Ross School - Senior Project 2007-08
Mentor: Hugh Mcguinness
Title: The Effects of Beta Carotene on Agrobacterium tumefaciens.
For my senior project, I attempted to test the hypothesis that beta carotene has a preventative effect on the formation of crown gall disease in sunflower plants (Helianthum Dwarf Sunflower). Using greenhouse grown plants, I used a block design to look at whether beta-carotene is more effective as a preventative (plants were given beta carotene before being inoculated) or if it is more effective as a post-preventative measure (plants given beta carotene after exposure to the disease). After the experiment was finished, I completed an in-depth analysis and research paper on the topic.
In 2006, my grandpa was diagnosed with stage 4 pancreatic cancer. We were told that it was terminal, and he was given about 6 months left to live. 2 years later the cancer has disappeared without an explanation and he looks better than ever. From the moment my grandpa was diagnosed, I became interested in the medical field. So, in 10th grade, when we were asked to find an experiment to emulate over the course of 2 months, I began to search for experiments having to do with cancer. This is when I came across 2 or 3 science fair projects that seeked to find the effects of beta-carotene on Agrobacterium tumefaciens. Due to time restrictions, I was not able to pursue this that year, but come senior project time, I knew that this was something I was still interested in.
After researching, I found that beta carotene is what the medical world refers to as a chemopreventative. After several conversations with an oncologist at harvard, I learned of three successful experiments, where beta carotene was used to prevent or suppress cancer in humans. In 1993, a study was conducted in China which proved beta carotene to be effective against gastric cancer. In 2007, it was also found to suppress the expression of breast cancer genes and melanoma cells. Like the two students before me, I wanted to see if beta carotene had the same affect on plant cancer as it does on human cancer.
In my original phone call in early June with Stacy from Carolina Biology Supply, I was informed that New York state did not require a permit for delivery of the Bacterium. With this in mind, it wasn't until July that I placed an order. A few days later I received an e-mail saying that I did need the permit. This was the first road bump in an extremely bumpy experiment. Immediately, I gathered up all the information I needed in order to apply for a permit. With Barbara Jenkins, the head of the department, on my speed dial for the entire summer, the permit was finally approved in the first weeks of September. This loss of time didn't quite fit into my original plan.
I then hit my next problem. Because the heat of the summer was dwindling, we needed to find a warm place with vibrant sunlight to store and grow my seedlings. After many generous offers, we decided that the ECCO farm would be the most convenient choice and I soon began my process. During our time searching for appropriate housing of my plants, I began collecting materials that I needed. Soil and Beta Carotene were easy to find, however sunflowers were a little more difficult. I mistakingly thought that sunflowers were just one of those things that were always available in the hardware store; this was not true. Apparently the very week I set out to gather sunflower seeds, all of the local stores had already called their suppliers to pick up any unsold seed packets. I ended up finding a seed supply store on the Internet and ordered them from there.
So, it is now mid September and I am just getting things started. My seeds have all been planted and while my plants are growing I have begun to research. But just how long would it take these plants to grow?
After treating the plants with their appropriate Beta Carotene dosages and watering them every other day for about a month, it was time to move them into a grow house. A grow room that any drug lord around the world would shake their head at. Apparently, like Rome, a grow room isn't built in a day; something that I did not realize. Searching for lamps took a few days and my set up was a little creative. Text books and hot pink string tied held the lamps up and in order to water the plants I had to crawl under the table.
After weeks of trials, the experiment was unsuccessful, and I had to brainstorm new ideas. I wrote an indepth 20 page research paper discussing cancer, beta carotene, crown gall cancer, and my own experiment on the molecular level.
Through this project, I learned an enormous amount about molecular biology, cancer, and how to conduct long-term experiments.
"Alpha-Tocopherol, Beta-Carotene Cancer Prevention
(ATBC) Trial." National Cancer Institute. 22 July 2003. Viewed 23 Nov.
/pressreleases/ATBCfollowup>. This website provided information on the ATBC trial. This trial determined the long-term effects of vitamin supplemnts in smokers.
Anonymous, The Alpha-Tocopherol, Beta Carotene Cancer Prevention Study Group. 1994. The effect of vitamin E and beta carotene on the incidence of lung cancer and other cancers in male smokers. N Engl J Med. 330:1029–1035.
