Mak, Keng Hou. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2011. / Includes bibliographical references (leaves 77-91). / Abstracts in English and Chinese. / Thesis Committee --- p.2 / Declaration --- p.3 / Table of Contents --- p.4 / List of Abbreviations --- p.6 / List of Figures --- p.8 / Abstract --- p.10 / Chapter Chapter 1 --- Introduction --- p.12 / Chapter 1.1 --- Background --- p.12 / Chapter 1.1.1 --- Overview of apoptosis --- p.12 / Chapter 1.1.2 --- Synopsis of the apoptotic pathway --- p.13 / Chapter 1.1.3 --- Defining apoptosis --- p.14 / Chapter 1.1.4 --- Interaction between pro- and anti-apoptotic factors determines cell fate --- p.14 / Chapter 1.1.5 --- DNA fragmentation during the execution phase --- p.15 / Chapter 1.1.6 --- Current understanding of the point of commitment in apoptosis --- p.16 / Chapter 1.1.7 --- Previous studies and hypotheses related to the reversibility of late-state apoptosis --- p.16 / Chapter 1.1.8 --- Unanswered questions --- p.19 / Chapter 1.2 --- "Hypothesis and objectives, Study models and Significance" --- p.19 / Chapter 1.2.1 --- Hypothesis and objectives --- p.19 / Chapter 1.2.2 --- Study models --- p.20 / Chapter 1.2.3 --- Significance --- p.20 / Chapter Chapter 2 --- Materials and Methods --- p.22 / Chapter Chapter 3 --- Results --- p.30 / Chapter 3.1 --- Dying cells reversed execution stage of apoptosis after removal of apoptotic stimuli --- p.30 / Chapter 3.2 --- Dying cells reversed apoptosis after DNA damage --- p.37 / Chapter 3.3 --- Genetic alterations and transformation occurred after reversal of apoptosis --- p.43 / Chapter 3.4 --- Investigating molecular mechanism driving reversal of apoptosis --- p.50 / Chapter 3.4.1 --- Preparation and characterization of samples for microarray --- p.50 / Chapter 3.4.2 --- Gene ontology enrichment analysis of the expression profile during reversal of apoptosis --- p.52 / Chapter 3.4.3 --- Interfering stress response or anti-apoptotic factors during the reversal of apoptosis drove cells to terminal death --- p.56 / Chapter Chapter 4 --- Discussion --- p.62 / Chapter 4.1 --- "Reversal of apoptosis in ""normal cells"" was observed" --- p.62 / Chapter 4.2 --- Cells surviving apoptosis had their genomes damaged and altered --- p.63 / Chapter 4.3 --- Transformation occurred after reversal of apoptosis --- p.65 / Chapter 4.4 --- Investigating molecular mechanism driving reversal of apoptosis --- p.65 / Chapter 4.5 --- Summary --- p.68 / Chapter Chapter 5 --- Perspectives --- p.70 / Chapter 5.1 --- Could reversal of apoptosis be evolutionarily advantageous? --- p.70 / Chapter 5.2 --- "Reversal of apoptosis as an ""individualistic"" behavior against organismal integrity" --- p.71 / Chapter 5.3 --- Proposed studies --- p.72 / Chapter 5.3.1 --- Other apoptotic targets that may leave persistent effects --- p.72 / Chapter 5.3.2 --- Post- caspase activation regulation of apoptosis --- p.74 / Chapter 5.3.3 --- Identifying correlation between reversal of apoptosis and cancer --- p.74 / Chapter 5.3.4 --- Single cell methods and cell tracking system for further studies --- p.75 / Chapter 5.3.5 --- Notes on studying reversal of apoptosis in relation to phagocytosis --- p.76 / References --- p.77

