Marshall University Marshall Digital Scholar Theses, Dissertations and Capstones 2019 Potential Applications of Capsaicinoids in Small Cell Lung Cancer Therapy Jamie Rae Friedman [email protected] Follow this and additional works at: https://mds.marshall.edu/etd Part of the Medical Cell Biology Commons, Medical Pharmacology Commons, and the Oncology Commons Recommended Citation Friedman, Jamie Rae, "Potential Applications of Capsaicinoids in Small Cell Lung Cancer Therapy" (2019). Theses, Dissertations and Capstones. 1226. https://mds.marshall.edu/etd/1226 This Dissertation is brought to you for free and open access by Marshall Digital Scholar. It has been accepted for inclusion in Theses, Dissertations and Capstones by an authorized administrator of Marshall Digital Scholar. For more information, please contact [email protected], [email protected]. POTENTIAL APPLICATIONS OF CAPSAICINOIDS IN SMALL CELL LUNG CANCER THERAPY A dissertation submitted to the Graduate College of Marshall University In partial fulfillment of the requirements for the degree of Doctor of Philosophy In Biomedical Sciences by Jamie Rae Friedman Approved by Dr. Piyali Dasgupta, Committee Chairperson Dr. Monica Valentovic Dr. Gary Rankin Dr. Richard Egleton Dr. Travis Salisbury Marshall University May 2019 © 2019 Jamie Rae Friedman ALL RIGHTS RESERVED iii DEDICATION I would like to dedicate this work to my family (both human and canine) who have supported me throughout all of my studies. To my parents, Steve and Kathy, always told me I could achieve anything I wanted and stayed by my side through the ups and downs. To my brother, Scott, who was always a phone call away to make me laugh and watch sports together even though we were 8 hours away. My parents and brother let me talk about my project often, even though they usually didn’t have a clue what I was talking about. To my Pop-pop and Mom- mom, Stan and Alma, who always were interested in my research, facetiming me almost daily to check in. My aunts and uncles always sent words of encouragement to me, reminding me that eventually I would actually finish. I also can’t forget to mention the Friedman Dogs, Jake, Sosa, and Josie. Josie has worked hard by my side (usually napping) so that she could earn her puppy PhD with me. Lastly, I’d like to dedicate this to my grandparents who were diagnosed with lung cancer and inspired my interest in cancer related research, Pete and Rae. And to my Grammy, Shelia, who I know has been with me through this whole journey. I love you all. iv AKNOWLEDGEMENTS First, I’d like to acknowledge my mentor Piyali Dasgupta. She took a chance on me when she allowed me to join her lab and become her first PhD student. She allowed me to work independently and taught me a lot about what it takes to become a successful scientist. Next, I need to thank my committee members, Monica Valentovic, Gary Rankin, Travis Salisbury, and Richard Egleton. They always had their doors open to me. They all helped me tremendously during my course work as well, always believing in me. Sarah Miles, thank you for helping me with the writing of my dissertation and for dealing with all of my crazy text messages. Thank you all for helping me to become a scientist that I had always wanted to be. Thank you Dr. Valentovic for allowing me to become an unofficial lab member and borrow Katie Brown or Dakota Ward. I do not know what I would have done if I hadn’t been able to run across the hall to them. Working with Katie and Dakota both every day was some of the most fun I could’ve had while “sciencing.” Both of them have become my little lab family and I’m so grateful that they (and their families) took me in. I’m also thankful for being able to work with great people in the lab who knew exactly how to push my buttons, yet always got their work done. Nick, Kate, Stephen, and Justin were the best undergraduates I could have had the chance to work with. Austin Akers was my first lab friend and maintained my friendship throughout my four years. He has made a lasting impact on my time here at Marshall. I also would like to acknowledge my friends outside of the BMS Program, who all have had to bring me back down to reality when I needed it. Lastly, I would like to acknowledge the original ladies I came into this program with, Morghan Getty Collins, Sarah Stevens, and Lexie Blalock. I would never have survived my classes without them and can’t believe we’ve made it this far. Morghan, Sarah, and Lexie are some of the strongest women I know, and I look up to each one v of them. I can’t imagine my time at Marshall without any of those mentioned; thank you so much for helping me along the way. I owe all of my success at Marshall to all of you. vi TABLE OF CONTENTS Dedication ...................................................................................................................................... iv Aknowledgements ........................................................................................................................... v List of Tables ................................................................................................................................ xii List of Figures .............................................................................................................................. xiii Abstract ........................................................................................................................................ xvi Chapter 1: Introduction ................................................................................................................... 1 Lung Cancer and Capsaicin ................................................................................................ 1 Lung cancer ............................................................................................................. 1 Capsaicin ................................................................................................................. 3 Statement of hypothesis .......................................................................................... 4 Chapter 2: Capsaicinoids Enhance Chemosensitivity to Chemotherapeutic Drugs ....................... 6 Abstract ............................................................................................................................... 7 Introduction ......................................................................................................................... 8 Antimetabolites ................................................................................................................. 10 Platinum-Based Drugs ...................................................................................................... 17 Anthracyclines .................................................................................................................. 29 Camptothecin Analogs ...................................................................................................... 35 Targeted Signal Transduction Inhibitors .......................................................................... 42 Radiation Therapy ............................................................................................................. 46 Conclusions and Future Directions ................................................................................... 50 Chapter 3: Capsaicin Synergizes with Camptothecin to Induce Increased Apoptosis in Human Small Cell Lung Cancers Via the Calpain Pathway ..................................................................... 51 vii Abstract ......................................................................................................................................... 52 Introduction ....................................................................................................................... 52 Materials and methods ...................................................................................................... 55 Reagents ................................................................................................................ 55 Cell culture ............................................................................................................ 56 Preparation of lysates ............................................................................................ 56 Measurement of caspase-3 activity ....................................................................... 56 Cell death ELISA .................................................................................................. 57 Chicken chorioallantoic membrane (CAM) assay ................................................ 58 Preparation of tumor lysates from CAM .............................................................. 58 Measurement of calpain activity ........................................................................... 59 Statistical analysis ................................................................................................. 60 Results ............................................................................................................................... 61 A concentration of 10 µM capsaicin does not cause significant apoptosis (p£0.05) in human small cell lung cancer (SCLC) cell lines. .............................................. 61 The combinatorial apoptotic activity of camptothecin and capsaicin is greater than these drugs treated alone in human SCLC cells .................................................... 65 Capsaicin synergizes with