The role of DOK7 and its variants in colorectal cancer By Lucy Kate Satherley Cardiff-China Medical Research Collaborative Cardiff University School of Medicine December 2016 Thesis submitted to Cardiff University for the application of degree of Doctor of Philosophy (PhD) The role of DOK7 and its variants in colorectal cancer Student: Lucy Kate Satherley Student Number: 990334242 Supervisors: Professor WG Jiang Miss R Hargest Doctor L Ye Cardiff-China Medical Research Collaborative Cardiff University School of Medicine Heath Park Cardiff Wales UK CF14 4XN December 2016 II Declaration This work has not been submitted in substance for any other degree or award at this or any other university or place of learning, nor is being submitted concurrently in candidature for any degree or other award. Signed……………………………………Date………… Statement 1 This thesis is being submitted in partial fulfilment of the requirements for the degree of PhD Signed……………………………………Date……… Statement 2 This thesis is the result of my own independent work/investigation, except where otherwise stated. Other sources are acknowledged by explicit references. The views expressed are my own. Signed……………………………………Date………… III Statement 3 I hereby give consent for my thesis, if accepted, to be available for photocopying and for interlibrary loan, and for the title and summary to be made available to outside organisations. Signed……………………………………Date………… IV Dedication To my wonderful parents For inspiring in me a love of learning and for your unfailing love and support, this thesis is dedicated to you with all my love and thanks for all you have done for me. V Acknowledgements I would like to thank my supervisors, Professor Wen G. Jiang and Miss Rachel Hargest, firstly, for providing me with the opportunity to participate in laboratory based research and secondly, for their guidance and support over the last four years. I would also like to thank all the staff and students at the Cardiff-China Medical Research Collaborative for making me feel so welcome. It has been such a privilege to combine clinical medicine and science and your help, advice, support and friendship has made my time as a PhD student so enjoyable. In particular, I would like to thank my laboratory supervisor Dr. Lin Ye for his friendship, support, encouragement and advice. I am also very grateful to my clinical colleagues, particularly those in the Department of Surgery at the University Hospital of Wales in Cardiff, who have allowed me to dedicate time to my research and also to Cardiff and Vale NHS Trust for employing me as a clinical fellow during my PhD studies. I would also like to especially thank my consultant colleagues at the Breast Centre in University Hospital Llandough (Ms Eleri Davies, Ms Donna Egbeare, Mr Sumit Goyal and Professor Helen Sweetland) whose support and encouragement has ensured I have been able to write up, submit, defend and complete my thesis whilst continuing my higher surgical training. Finally, I would like to dedicate this work to my family, especially my parents, and to my husband Guy, and to thank them for their never-ending love, support and encouragement through all the challenges of working full-time and simultaneously undertaking a full-time PhD! VI Publications and Presentations undertaken during the period of research Published papers LK Satherley, PH Sun, K Ji, M Mason, R Hargest, WG Jiang, L Ye. Prostate apoptosis response-4 (PAR4) suppresses growth and invasion of breast cancer cells and is positively associated with patient survival. Anticancer Research 2016 36:1227-1236 (Impact factor = 1.895). MFA Malik, LK Satherley, EL Davies, L Ye, WG Jiang. Expression of Semaphorin 3C in breast cancer and its impact on adhesion and invasion of breast cancer cells. Anticancer Research 2016 36:1281-1286 (Impact factor = 1.895). S Shao, PH Sun, LK Satherley, X Gao, K Ji, Y Feng, Y Jia, J Ji, WG Jiang, L Ye. Reduced RanBPM expression is associated with distant metastasis in gastric cancer and chemoresistance. Anticancer Research 2016 36: 1295-1304 (Impact factor = 1.895). J Li, L Ye, PH Sun, LK Satherley, R Hargest, Z Zhang, WG Jiang. MTA1 is up- regulated in colorectal cancer and is inversely correlated with lymphatic metastasis. Cancer Genomics and Proteomics 2015 11-12;12(6):339-345 (Impact factor = 2.800). LK Satherley, L Ye, WG Jiang, R Hargest. Tumour suppressor genes in gastric carcinogenesis: An up-to-date review of genetic and epigenetic changes in the development of gastric adenocarcinoma. Transl Gastrointest Cancer 2015;4(4):294-307. VII WG Jiang, L Ye, F Ruge, PH Sun, AJ Sanders, K Ji, J Lane, L Zhang, L Satherley, HP Weeks, X Zhi, Y Gao, C Wei, Y Wu, MD Mason. Expression of Sonic Hedgehog (SHH) in human lung cancer and the impact of YangZheng XiaoJi on