

12. Materials and Methods • The cells were exposed to the extract for 4 days. • The cells were detached from the flask with trypsin. • The cells were counted using a Hemocytometer • Results were recorded

13. Echinacea Pallida

16. Inhibition of the GADD family of genes protects FO-1 melanoma cells from Ad.mda-7-mediated cell death.

18. Future Research • We will be able to tell if oxidants are increased or decreased to cause the death of cancer cells • Whether or not the use of signal transduction along with oxidants will increase the chances of apoptosis

19. References • Chuang, Yao-Yu Eric; Gene Expression after Treatment with Hydrogen Peroxide, Menadione, or t-Butyl Hydroperoxide in Breast Cancer Cells Radiation Biology Branch, Center for Cancer Research, National Cancer Institute [Y-Y. E. C., G. V. R. C., J. A. C., D. C., M-H. T., W. D., H. Y., S. Z., A. R., J. B. M.], and Cancer Genetics Branch, National Human Genome Research Institute [Y. C.], NIH, Bethesda, Maryland 20892, and Genome Institute of Singapore, Singapore 117528 [E. T. L.] • Sarosi, Luis Alberto Gomez; Hydrogen peroxide increases a 55-kDa tyrosinase concomitantly with induction of p53-dependent p21 waf1 expression and a greater Bax/Bcl-2 ratio in pigmented melanoma*1 ; Tumor Cell Biology Laboratory, Centre of Microbiology and Cell Biology, IVIC, Apartado 21827, Caracas 1020 A, Venezuela Received 22 October 2003.