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Targeting LEDGF/P75 to Sensitize Chemoresistant Prostate Cancer Cells to Taxanes Leslimar Rios-Colón

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Targeting LEDGF/P75 to Sensitize Chemoresistant Prostate Cancer Cells to Taxanes Leslimar Rios-Colón Loma Linda University TheScholarsRepository@LLU: Digital Archive of Research, Scholarship & Creative Works Loma Linda University Electronic Theses, Dissertations & Projects 6-2017 Targeting LEDGF/p75 to Sensitize Chemoresistant Prostate Cancer Cells to Taxanes Leslimar Rios-Colón Follow this and additional works at: http://scholarsrepository.llu.edu/etd Part of the Medical Biochemistry Commons, Medical Pharmacology Commons, and the Oncology Commons Recommended Citation Rios-Colón, Leslimar, "Targeting LEDGF/p75 to Sensitize Chemoresistant Prostate Cancer Cells to Taxanes" (2017). Loma Linda University Electronic Theses, Dissertations & Projects. 459. http://scholarsrepository.llu.edu/etd/459 This Dissertation is brought to you for free and open access by TheScholarsRepository@LLU: Digital Archive of Research, Scholarship & Creative Works. It has been accepted for inclusion in Loma Linda University Electronic Theses, Dissertations & Projects by an authorized administrator of TheScholarsRepository@LLU: Digital Archive of Research, Scholarship & Creative Works. For more information, please contact [email protected]. LOMA LINDA UNIVERSITY School of Medicine in conjunction with the Faculty of Graduate Studies ____________________ Targeting LEDGF/p75 to Sensitize Chemoresistant Prostate Cancer Cells to Taxanes by Leslimar Ríos-Colón ____________________ A Dissertation submitted in partial satisfaction of the requirements for the degree Doctor of Philosophy in Pharmacology ____________________ June 2017 © 2017 Leslimar Ríos-Colón All Rights Reserved Each person whose signature appears below certifies that this dissertation in his/her opinion is adequate, in scope and quality, as a dissertation for the degree Doctor of Philosophy. _______________________________________________________________________ John Buchholz, Professor of Pharmacology _______________________________________________________________________ Eileen Brantley, Assistant Professor of Pharmacology _______________________________________________________________________ Carlos A. Casiano, Professor of Microbiology and Molecular Genetics _______________________________________________________________________ Daisy De León, Professor of Physiology _______________________________________________________________________ Nouri Neamati, Professor of Medicinal Chemistry, The University of Michigan _______________________________________________________________________ Kimberly J. Payne, Associate Professor of Pathology and Human Anatomy _______________________________________________________________________ Lubo Zhang, Professor of Pharmacology iii ACKNOWLEDGEMENTS I am extremely grateful of my advisor Dr. Carlos A. Casiano, an exemplary scientist, who dedicated time and effort so that I could evolve from a student to a detailed-oriented researcher. I am thankful for your trust and guidance and for believing that not only I had the potential to obtain this degree, but that I can make an impact in this field. I am also thankful to all the past and current members of my laboratory. Especially I want to thank Christina K. Du Ross and Leanne Woods-Burnham, who have become my family. We have gone through so much together! I will always remember the singing, laughter, crying and the many scientific debates we had. I could not have asked for two better people to share this journey with. I am also thankful to Dr. Anamika Basu, who was an incredible source of knowledge not only for scientific topics, but also of how to manage life as a graduate student. I also want to thank Dr. Tino W. Sanchez for all the stimulating scientific conversations and knowledge provided. I also want to thank my two former summer students, Catherine Elix and Ivana Alicea Polanco. It was an honor to be able to help you navigate the complex world of research! I could not have been able to complete this without the love and support of my family. To my husband Edwin Pagán, who never let go of my hand through this path no matter how demanding it was. To my parents, Lester Ríos and Margarita Colón, who were my biggest supporters since the day I decided to travel many miles in order to achieve my dreams. To my dearest Adeline, you are my biggest accomplishment and the reason I want to improve everyday as a scientist and as a human being. iv I also want to acknowledge my undergraduate research mentor, Dr. Adriana Báez. I appreciate all the support and for always giving me advice. And finally, I would like to thank God who always has guided me through mysterious paths with his angels guiding the way. v CONTENT Approval Page .................................................................................................................... iii Acknowledgements ............................................................................................................ iv List of Figures .................................................................................................................... ix List of Tables ..................................................................................................................... xi List of Abbreviations ........................................................................................................ xii Abstract ............................................................................................................................ xvi Chapter 1. Introduction ..............................................................................................................1 Prostate Cancer and Available Therapies .........................................................1 Inflammation and Prostate Cancer .....................................................................6 The Lens Epithelium-Derived Growth Factor of 75kD .....................................6 LEDGF Structure and Function .........................................................................7 Cellular Protective and Survival Functions of LEDGF/p75 ............................13 Target Genes of LEDGF/p75 ...........................................................................14 Interacting Partners of LEDGF/p75 .................................................................16 Regulation of LEDGF/p75 Expression and Function .....................................17 Autoantibody Responses to LEDGF/p75 in Cancer and Inflammatory Conditions .........................................................................20 Role of LEDGF/p75 in Leukemia ....................................................................23 Overexpression and Role of LEDGF/p75 in Solid Tumors ............................26 LEDGF/p75 Promotes Chemoresistance in Cancer Cells ...............................28 Targeting LEDGF/p75 and its Potential in Combinatorial Therapy ................30 Repurposing HIV-Based LEDGF/p75 Inhibitors for Cancer Treatment ........34 Summary Highlights .......................................................................................37 Hypothesis and Purpose of Dissertation Work ................................................40 References ........................................................................................................41 2. Targeting the Stress Oncoproten LEDGF/p75 to sensitize Chemoresistance Prostate Cancer Cells to Taxanes ..............................................57 Abstract ............................................................................................................58 Introduction ......................................................................................................59 Results ..............................................................................................................61 vi LEDGF/p75 is Overexpressed in DTX-Resistant DU145 and PC3 Cells ...........................................................................................................61 DU145-DR and PC3-DR Cells are Resistant to Multiple Taxanes but not to TRAIL .......................................................................................66 LEDGF/p75 Depletion Sensitizes DTX-resistant PCa cells to Clinically Relevant Taxanes .....................................................................70 LEDGF/p75 Retains Structural Integrity During Taxane-Induced Cell Death but is Robustly Cleaved During TRAIL-Induced Apoptosis ...................................................................................................76 LEDGF/p75 Depletion Does Not Influence the Expression of the Multidrug Resistance Protein P-glycoprotein in DTX-Resistant PCa Cells ...................................................................................................81 Discussion ........................................................................................................83 Materials and Methods ....................................................................................91 Cell lines, Antibodies and Reagents .........................................................91 Viability Assays ........................................................................................92 Quantitative Real-time PCR ......................................................................92 Immunoblotting Procedures .......................................................................93 Indirect Immunofluorescence Microscopy ................................................94 RNA Interference-Mediated Knockdown of LEDGF/p75 in PCa cells ..........................................................................94 Clonogenic Assays .....................................................................................95
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