SHH-mediated biological function of lung cancer cells and tumour growth. Anticancer Research 2015 Mar;35(3):1321-31 (Impact factor = 1.895). WG Jiang, L Ye, F Ruge, S Owen, T Martin, PH Sun, AJ Sanders, J Lane, L Satherley, HP Weeks, Y Gao, C Wei, Y Wu, MD Mason. YangZheng XiaoJi exerts anti-tumour growth effects by antagonising the effects of HGF and its receptor, cMET in human lung cancer cells. J Transl Med 2015 Aug 27;13:280 (Impact factor = 3.694). Published abstracts LK Satherley, L Ye, R Hargest, WG Jiang. DOK7 variant expression and function in human colorectal cancer. Anticancer Research 2015 35 (7): 4331 (Impact factor = 1.895). WG Jiang, L Ye, F Ruge, PH Sun, AJ Sanders, K Ji, J Lane, L Satherley, HP Weeks, Y Gao, C Wei, Y Wu, MD Mason. YangZheng Xiaoji sensitises the response to SIS3 inhibitor in human cancer cells, in vitro and in vivo; therapeutic implications. Anticancer Research 2015 35(7): 4360 (Impact factor = 1.895) L Satherley, W Jiang, R Hargest, J Torkington, M Davies. Expression of lymphangiogenic factors VEGF-C and VEGF-D and impact of VEGF-D knockout on tubule formation in human colorectal cancer. Br J Surg 2015 102: 8 (Impact factor = 5.596). VIII LK Satherley, L Ye, R Hargest, WG Jiang. DOK7 expression in colorectal cancer cells and association with clinical and prognostic outcome. Br J Surg 2014 101(Suppl 4): 12 (Impact factor = 5.596). L Satherley, L Ye, R Hargest, WG Jiang. DOK7 expression and association with patient survival. European Journal of Cancer 2013 49:S158 (Impact factor = 6.163) Oral Presentations L Satherley. Dok7 expression in colorectal cancer: Association with tumour pathology and clinical outcome, Institute of Cancer and Genetics PhD seminar Day, Cardiff University, October 2014. L Satherley, L Ye, R Hargest, WG Jiang. Dok7 expression in colorectal cancer cells and association with clinical and prognostic outcome. MIA Prize session, President’s Day meeting, Royal Society of Medicine, June 2014 L Satherley, WG Jiang, R Hargest, J Torkington, ML Davies. Expression of lymphangiogenic factors VEGF-C and VEGF-D and impact of VEGF-D knockout on tubule formation in human colorectal cancer. Association of Surgeons of Great Britain and Ireland (ASGBI) Conference (Short Paper of Distinction), Harrogate, May 2014 L Satherley, L Ye, R Hargest, W Jiang Dok7 expression in colorectal cancer cells and association with clinical and prognostic outcome, SARS conference, Cambridge, January 2014 IX L Satherley. Medical training in the UK. Cardiff University-Capital Medical University International Centre for Biomedical Research Symposium. Beijing, China, July 2013. Poster Presentations L Satherley, L Ye, R Hargest, WG Jiang. Downstream of tyrosine kinase 7 (DOK7) variant expression and function in human colorectal cancer. ECCO/ESMO Conference, Vienna, September 2015 C Bunston, PH Sun, L Satherley, K Ji, C Hao, J Ji, WG Jiang, L Ye. Capillary morphogenesis gene 2 is overexpressed by local advanced pancreatic tumours and the corresponding therapeutic potential. China-UK Cancer (CUKC) Conference, Cardiff, July 2015 L Satherley, L Ye, R Hargest, WG Jiang. DOK7 variant expression and function in human colorectal cancer. China-UK Cancer (CUKC) Conference, Cardiff, July 2015 WG Jiang, L Ye, F Ruge, PH Sun, AJ Sanders, K Ji, J Lane, L Satherley, HP Weeks, Y Gao, C Wei, Y Wu, MD Mason. YangZheng Xiaoji sensitises the response to SIS3 inhibitor in human cancer cells, in vitro and in vivo; therapeutic implications. China-UK Cancer (CUKC) Conference, Cardiff, July 2015 L Satherley, L Ye, R Hargest, WG Jiang. Dok 7 expression in colorectal cancer cells and association with clinical and prognostic outcome. BASO/Cardiff University teaching day, June 2014 X L Satherley, L Ye, R Hargest, WG Jiang. Psoriasin expression in human colorectal cancer. Digestive Diseases Week, Chicago, May 2014 L Satherley, L Ye, R Hargest, WG Jiang. DOK7 expression in colorectal cancer cells and association with patient survival. European Cancer Congress, Amsterdam, September 2013. XI Abbreviations α-SMA alpha smooth muscle actin Abl Abelson murine leukaemia viral oncogene ABS antibiotics AChE acetylcholine esterase AChR acetylcholine receptor ACPGBI Association of Coloproctology of Great Britain and Ireland AICR American Institute for Cancer Research AJCC American Joint Committee on Cancer Akt v-Akt murine thymoma viral oncogene (also known as PKB) Ang-1 angiopoietin-1 AP-1 activator protein 1 APC adenomatous polyposis coli APER abdomino-perineal excision of the rectum APS ammonium persulphate AFP alpha-fetoprotein ATP adenosine triphosphate B-ALL B-cell acute lymphoblastic leukaemia bcr breakpoint cluster region BCR B-cell antigen receptor BCR-ABL BCR-ABL fusion gene BM basement membrane bp base pairs BRCA Breast cancer gene