"Beta-Carotene." Medline Plus. 01 Aug. 2006. 10 Dec. 2007 <http://www.nlm.nih.gov/medlineplus/druginfo/natural/patient-betacarotene.html>. Information on the tested results of Beta-Carotene on various cancers and diseases.
"Beta-Carotene Supplements Confirmed as Harmful to
Those At Risk for Lung Cancer." National Cancer Institute. 13 Dec. 2004.
Viewed 15 Nov. 2007 <http://www.cancer.gov/
clinicaltrials/results/final-CARET1204>. Proof and Trials of Beta-Carotene being harmful on Lung-cancer patients and those who smoke.
"Beta Carotene (Vitamin a)." Molecular Expressions. 4 Mar. 2004. Viewed 15 Nov. 2007 <http://micro.magnet.fsu.edu/vitamins/pages/carotene.html>. Information on Beta carotene, as well as an image of a crystalized Beta carotene cell
Binns, A. N., Sciaky, D., and Wood, H. N. 1982. Variation in hormone autonomy and regeneration potential of cells transformed by strain A66 of Agrobacterium tumefaciens. Cell 31: 605-612.
Bjelakovic, G, et al. 2007. Mortality in Randomized Trials of Antioxidant Supplements for Primary and Secondary Prevention: Systematic Review and Meta-analysis. JAMA. 297:842-857.
Blot WJ, Li JY, Taylor PR, Guo W, Dawsey S, Wang GQ, Yand CS, Zheng SF, Gail M, Li GY, et al. 1993. Nutrition intervention trials in Linxian, China: supplementation with specific vitamin/mineral combinations, cancer incidence, and disease-specific mortality in the general population. 15;85(18):1483-92
Braun, A. C. 1943. Studies on tumor inception in the crown-gall disease. Am. J. Bot. 30:674-677.
"Cancer Deaths Declining: Get Cancer Basics." Mayo Clinic. 18 Jan. 2007. Viewed 1 Dec. 2007 <http://www.mayoclinic.com/health/cancer/CA00003>. Information on cancer (what causes cancer, what is cancer, what increases risks/decreases risks,
"Cancer Statistics From NCI/ACS." The Cancer Cure Foundation. 2002. Viewed 3. Nov. 2007 <http://www.cancure.org/statistics.htm>. Cancer Statistics: estimated deaths per year, approx. amount that will be diagnosed this year.
Cubero J, M.C, Martinez P. Llop and M.M. Lopez. 1998. A simple and efficient PCR method for the detection of Agrobacterium tumefaciensin plant tumors.
Cui Y, Lu Z, Bai L, Shi Z, Zhao WE, Zhao B. 2007. beta-Carotene induces apoptosis and up-regulates peroxisome proliferator-activated receptor gamma expression and reactive oxygen species production in MCF-7 cancer cells. Eur J Cancer. 43(17): 2590-601.
Eugene Nester, Milton P. Gordon, Allen Kerr, et al. 2005. Agrobacterium tumefaciens: From Plant Pathology to Biotechnology. 4:6-9:49-51:145-164
Frieling, U. M., Schaumberg, D. A., Kupper, T. S., Muntwyler, J., Hennekens, C. H. 2000. A Randomized, 12-Year Primary-Prevention Trial of Beta Carotene Supplementation for Nonmelanoma Skin Cancer in the Physicians' Health Study. Arch Dermatol 136: 179-184
Goodman GE, Thornquist MD, Balmes J, Cullen MR, Meyskens FL Jr, Omenn GS, et al. 2004. The Beta-carotene and Retinol Efficacy Trial: Incidence of lung cancer and cardiovascular disease mortality during 6-year follow-up after stopping beta-carotene and retinol supplements. J Natl Cancer Inst. 96:1743-50.
Guruvayoorappan C, Kuttan G. 2007. Beta-carotene inhibits tumor-specific angiogenesis by altering the cytokine profile and inhibits the nuclear translocation of transcription factors in B16F-10 melanoma cells. Integr Cancer Ther. 6(3): 258-70.
H. Nagasawa et al., March-April 1991 Suppression by Beta-Carotene-Rich Algae Dunaliella Bardawil of the Progression, But Not Development, of Spontaneous Mammary Tumours in SHN Virgin Mice, Anticancer Res. 11(2); p. 713-7.
