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The Function of Δ40p53 in Breast Cancers, We Utilised Gene Overexpression As Well As Rnai Techniques to Establish Breast Cancer Cell Line Models

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The Function of Δ40p53 in Breast Cancers, We Utilised Gene Overexpression As Well As Rnai Techniques to Establish Breast Cancer Cell Line Models The Function of Δ40p53 in Breast Cancer Xiajie Zhang M. Sc. (Dresden, Germany) B. Sc. (Beijing China) Doctor of Philosophy (Medical Genetics) University of Newcastle Submitted for examination November 2019 Declarations STATEMENT OF ORIGINALITY I hereby certify that the work embodied in the thesis is my own work, conducted under normal supervision. The thesis contains no material which has been accepted, or is being examined, for the award of any other degree or diploma in any university or other tertiary institution and, to the best of my knowledge and belief, contains no material previously published or written by another person, except where due reference has been made. I give consent to the final version of my thesis being made available worldwide when deposited in the University’s Digital Repository, subject to the provisions of the Copyright Act 1968 and any approved embargo. Xiajie Zhang 15/11/2019 STATEMENT OF COLLABORATION I hereby certify that the work embodied in this thesis has been done in collaboration with other researchers. I have included below a statement outlining the extent of collaboration, with whom and under what auspices. CONTRIBUTION BY OTHERS TO THE THESIS There were a number of collaborators who have made important contributions to the unpublished work presented in this thesis as follows: A • Dr Hamish Campbell and Professor Antony Braithwaite generated the MCF-7-LeGO and MCF- 7Δ40p53 cell lines. • Dr Brianna Catherine Morten, being my co-supervisor and lab colleague, helped the work throughout investigation of cell line responses to DNA-damage. B Acknowledgement This thesis would not have been completed without the help and support of a number of wonderful people. I truly and will forever appreciate the guidance and friendship throughout this PhD journey and would like to acknowledge them for making this milestone possible. Firstly, I would like to thank my supervisors, Professor Rodney Scott and Kelly Avery-Kiejda, for giving me this chance to come to Australia. Rodney, thank you for your prompt reply to my inquiry and this amazing opportunity to work in the awesome genomic cancer research facility. Kelly, I cannot thank you more for all the guidance and help throughout these four years and the encouraging push for me to be brave and independent. I cannot forget the conference journeys with you, when you were certainly not just a supervisor, but more like a friend I trust and open up to. A special thank-you to my co-supervisor Brianna Morten. You are such a cheerful soul, and best work mate one can ask for, I will always remember our nerdy chat over Δ40p53, thank you for sharing the knowledge with me. Another special thank-you to my co-supervisor Rick Thorne, the inspiration and guidance you have offered are amazingly instructive. Thank you for making yourself always available for me. To others who have kindly offered help in my data analysis, Carlos, Heather and Sean, thank you for being patient listeners and teachers. Sean, thank you very much as my swimming instructor and also for being such a bright spirit. To the team of breast cancer research group, present and past, thank you for your help and support in life and at work. Andrea, you were my one and only connection here to my life back in Dresden and I could not forget your generous help. Katherine, Mamta, Maddi, Luiza and Coralie, the friendship with you girls is a treasure for me. To all friendly faces at level 3 west, Ellise, Kira, Samara, Kumar, Dani, Michelle and many many more, thank you for being such understanding and kind friends; Ann, Megan and many many more, thanks for the sympathy and help when I broke my wrist. Your kind words and support are truly a comfort. To my friends out of the lab, Debbie, you are one most stable existence here in Newcastle, I will never ever forget all the time with you, crazy, adventurous and full of joy. Thank you, Shankar, Candy, Aun, Yudi, Kelly and many more, the friendship with you is so important. To my family, Mum and Dad, without you, none of this would be possible. Thank you for letting your little girl to chase her dream, thank you for being respectful and supportive for my whole life, thank you for simply being there where I call a home! To my partner Alex, I feel so blessed and lucky to have you as my best friend. We almost go through this whole PhD together and get to know each other a bit more every day and yet I never have felt bored. I really appreciate your accompany, kindness, humour and positivity. I have become stronger C because of you. Thank you very much also for numerous discussion on experiments, bioinformatics and all sorts of scientific topics. Finally, I