Chen, Shuyan. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2008. / Includes bibliographical references (leaves 91-98). / Abstracts in English and Chinese. / ABSTRACT --- p.i / ACKNOWLEGGMENTS --- p.v / LIST OF FIGURES --- p.vi / LIST OF TABLES --- p.vii / ABBREVIATIONS --- p.viii / CONTENTS --- p.ix / Chapter Chapter I --- Introduction / Chapter 1.1 --- Introduction of BRE / Chapter 1.1.1 --- Discovery of BRE --- p.1 / Chapter 1.1.2 --- Isoforms of BRE --- p.2 / Chapter 1.1.3 --- Homology and orthologs of BRE --- p.3 / Chapter 1.1.4 --- Expression studies of BRE mRNA --- p.4 / Chapter 1.1.5 --- Expression and cellular localization of BRE protein --- p.5 / Chapter 1.1.6 --- Interaction between BRE and death receptor --- p.6 / Chapter 1.1.7 --- Anti-apoptotic effect of BRE in cell line studies --- p.9 / Chapter 1.1.8 --- Anti-apoptotic effect of BRE in vivo --- p.11 / Chapter 1.1.9 --- BRE's role in DNA repair and ubiquitination --- p.12 / Chapter 1.1.10 --- BRE's role in regulation of Prohibitin and p53 expression --- p.13 / Chapter 1.2 --- Hepatocellular carcinoma / Chapter 1.2.1 --- Carcinogenesis --- p.15 / Chapter 1.2.2 --- Diethylnitrosamine -induced HCC --- p.15 / Chapter 1.2.3 --- Mouse model for HCC studies --- p.17 / Chapter 1.2.4 --- BRE in human HCC --- p.18 / Chapter 1.3 --- Green Fluorescent Protein / Chapter 1.3.1 --- Application of GFP in biological research --- p.19 / Chapter 1.3.2 --- Advantage of GFP applied in protein localization --- p.19 / Chapter Chapter II --- Materials and Methods / Chapter 2.1 --- Materials / Chapter 2.1.1 --- Primer used for cloning --- p.20 / Chapter 2.1.2 --- DNA clones used in the studies --- p.21 / Chapter 2.1.3 --- Materials for DNA manipulation --- p.24 / Chapter 2.1.4 --- Materials for protein manipulation --- p.24 / Chapter 2.1.5 --- Antibodies --- p.25 / Chapter 2.1.6 --- Chemicals --- p.25 / Chapter 2.1.7 --- Kits --- p.26 / Chapter 2.1.8 --- Culture media and reagents --- p.26 / Chapter 2.1.9 --- Bacterial strain used for transformation and cloning --- p.26 / Chapter 2.1.10 --- Instrumentation --- p.27 / Chapter 2.1.11 --- Animals --- p.27 / Chapter 2.1.12 --- Slides --- p.27 / Chapter 2.2 --- Methods / Chapter 2.2.1 --- Construction of Plasmids / Chapter 2.2.1.1 --- Polymerase chain reaction (PCR) --- p.28 / Chapter 2.2.1.2 --- Enzyme Digestion and Ligation --- p.29 / Chapter 2.2.1.3 --- Transformaion / Chapter 2.2.1.3.1 --- Preparation of competent cells --- p.29 / Chapter 2.2.1.3.2 --- Heat-shock Transformation --- p.29 / Chapter 2.2.1.4 --- Midi Prep of plasmids --- p.30 / Chapter 2.2.2 --- Cell Culture --- p.30 / Chapter 2.2.3 --- Transfection --- p.30 / Chapter 2.2.4 --- MG-132 treatment --- p.31 / Chapter 2.2.5 --- Flow Cytometry --- p.32 / Chapter 2.2.6 --- Western blotting / Chapter 2.2.6.1 --- SDS-PAGE --- p.32 / Chapter 2.2.6.2 --- Immunoblotting --- p.32 / Chapter 2.2.7 --- Production of Monoclonal Antibody --- p.33 / Chapter 2.2.8 --- Mice --- p.34 / Chapter 2.2.9 --- Tissue Processing --- p.35 / Chapter 2.2.10 --- Tissue Section --- p.35 / Chapter 2.2.11 --- Immunostaining --- p.36 / Chapter 2.2.12 --- H&E staining --- p.36 / Chapter 2.2.13 --- Picture Capture --- p.37 / Chapter 2.2.14 --- Confocal imaging --- p.37 / Chapter 2.2.14 --- Statistical Analysis --- p.37 / Chapter Chapter III --- BRE promotes growth of chemically-induced hepatocellular carcinoma / Chapter 3.1 --- DEN induced HCC in male mice --- p.38 / Chapter 3.2 --- BRE facilitates HCC in female mice --- p.44 / Chapter 3.3 --- Over-expression of BRE in tumor portion --- p.45 / Chapter 3.4 --- Direct effect of DEN on BRE expression --- p.47 / Chapter 3.5 --- Contribution of infiltrating cells in up-regulation of BRE --- p.50 / Chapter Chaper IV --- Subcellular localization of BRE / Chapter 4.1 --- GFP-BRE fusion constructs --- p.55 / Chapter 4.1.1 --- Transfection of GFP-BRE fusions --- p.58 / Chapter 4.1.2 --- Flow cytometry analysis of GFP-BRE fusions --- p.59 / Chapter 4.1.3 --- Western blot analysis of GFP-BRE fusions --- p.62 / Chapter 4.1.4 --- Stabilities of GFP-BRE fusions --- p.64 / Chapter 4.2 --- Fusions between GFP and the deletion mutants of BRE --- p.66 / Chapter 4.2.1 --- Transfection of mutants --- p.68 / Chapter 4.2.2 --- Low expression of mutants --- p.69 / Chapter 4.3 --- MG-132 treatments / Chapter 4.3.1 --- Increased expression of fusion proteins --- p.74 / Chapter 4.3.2 --- Subcellular localization of GFP-BRE fusions --- p.77 / Chapter Chapter V --- Discussion / Chapter 5.1 --- Functional role of BRE in HCC / Chapter 5.1.1 --- Stage model of carcinogenesis --- p.81 / Chapter 5.1.2 --- Anti-apoptotic genes in cancer --- p.84 / Chapter 5.1.3 --- Limitation of the study --- p.85 / Chapter 5.1.4 --- Conclusion --- p.85 / Chapter 5.2 --- Subcellular localization of BRE / Chapter 5.2.1 --- Low expression of GFP-BRE fusions --- p.86 / Chapter 5.2.2 --- Additional study --- p.90 / Chapter 5.2.3 --- Conclusion --- p.90 / Reference --- p.91 / Appendix --- p.99
| Identifer | oai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_326463 |
| Date | January 2008 |
| Contributors | Chen, Shuyan., Chinese University of Hong Kong Graduate School. Division of Chemical Pathology. |
| Source Sets | The Chinese University of Hong Kong |
| Language | English, Chinese |
| Detected Language | English |
| Type | Text, bibliography |
| Format | print, xii, 109 leaves : ill. (some col.) ; 30 cm. |
| Rights | Use of this resource is governed by the terms and conditions of the Creative Commons “Attribution-NonCommercial-NoDerivatives 4.0 International” License (http://creativecommons.org/licenses/by-nc-nd/4.0/) |
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