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Acombination of Chinese Herb Hedyotis Diffusa and Scutellaria

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Acombination of Chinese Herb Hedyotis Diffusa and Scutellaria A combination of Chinese herb Hedyotis diffusa and Scutellaria barbata impairs mitotic entry of prostate cancer cells without eliciting DNA damage By Su Su Thae Hnit A thesis submitted in satisfaction of the requirements for the degree of Doctor of Philosophy School of Science and Health Western Sydney University Oct 2017 Primary Supervisor: Associate Professor Qihan Dong Co-supervisors: Professor Paul De Souza, Professor Alan Bensoussan, Dr Mu Yao 1 Statement of Authentication This thesis is submitted in fulfillment of the requirements of the degree of Doctor of Philosophy in the School of Science and Health at Western Sydney University. The work presented in this thesis is, to the best of my knowledge, original except as acknowledged in the text. I hereby declare that I have not previously submitted this material, either in whole or in part, for a degree or a diploma in any other institution. Su Su Thae Hnit Oct 2017 2 Acknowledgments This thesis would not have been possible without the help of a number of people. These people offered me so much that I could ever imagine and made me who I am now. First and foremost, my deepest gratitude goes to my primary supervisor, Associate Professor Qihan Dong. I would like to say thank you very much from the bottom of my heart for generously lavishing me with abundant guidance, mentorship, knowledge, expert advice, attention, care and support. I will never forget your advice on fundamental thinking and paying attention in details. I can’t even express how much I appreciate for all of your help. I greatly appreciate all the time and patience you have dedicated to take me on science world. It has been a privilege to be part of your research team and really grateful for the useful comments, remarks and engagement through the learning process of this thesis. Also, your constant mentoring and encouragement is highly valued not only in this project but also for my future carrier. You are absolutely respectable and admirable supervisor. My special thanks go to Dr Mu Yao for his expert advice on experimental support, constructive criticism and encouragement throughout the years. Your help has been enormous in the process of my PhD, especially in the hard time of problem solving, and experimental method improvement. Your advice and support in various immunolabelling techniques have been priceless. You enlightened my understanding of science into whole new level. I would also like to thank to Professor Paul De Souza and Professor Alan Bensoussan for advice in thesis structure, assistance in proofreading and examining my thesis. Thank you very much Professor Li Zhong for providing DR recipe and Dr Cheng Khoo for chemical 3 profiling of DR. I also thank Associate Professor Natalka Suchowerska for providing the facility of radiation treatment for my experiments. I also thank my lab fellows Dr Amber Xie, fellow PhD student Ling Bi and Honours student Sally Hanna Rie for their great assistance and advice on my work. Thanks must also go to Honours student William Wei for providing the RT-PCR data and his enthusiastic questions. I must mention ‘Ladies who lunch together’ for their encouragement throughout my PhD experience. To those in the School of Science and Health, University of Western Sydney and Charles Perkin Centre who have assisted me in my PhD journey, I am truly appreciated for their contributions. Last not the least, I would like to thank my Dad and Mum for their love and support in helping me to get through my PhD years. My words can’t express how grateful I am and it would have been absolutely impossible for me to get this far if it wasn’t for the support and encouragement from my family. 4 Table of Contents Statement of Authentication .................................................................................................... 2 Acknowledgments ..................................................................................................................... 3 Table of Contents ..................................................................................................................... 5 List of Figures ......................................................................................................................... 11 List of Tables ........................................................................................................................... 14 List of abbreviations ............................................................................................................... 15 Publication .............................................................................................................................. 19 Conference Abstracts ............................................................................................................. 19 List of Presentations ............................................................................................................... 20 Abstract ................................................................................................................................... 21 1 Chapter 1 - Introduction .......................................................................................... 25 1.1 Prevalence of PCa ..................................................................................................... 26 1.2 Treatment options ..................................................................................................... 27 1.2.1 Surgery .................................................................................................................. 27 1.2.2 Radiation therapy .................................................................................................. 27 1.2.3 Androgen Deprivation Therapy (ADT) ................................................................ 28 1.2.4 Chemotherapy ....................................................................................................... 29 1.2.5 Active Surveillance (AS) ...................................................................................... 29 1.3 Cell Cycle and Key regulators ................................................................................. 31 1.3.1 The importance of Cyclins and CDKs in cell cycle ............................................. 31 1.3.2 Cell cycle checkpoints .......................................................................................... 34 1.3.2.1 G1-S checkpoint .............................................................................................. 35 1.3.2.2 G2-M checkpoint ............................................................................................. 36 1.3.2.3 Mitosis checkpoint or spindle assembly checkpoint (SAC) ........................... 38 1.4 Signalling pathways and proteins involved in DNA damage response ................ 41 1.4.1 Activation of ATM and ATR ............................................................................... 42 1.4.2 Activation of CHK2 and CHK1 ........................................................................... 44 5 1.4.3 ATM-CHK2, ATR-CHK1 Signal transduction .................................................... 45 1.4.4 Aberrant regulation of ATM and ATR ................................................................. 50 1.4.5 Aberrant regulation of CHK2 and CHK1 ............................................................. 50 1.4.6 γH2AX................................................................................................................... 51 1.4.7 53BP1 ................................................................................................................... 54 1.5 M phase promoting complex and regulators at G2 to M phase transition ........... 56 1.5.1 Cyclin B1-CDK1 complex ................................................................................... 56 1.5.1.1 Role of Cyclin B1 and CDK1 at G2/M phase ................................................. 56 1.5.1.2 Regulation of Cyclin B1-CDK1 ..................................................................... 57 1.5.1.2.1 Regulation by Cyclin B1 .......................................................................... 57 1.5.1.2.2 Regulation by CAK .................................................................................. 58 1.5.1.2.3 Regulation by Myt1 and Wee1 ................................................................. 58 1.5.1.2.4 Regulation by CDC25C ........................................................................... 59 1.5.1.2.5 Regulation by intracellular localization ................................................... 60 1.5.2 PLK1 ..................................................................................................................... 62 1.5.2.1 Role of PLK1 in G2-M transition .................................................................... 63 1.5.2.2 Role of PLK1 in DNA damage response ........................................................ 64 1.5.2.3 Role of PLK1 in Mitosis ................................................................................. 66 1.5.3 Aurora A ............................................................................................................... 68 1.5.3.1 Role of Aurora A in G2-M transition .............................................................. 68 1.5.3.2 Role of Aurora A in DNA damage response .................................................. 69 1.5.3.3 Role of Aurora A in Mitosis ........................................................................... 70 1.6 The use of Traditional Chinese Medicine (TCM)
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