would like to acknowledge the University of Newcastle for this fantastic opportunity to do a PhD and for providing me the UNIPRS and UNRSC5050 scholarships. A further acknowledgement goes to the Hunter Medical Research Institute for letting me conduct my research in such fabulous research facility, and to Hunter Cancer Research Alliance for providing incredible opportunities in research support, communication and personal development. D This thesis is dedicated to the most loving, caring and supportive Mom and Dad, and to my wonderful partner Alex. Without your generous devotion, I could not have accomplished this milestone. Thank you. E List of publications and oral conference presentations Publication: Zhang X, Morten BC, Scott RJ, Avery-Kiejda KA. A Simple Migration/Invasion Workflow Using an Automated Live-cell Imager. JoVE. 2019(144):e59042. Oral presentations: Zhang X, Möller FJ, Vollmer G: “Impact of a prenatally started and continuously maintained soy-derived isoflavones (sISO) exposure on mammary glands of ovary-intact ACI rats”, The 2015 Hunter Cancer Research Symposium, rapid poster session, Newcastle, Australia, 27th November 2016. - Zhang X, Morten BC, Campbell H, Braithwaite A, Scott RJ, Avery-Kiejda KA: “Δ40p53 regulates migration in breast cancer cell line MCF-7 cells carrying wild type p53”, The 2016 Hunter Cancer Research Symposium, Newcastle, Australia, 25th November 2016. Zhang X, Morten BC, Campbell H, Braithwaite A, Scott RJ, Avery-Kiejda KA: “Δ40p53 has a potential role in regulating EMT”, HCRA, ECR and PhD Students Seminar, Newcastle, Australia, 29th March, 2017. Zhang X: “Little Bro, Big Role”, 3MT Competition, Newcastle, Australia, 31st May 2017. Zhang X, Morten BC, Campbell H, Braithwaite A, Scott RJ, Avery-Kiejda KA: “Δ40p53 has a potential role in regulating EMT in MCF-7 breast cancer cells”, Australian Metastasis Research Society Annual Scientific Meeting, Sydney, Australia, 15th November 2017. - Zhang X, Morten BC, Campbell H, Braithwaite A, Scott RJ, Avery-Kiejda KA: “The role of Δ40p53 in EMT in breast cancer”, ASMR Hunter Region Satellite Scientific Meeting, Newcastle, Australia, 1st June 2018. Zhang X, Morten BC, Campbell H, Braithwaite A, Avery-Kiejda KA: “Characterisation of transcriptional targets regulated by Δ40p53 in response to DNA damage in breast cancer cells”, 4th International p53 isoform workshop, flash poster session, Dubrovnik, Croatia, 3-6th November, 2019. F Table of Contents DECLARATIONS ............................................................................................................................................... A ACKNOWLEDGEMENT ..................................................................................................................................... C LIST OF PUBLICATIONS AND ORAL CONFERENCE PRESENTATIONS .................................................................. F TABLE OF CONTENTS ........................................................................................................................................ I ABSTRACT ......................................................................................................................................................... I LIST OF FIGURES ..............................................................................................................................................III LIST OF TABLES ............................................................................................................................................... V LIST OF ABBREVIATIONS ................................................................................................................................ VI INTRODUCTION .................................................................................................................. - 2 - 1.1 OVERVIEW ............................................................................................................................................ - 2 - 1.2 BREAST CANCER ..................................................................................................................................... - 3 - 1.2.1 Breast cancer and its risk factors .................................................................................................. - 4 - 1.2.2 Hormones and their receptors ...................................................................................................... - 5 - 1.2.3 Classification of breast cancer ...................................................................................................... - 6 - 1.2.4 Breast cancer initiation and metastasis ........................................................................................ - 7 - 1.3 THE P53 TUMOUR SUPPRESSOR PROTEIN .................................................................................................. - 10 - 1.3.1 The p53 pathway .......................................................................................................................
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