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Anti-Proliferative Effects of <I>Garcinia</I>

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Anti-Proliferative Effects of <I>Garcinia</I> City University of New York (CUNY) CUNY Academic Works All Dissertations, Theses, and Capstone Projects Dissertations, Theses, and Capstone Projects 2-2018 Anti-Proliferative Effects of Garcinia Fruits in Breast Cancer Cells Harini Anandhi Senthilkumar The Graduate Center, City University of New York How does access to this work benefit ou?y Let us know! More information about this work at: https://academicworks.cuny.edu/gc_etds/2549 Discover additional works at: https://academicworks.cuny.edu This work is made publicly available by the City University of New York (CUNY). Contact: [email protected] ANTI-PROLIFERATIVE EFFECTS OF GARCINIA FRUITS IN BREAST CANCER CELLS Harini Anandhi Senthilkumar A dissertation submitted to the graduate faculty in Biology towards partial fulfillment of the requirements for the degree of Doctor of philosophy, The City University of New York 2018 © 2018 HARINI ANANDHI SENTHILKUMAR All rights reserved ii Antiproliferative effects of Garcinia fruits in breast cancer cells by Harini Anandhi Senthilkumar This manuscript has been read and accepted for the Graduate faculty in Biology in satisfaction of the dissertation requirement for the degree of Doctor of Philosophy. Date Dr. Edward Kennelly Chair of Examining Committee Date Dr. Cathy Savage-Dunn Executive Officer Supervisory Committee: Dr. Jimmie Fata Dr. Moira Sauane Dr. Prabodhika Mallikaratchy Dr. Kurt Reynertson THE CITY UNIVERSITY OF NEW YORK iii ABSTRACT Antiproliferative effects of Garcinia fruits in breast cancer cells by Harini Anandhi Senthilkumar Advisor: Dr. Edward J. Kennelly Breast cancer continues to be the second leading cause of death in women, even with the recent advances in research that has led to an improved understanding of molecular pathways and targets. Increasing mortality due to recurrence and lack of targeted drugs, especially in aggressive breast cancer subtypes such as triple negative breast cancer, has gained attention from the research community. With nearly 50% of the commonly used cancer drugs being sourced from plants, bioactive small molecules derived from a natural origin have immense potential as therapeutics. This study focuses on the anti-proliferative activities of benzophenones isolated from the edible fruits of Garcinia xanthochymus and Garcinia paucinervis in breast cancer cells, particularly triple negative breast cancer cells. Novel benzophenones paucinones E – I from Garcinia paucinervis were tested on a panel of breast cancer cell lines MDA-MB-231, MCF-7 and SKBR-3 with varying receptor statuses. Paucinone H was active across the panel of cell lines studied, with IC50 values ranging from 10 to 20 µM. Since the activity was significant on MDA- MB-231, the possible pathways in MDA-MB-231 that paucinone H could modulate were iv investigated. Paucinone H induced apoptosis in a dose dependent fashion at IC50 concentration and decreased mitosis and migration at half IC50 concentration in MDA-MB- 231. In addition to the research described above, a comprehensive review of dietary phytoestrogens and their impact on breast pathophysiology is also presented in Chapter 2. Appendices A and B discuss the use of UPLC-QTOF-MS based metabolomics to verify bioactivities of various vegetative organs of Garcinia oblongiflia and the identification of novel benzophenones from Garcinia paucinervis v ACKNOWLEDGEMENTS My experience in Graduate school has been equally exciting and challenging. It started out because of my love for Science and my inquisitiveness in understanding the biology of Cancer. After my exposure to natural products in the Phytochemistry course during my first year in school, I developed interest in analyzing the potential use of natural compounds in the prevention and treatment of cancer. I am thankful to Dr. Kennelly and Dr. Fata for their support in helping me carry out my research. There is no achievement that comes without hard work and support of people who believe in you. In my case, I am grateful to my husband who always stood with me and was my constant source of strength and support through this long journey. I am also grateful to my parents who always believed in me. There were times when my mother’s words have helped me realize my dreams and to overcome challenges. I am thankful to my father for his calm and consoling words. With his professional experience as an Engineer in Research and development of energy efficient automobile engines, he could well relate to the frustrations in my failed experiments. Together we have shared our frustrations and love for Science. I am also thankful to my sister who has cheered me up when I needed a laugh. I would like to thank my lab mates and good friends Kaushiki