Dandelion Root & Long Pepper Extracts
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University of Windsor Scholarship at UWindsor Electronic Theses and Dissertations 2014 ANTICANCER ACTIVITY OF NATURAL HEALTH PRODUCTS (DANDELION ROOT & LONG PEPPER EXTRACTS); EXTENSIVE STUDY OF EFFICACY AND MECHANISM OF ACTION Pamela Ovadje University of Windsor Follow this and additional works at: http://scholar.uwindsor.ca/etd Recommended Citation Ovadje, Pamela, "ANTICANCER ACTIVITY OF NATURAL HEALTH PRODUCTS (DANDELION ROOT & LONG PEPPER EXTRACTS); EXTENSIVE STUDY OF EFFICACY AND MECHANISM OF ACTION" (2014). Electronic Theses and Dissertations. Paper 5209. This online database contains the full-text of PhD dissertations and Masters’ theses of University of Windsor students from 1954 forward. These documents are made available for personal study and research purposes only, in accordance with the Canadian Copyright Act and the Creative Commons license—CC BY-NC-ND (Attribution, Non-Commercial, No Derivative Works). 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ANTICANCER ACTIVITY OF NATURAL HEALTH PRODUCTS (DANDELION ROOT & LONG PEPPER EXTRACTS); EXTENSIVE STUDY OF EFFICACY AND MECHANISM OF ACTION By PAMELA OVADJE A Dissertation Submitted to the Faculty of Graduate Studies through the Department of Chemistry and Biochemistry in Partial Fulfillment of the Requirements for the Degree of Doctor of Philosophy at the University of Windsor Windsor, Ontario, Canada 2014 © 2014 Pamela Ovadje Anticancer Activity of Natural Health Products (Dandelion Root & Long Pepper Extracts); Extensive Study of the Efficacy and Mechanism of Action by Pamela Ovadje APPROVED BY: __________________________________________________ P. Dou, External Examiner Wayne State University __________________________________________________ A. Swan Department of Biological Sciences __________________________________________________ M. Boffa Department of Chemistry & Biochemistry __________________________________________________ P. Vacratsis Department of Chemistry & Biochemistry __________________________________________________ S. Pandey, Advisor Department of Chemistry & Biochemistry September 11, 2014 DECLARATION OF CO-AUTHORSHIP/PREVIOUS PUBLICATION I. DECLARATION OF CO-AUTHORSHIP I hereby declare that this thesis incorporates materials that are a result of joint research. This thesis includes work carried out in collaboration with Dr. Caroline Hamm, Dr. Jose-Antonio Guerrero and Dr. John Thor Arnason, under the supervision of Dr. Siyaram Pandey. The collaboration with Dr. Hamm is covered in chapters 2 – 6 The collaboration with Dr. Guerrero and Dr. Arnason are covered in chapters 5 – 6. In all cases, the experimental design, execution, data analysis, interpretation and manuscript preparation were performed by the author and all listed authors read and approved the final manuscript, prior to submission. Contributions from no-authors came in form of proofreading manuscripts, and this was covered in the acknowledgements sections of the published work, where appropriate. I am aware of the University of Windsor Senate Policy on Authorship and I certify that I have properly acknowledged the contribution of other researchers to my thesis. I certify that, with the above qualification, this thesis and the research to which it signifies, is the product of my own work. iii II. DECLARATION OF PREVIOUS PUBLICATIONS This thesis incorporates five original papers that have been published or submitted for publication in peer-reviewed journals, listed below; Thesis Chapter Publication Title/Full Citation Publication Status Selective Induction of Apoptosis Through Activation of Caspase-8 In Human Leukemia Cells (Jurkat) By Dandelion Root Extract. Chapter 2 Journal of Ethnopharmacology, 133 (1): 86- Published 91 Selective Induction Of Apoptosis And Autophagy Through Treatment With Chapter 3 Dandelion Root Extract In Human Pancreatic Published Cancer Cells. Pancreas, 41 (7): 1039-1046 Efficient Induction Of Extrinsic Cell Death By Dandelion Root Extract In Human Chronic Chapter 4 Myelomonocytic Leukemia (CMML) Cells, Published PloS one, 7 (2): e30604 Fractionation, Phytochemical & Gene Expression Analysis of the Selective Chapter 5 Anticancer Activity of Dandelion Root Extract In Progress in Human Cancer Cells Evaluation Of The Efficacy & Biochemical Mechanism Of Cell Death Induction By Long Under Pepper Extract Selectively In In-Vitro And In- Chapter 6 Vivo Models Of Human Cancer Cells. PloS Revision one, MS#: PONE-D-14-20706 I certify that I have obtained a written permission from the copyright owner(s) to