Wong, Wan Man. / Thesis submitted in: December 2006. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2007. / Includes bibliographical references (leaves 85-98). / Abstracts in English and Chinese. / Abstract --- p.i / Abstract in Chinese --- p.iii / Acknowledgements --- p.v / Lists of Figures and Tables --- p.vi / Table of Abbreviations --- p.xi / Table of Contents --- p.xv / Chapter Chapter I --- Introduction / Chapter 1.1 --- Brain and Reproductive Organ Expressed Gene --- p.1 / Chapter 1.2 --- Programmed cell death --- p.4 / Chapter 1.3 --- Limb development in mouse --- p.8 / Chapter 1.4 --- Role of BRE in apoptosis --- p.12 / Chapter 1.5 --- Role of programmed cell death in interdigital tissue regression --- p.14 / Chapter 1.6 --- Aim of study --- p.17 / Chapter Chpater II --- Materials and methods / Chapter 2.1 --- Mice --- p.18 / Chapter 2.2 --- In-situ hybridization / Chapter 2.2.1 --- Histology --- p.18 / Chapter 2.2.2 --- Preparation of riboprobe for in-situ hybridization --- p.19 / Chapter 2.2.3 --- In-situ hybridization --- p.20 / Chapter 2.3 --- Interdigital tissue culture --- p.21 / Chapter 2.4 --- Gene interference / Chapter 2.4.1 --- Construction of Bre-siRNA --- p.22 / Chapter 2.4.2 --- siRNA transfection of cultured interdigital cells --- p.23 / Chapter 2.5 --- Semi-quantitative RT-PCR / Chapter 2.5.1 --- Sample collection of interdigital cells and explants --- p.23 / Chapter 2.5.2 --- RNA isolation and extraction --- p.24 / Chapter 2.5.3 --- Reverse-transcription and cDNA synthesis --- p.25 / Chapter 2.5.4 --- Polymerase chain reaction --- p.26 / Chapter 2.6 --- Assay of cell viability by MTT --- p.28 / Chapter 2.7 --- Comparative proteomics --- p.30 / Chapter 2.7.1 --- Collection of interdigital cells --- p.30 / Chapter 2.7.2 --- Preparation of cell lysate --- p.31 / Chapter 2.7.3 --- Assay of protein concentration in cell lysate --- p.31 / Chapter 2.7.4 --- Two-dimensional gel electrophoresis --- p.33 / Chapter 2.7.5 --- Protein identification by mass fingerprinting --- p.36 / Chapter 2.8 --- Statistical Method --- p.38 / Chapter Chapter III --- Results / Chapter 3.1 --- Spatial and temporal expression of Bre in murine embryonic hindlimbs --- p.39 / Chapter 3.2 --- Expression of Bre isoforms in interdigital tissues --- p.45 / Chapter 3.3 --- Silencing of Bre expression by siRNA in interdigital cells --- p.49 / Chapter 3.4 --- Effect on viability of Bre-silenced interdigital cells by siRNA --- p.51 / Chapter 3.5 --- Comparative proteomic profile of Bre-silenced interdigital cultured cells --- p.53 / Chapter 3.6 --- Identification of proteins that were differentially expressed by MALDI- TOF --- p.71 / Chapter 3.7 --- The mRNA levels of proteins identified that were differentially expressed --- p.74 / Chapter Chapter IV --- Discussion --- p.77 / References --- p.85 / Appendices --- p.99 / Publication --- p.108

Li Qing. / "July 2004." / Thesis (Ph.D.)--Chinese University of Hong Kong, 2004. / Includes bibliographical references (p. 161-192). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Mode of access: World Wide Web. / Abstracts in English and Chinese.

Breast cancer, as the most prevalent cancer in women, is responsible for more than 15% of new cancer cases and about 6.9% of all cancer-related death in the US. A major cause of therapeutic failure in breast cancer is the development of resistance to chemotherapy, especially for triple-negative breast cancer (TNBC). Therefore, how to overcome chemoresistance is the major challenge to improve the life expectancy of breast cancer patients. Our studies demonstrate that TNBC cells surviving the chronic treatment of chemotherapeutic drugs show significantly higher expression of the dual serine/threonine and tyrosine protein kinase (DSTYK) than non-treated parental cells. In our in vitro cellular models, DSTYK knockout via the CRISPR/Cas9-mediated technique results in apoptotic cell death of chemoresistant cells upon drug treatment. Moreover, DSTYK knockout promotes chemotherapeutic drug-induced tumor cell death in an orthotopic mouse model. These findings suggest that DSTYK exerts an important and previously unknown role in promoting chemoresistance. Our studies provide fundamental insight into the role of DSTYK in chemoresistance in TNBC cells and lay the foundation for the development of new strategies targeting DSTYK for improving TNBC therapy.

Fibroblast growth factor receptors (FGFR) play key roles in proliferation, differentiation, and tumorigenesis. Many urothelial carcinomas contain activating point mutations or increased expression of FGFR3. However, little is known about the role of other FGFRs. We examined FGFR expression in telomerase-immortalized normal human urothelial cells, urothelial carcinoma cell lines, and tumor samples and showed that FGFR1 expression is increased in a high proportion of cell lines and tumors independent of stage and grade. To determine the role of FGFR1 in low-stage bladder cancer, we overexpressed FGFR1 in telomerase-immortalized normal human urothelial cells and examined changes in proliferation and cell survival in response to FGF2. FGFR1 stimulation increased proliferation and reduced apoptosis. To elucidate the mechanistic basis for these alterations, we examined the signaling cascades activated by FGFR1. FRS2alpha and PLCgamma were activated in response to FGF2, leading to activation of the mitogen-activated protein kinase pathway. The level of mitogen-activated protein kinase activation correlated with the level of cyclin D1, MCL1, and phospho-BAD, which also correlated with FGFR-induced proliferation and survival. Knockdown of FGFR1 in urothelial carcinoma cell lines revealed differential FGFR1 dependence. JMSU1 cells were dependent on FGFR1 expression for survival but three other cell lines were not. Two cell lines (JMSU1 and UMUC3) were dependent on FGFR1 for growth in soft agar. Only one of the cell lines tested (UMUC3) was frankly tumorigenic; here, FGFR1 knockdown inhibited tumor growth. Our results indicate that FGFR1 has significant effects on urothelial cell phenotype and may represent a useful therapeutic target in some cases of urothelial carcinoma.