Chatterjee, Vanya Petrova, Adam Negrin and Taylan Morcol for all the technical and non-technical small talks that made my time in lab an enriching experience. I am also thankful to my colleague Ping Li, who was an excellent professional to work with as a co-author. Appendices A and B of this thesis have been published in peer reviewed journals (citations listed below). Chapter 2.1 is a manuscript under review. At the time of completion of this thesis, the second set of reviewer comments have been addressed. vi Li P, Anandhi Senthilkumar H, Figueroa M, Wu SB, Fata JE, Kennelly EJ, Long C 2016. UPLC-QTOFMS(E)-guided dereplication of the endangered Chinese species Garcinia paucinervis to identify additional benzophenone derivatives. J Nat Prod 79: 1619-1627. Li P, AnandhiSenthilkumar H, Wu SB, Liu B, Guo ZY, Fata JE, Kennelly EJ, Long CL 2016. Comparative UPLC-QTOF-MS-based metabolomics and bioactivities analyses of Garcinia oblongifolia. J Chromatogr B Analyt Technol Biomed Life Sci 1011: 179-195 A good research starts with an exciting idea. But thoughtful guidance is the one that leads to achievement of best results. I am thankful to my committee members Dr. Prabodhika Mallikaratchy, Dr. Kurt Reynertson and Dr. Moira Sauane for their time, guidance and constantly looking out for the best for my career. Their advice has been critical in shaping my research and in educating myself on ways to overcome technical obstacles throughout the course of research and my time in Graduate school. Finally, I would like to thank my angel son Mukhil whose little feet I never got to touch. Although his life was short, his eternal presence in my heart will be my greatest source of strength for now and forever. vii TABLE OF CONTENTS Abstract .................................................................................................................................. iv Acknowledgements ................................................................................................................ vi List of Figures ........................................................................................................................ x List of Tables ......................................................................................................................... xii List of Abbreviations ............................................................................................................. xii Chapter 1: Introduction ....................................................................................................... 1 1.1 Garcinia xanthochymus and Garcinia paucinervis fruits – Phytochemistry ................................................................................................................ 4 Chapter 2: Breast cancer and bioactive compounds from plants .................................. 13 *2.1 Dietary phytoestrogens and their impact on breast biology ......................................... 14 2.2 Breast cancer pathways modulated by compounds from Garcinia ............................................................................................................................... 39 2.3 Triple negative breast cancer – Introduction ................................................................. 46 *Chapter 3: Molecular mechanisms targeted by benzophenones from Garcinia paucinervis against breast cancer cell lines with varying receptor statuses ................. 51 3.1 Materials and Methods ................................................................................................. 53 viii 3.2 Results ........................................................................................................................ 61 3.3 Discussion .................................................................................................................. 70 Chapter 4: Conclusion ................................................................................................... 72 Chapter 5: Future studies ............................................................................................. 75 *Appendix A: Comparative UPLC-QTOF-MS-based metabolomics and bioactivities analyses of Garcinia oblongifolia ..................................................... 79 *Appendix B: UPLC-QTOF-MS guided dereplication of an endangered Chinese Garcinia species to identify cytotoxic benzophenones in triple negative and other breast cancer cell lines ..................................................................82 Chapter 6: References .................................................................................................. 86 *Results that have been published in and submitted to peer reviewed journals ix List of Figures Page Figure 1: Garcinia paucinervis fruits ............................................................................. 3 Figure 2: Garcinia xanthochymus
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