include the above published material(s) in my thesis. I certify that the above material describes work completed during my registration as graduate student at the University of Windsor. I declare that, to the best of my knowledge, my thesis does not infringe upon anyone’s copyright nor violate any proprietary rights and that any ideas, techniques, quotations, or any other material from the work of other people iv included in my thesis, published or otherwise, are fully acknowledged in accordance with the standard referencing practices. Furthermore, to the extent that I have included copyrighted material that surpasses the bounds of fair dealing within the meaning of the Canada Copyright Act, I certify that I have obtained a written permission from the copyright owner(s) to include such material(s) in my thesis. I declare that this is a true copy of my thesis, including any final revisions, as approved by my thesis committee and the Graduate Studies office, and that this thesis has not been submitted for a higher degree to any other University or Institution. v ABSTRACT The incidence of cancer worldwide is continuously on the rise, and the death toll associated with cancer, is constantly increasing. Available cancer therapy is unable to combat this ever-changing disease, and the severe side effects associated with various forms of therapy, indicate a serious need for the development of more effective and safer alternatives to currently available treatment. The use of natural health products in disease treatment has contributed to the development of over 75% of available chemotherapy. In this thesis, we study the potential anti-cancer effect of two natural health products, dandelion root and long pepper extracts. The major objectives of this work were to; - Evaluate the efficacy and mechanism of these extracts in various cancer models, - Assess any potential safety and toxicity issues associated with the use of these extracts and, - De-convolute and identify the pharmacologically active components that contribute to the anti-cancer activity of these extract Using standard biochemical and morphological assays, the induction of various programmed cell death processes was assessed, following treatment in cancer and non-cancer cell models, as well as in animal models. The results obtained indicate that dandelion root and long pepper extracts were efficacious in selectively inducing apoptosis and pro-death autophagy in various vi cancer cell models. Dandelion root extract rapidly activated the extrinsic pathway of apoptosis, a situation that was crucial to apoptosis induction in leukemia cells, but not required for colorectal cancer cells. Gene expression analysis showed that dandelion root extract efficiently targets multiple pathways to promote its anti-cancer activity. The generation of reactive oxygen species by long pepper extract treatment, appeared to be partially responsible for the induction of apoptosis in cancer cells. Furthermore, the lack of toxicity observed in animal models, on oral administration regimens of dandelion root and long pepper extracts, further confirm the safety of these extracts. Interestingly, the same regimen of oral administration of these extracts was successful in halting the growth of colon tumors in xenograft models. These findings provide scientific validation concerning the safe and effective use of natural health products as non-toxic and potentially more efficacious forms of therapy. vii DEDICATION This work is dedicated to the memory of Mr. Kevin Couvillon, whose fight with Leukemia ended in 2010. You are forever in our hearts and continually inspire us to work harder in the fight against cancer. viii ACKNOWLEDGEMENTS With much gratitude, I would like to acknowledge the efforts of my supervisor, Dr. Siyaram Pandey, for his continued guidance, support and mentorship for the duration of my studies. His enthusiasm and willingness to share his extensive knowledge is always inspiring. His love for mangoes (and other fruits) is a testament to his love for keeping the Pandey Lab healthy. I would also like to acknowledge my committee members, Dr. Michael Boffa, Dr. Otis Vacratsis and Dr. Andrew Swan for all the help and input over the years. Furthermore, I would like to extend my gratitude to all the members of the Chemistry & Biochemistry department, who have made the last few years memorable. To all the members of the Pandey lab, both past and present, I would like to thank you all for all the support, advices, laughs and frustrating times. And to my collaborators, thank you for all the advice and support. All of this support