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Stetson University

Verified 501(c)(3) Non-Profit

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Project Description
The American Cancer Society estimates in 2014, there were over 1.6 million total new cancer cases in the United States. As cancer continues to be a disease of high incidence, contributing to over 500,000 cancer-related deaths per year in the United States, the need for new cancer therapies is urgent. Natural plants have been used in alternative medicine for generations. Plant-based substances have been reported to have anticancer properties and cause cell death in certain types of cancer like prostate cancer and ovarian cancer. However, the ability of plant-based compounds to induce cell death in human lung cancer cells and human leukemia cells has not been fully explored. The goal of this undergraduate research project is to examine the ability of naturally occurring, plant-based compounds to induce regulated cell death in human lung cancer and leukemia cells.


Biography
Faculty Research Mentor
Dr. Roslyn N. Crowder


The Faculty Research Mentor for the research project is Dr. Roslyn N. Crowder. She has eleven years of experience studying cell death pathways in normal and cancer cells. Her research focuses on investigating agents that selectively cause cell death in cancer cells while leaving normal cells unharmed. Dr. Crowder holds a Ph.D. degree in Microbiology and Immunology from the University of Alabama at Birmingham. Dr. Crowder is a second-year faculty member in the Department of Biology at Stetson University. Her previous research training includes postdoctoral research positions at the University of Pennsylvania and Pennsylvania State University.


Undergraduate Research Students
Sheree Carter


Hometown: Washington, DC
Classification: Senior Career plans after graduation: Attend a post-baccalaureate program Long-term career goals: Become a pediatric oncologist


Ernest Phillips


Hometown: St. Thomas, U.S. Virgin Islands
Classification: Senior
Career plans after graduation: Attend medical school
Long-term career goals: Become a physician


Lauryn Mohler


Hometown: Deltona, FL
Classification: Senior
Career plans after graduation: Attend dental school at the University of Florida
Long-term career goals: Would like to practice dentistry in the Central Florida area specializing in IV sedation and implantology


Jordan Cockfield


Hometown: Wellington, FL
Classification: Junior
Career plans after graduation: Obtain a Ph.D. in Cancer Biology
Long-term career goals: Become a research scientist specializing in cancer biology


Kenneth Massa


Hometown: New Egypt, New Jersey
Classification: Junior
Career plans after graduation: Attend medical school
Long-term career goals: To specialize in anesthesiology


Naser Mubarak


Hometown: Ormond Beach, FL
Classification: Junior
Career plans after graduation: Attend medical school
Long-term career goals: Become a cardiologist


Shelby Curren


Hometown: Atlanta, GA
Classification: Junior
Career plans after graduation: Attend medical school
Long-term career goals: Become a pathologist


Ricardo Vallejo-Calzada


Hometown: Guaynabo, Puerto Rico
Classification: Junior
Career plans after graduation: Attend medical school
Long-term career goals: Become a physician specializing in sports medicine


Kate Ellis


Hometown: Tampa, Florida
Classification: Sophomore
Career plans after graduation: Attend medical school
Long-term career goals: To become a board certified emergency medicine surgeon


Expected project outcomes
After students have performed their experiments and completed their research projects, data will be compiled into student research posters that will be presented at the annual Stetson Showcase that highlights undergraduate student research. Students will also present their
research at the Florida Undergraduate Research Conference (FURC). Additionally, research projects that have generated enough data will be compiled into manuscripts that will be submitted for peer-review. Potential journals to submit student authored manuscripts include Cell Cycle, Journal of Nutritional Biochemistry and Nutrition and Cancer.


Will any products of the project be made available to donors?
Yearly progress reports detailing student research highlights such as oral and posters presentations given and local, regional and national conferences attended, will be compiled and made available to donors.


Methodology
Cell Culture
The research will be conducted by Stetson University undergraduate research students. The H460 human lung cancer cell line and Jurkat Leukemia cell line will be obtained from the American Type Culture Collection (ATCC). H460 and Jurkat cells will be grown in RPMI media supplemented with Penicillin and Streptomycin antibiotics and 10% fetal calf serum. Cells will be maintained in a tissue culture incubator at 37°C with 5% CO2.


Cell Viability
Cell viability will be examined using a commercially available Dead Cell Kit (Millipore) according to manufacturer instructions. Cells will be seeded in a 6-well plate in a 1000μL volume and treated with the soy-based substance for the desired amount of time. Cells will be collected after treatment. 100μL of Muse Annexin V & Dead Cell Reagent will be added to 100μL of cell suspension. The mixture will sit for 20 minutes at room temperature before samples are analyzed on a Muse Cell Analyzer.


Western Blot
Cells will be seeded in 60mm dishes and treated with the soy-based substance for the desired amount of time. Cells will be collected and lysed in lysis buffer (1% NP-40, 10% glycerol, 50mM TRIS, 150mM NaCl, 2mM EDTA) and cleared by centrifugation. Cellular proteins will be resolved on polyacrylamide gels and transferred onto PVDF membranes (Millipore). Membranes will be blocked with 4% non-fat milk before adding primary antibodies. Caspase-8, caspase-3 or PARP primary antibodies will be added to the blot overnight. Blots will be washed three times and then horseradish peroxidase conjugated anti-rabbit or anti-mouse secondary antibodies will be added to the blot. The blots will be washed three more times. ECL solution will be added to visualize the proteins. The blots will be imaged on a GE Healthcare Life Sciences Typhoon Imager.


Timetable
This research will be conducted over a 12 month period beginning in May of 2015. Senior research students, who have previous lab experience, will begin to train junior and sophomore research students in May of 2015.


Budget
General Supplies: $1250
Explanation: Research supplies will be needed to conduct cell death studies on human cancer cells. Research supplies include cell media and supplements to grow and maintain the human lung cancer cells lines. Commercially-available kits used to confirm cell death such as Millipore Muse Count & Viability Kit and Muse Annexin V and Dead Cell Kit will also need to be purchased. Lastly, reagents needed to perform Western blot analysis to investigate the presence and activation of proteins involved in cell death will also need to be purchased.


Detailed Budget: List of Research Supplies



Matching funds: Stetson University Departmental Funds-$675 for lab research supplies


TOTAL amount to raise on CREU: $1,250


How will any additional funds be used?
Additional funds will be used to purchase research lab supplies.

CREU recomends this project at the recommended level

Reviewer comments:

"A description of the soy-based substance and rationale for choosing that substance over others would be helpful."

" She needs to dramatically narrow down to what she will be looking at."

" The research itself would provide good cellular culture testing for undergraduate students and the budget appears to be appropriate."

 

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