Leppala JM, Virtamo J, Fogelholm R, et al. 2000. Vitamin E and beta carotene supplementation in high risk for stroke: a subgroup analysis of the Alpha-Tocopherol, Beta-Carotene Cancer Prevention Study. Arch Neurol. 57(10):1503-1509.
Murakoshi, Michaiki. Interview with Richard a. Passwater, Ph.D.. Health World. Winter 2004. Viewed 15 nov. 2007
Null, Ph.d., Gary. "Beta Carotene." Garynull, Your Guide to Natural Living. Viewed 30 Nov. 2007 <http://www.garynull.com/documents/BetaCarotine.htm>. Information on various aspects of cancer, as well as the effects of beta carotene on various cancer and diseases
Omenn GS, Goodman GE, Thornquist MD, Grizzle J, Rosenstock L, Barnhart S, et al. 1994. The beta-carotene and retinol efficacy trial (CARET) for chemoprevention of lung cancer in high risk populations: Smokers and asbestos-exposed workers. Cancer Res. 54:2038s-43s.
Omiyama S et Al. 1985. Synergistic combination therapy of 5-fluorouracil, vitamin A, and cobalt-60 radiation for head and neck tumors - antitumor combination therapy with vitamin A. Auris Nasus Larynx. 12(Suppl 2): S239-43.
Ophardt, Charles E. "Beta-Carotene and Vitamin A." Virtual Chembook. 2003. Viewed. 24 Dec. 2007 <http://www.elmhurst.edu/~chm/vchembook/532vitaminA.html>. Beta Carotene information; molecular level
Patlak, Margie. "Taming the Crown Gall." National Academy of Sciences. 2003. Viewed Nov-Dec 2007 <http://www.beyonddiscovery.org/content/view.page.asp?I=187>. Overview and time line of Crown Gall Cancer
"A Physiological Basis for Autonomous Growth of the
Crown-Gall Tumor Cel." PNAS. Apr. 1958. Viewd Oct.-Nov. 2007 <http://www.pubmedcentral.nih.gov/
pagerender.fcgi?artid=335424&pageindex=2#page>. Information on the growth of tumors in plants and animals; similarites and differences
Ray, Amitabha. E-Mail interview. 14 Dec. 2007.
Rencic A, Angert ER, Winans SC. 2005. Unwounded plants elicit Agobacterium vir induction and T-DNA transfer: transformed plant cells produce opines yet are tumor free. Mol Microbiol. 57:1522–1531. doi: 10.1111/j.1365-2958.2005.04763.x. [PubMed]
Richard A. Passwater, Ph.D., 1984. Beta-Carotene: The Backstage Nutnent Now Universally Recognized for Cancer Prevention, Keats Publishing Inc., New Canaan, Conn., 30:18
Santamaria LA and Santamaria AB. 1990. Cancer chemoprevention by supplemental carotenoids and synergism with retinol in mastodynia treatment. Med Oncol Tumor Pharmacother; 7(2-3): 153-67.
Schaumberg DA, Frieling UM, Rifai N, et al. 2004. No effect of beta-carotene supplementation on risk of nonmelanoma skin cancer among men with low baseline plasma beta-carotene. Cancer Epidemiol Biomarkers Prev. 13(6):1079-1080
S.K. Gaby and V.N. Singh, 1991. "B-Carotene," Vitamin Intake and Health: A Scientific Review, S.K. Gaby, A. Bendich, V. Singh and L. Macklin (eds.) Marcel Dekker, N.Y. p.30.
Toma S, Bonelli L, Sartoris A, et al. 2003. beta-carotene supplementation in patients radically treated for stage I-II head and neck cancer: results of a randomized trial. Oncol Rep. 10(6):1895-1901
Tornwall ME, Virtamo J, Haukka JK, et al. 2001. Alpha-tocopherol (vitamin E) and beta-carotene supplementation does not affect the risk for large abdominal aortic aneurysm in a controlled trial. Atherosclerosis; 157(1):167-173.
Community Member (Details)
Dr. Ann Aguanno
Molecular Biologist at Mary Mount Manahatten
Helpful in Understanding A. Tumefaciens on the molecular level
Dr. Ray Abhitbah
Provided me with documents from past experiments where beta carotene was effective against human cancers
Understanding Molecular Biology and provided me ways to make my project for indepth on the molecular level.