Lam Kin Bong Hubert. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2004. / Includes bibliographical references (leaves 139-161). / Abstracts in English and Chinese. / Abstract (in English) --- p.i / (in chinese) --- p.iv / Acknowledgements --- p.vi / Publications --- p.ix / Table of Contents --- p.x / List of Tables --- p.xiii / List of Figures --- p.xiv / List of Abbreviations --- p.xvi / Chapter CHAPTER1： --- General Introduction --- p.1 / Chapter Section 1.1 --- Historical Background and Application of Arsenic Trioxide as an Anti-cancer Agent --- p.1 / Chapter Section 1.2 --- Arsenic Trioxide Induces Apoptosis in Cancer Cells --- p.3 / Chapter 1.2.1 --- The Intrinsic and Extrinsic Pathways of Apoptosis Initiation --- p.4 / Chapter 1.2.2 --- The Convergence of Pathways --- p.8 / Chapter 1.2.3 --- Induction of Apoptosis by Arsenic Trioxide --- p.9 / Chapter 1.2.3.1 --- Controversies in the Involvement of the Extrinsic Pathway --- p.9 / Chapter 1.2.3.2 --- "Arsenic Trioxide, Oxidative Stress and the Mitochondria" --- p.10 / Chapter 1.2.3.3 --- Caspase-3 Activation in Arsenic Trioxide-mediated Apoptosis --- p.12 / Chapter Section 1.3 --- Arsenic Trioxide Perturbs the Cell Division Cycle --- p.13 / Chapter 1.3.1 --- The Cell Cycle Oscillator --- p.14 / Chapter 1.3.2 --- DNADamage and Cell Cycle Checkpoints --- p.15 / Chapter 1.3.3 --- Induction of Cell Cycle Arrest by Arsenic Trioxide and its Association with Apoptosis --- p.17 / Chapter Section 1.4 --- Acute Megakaryocytic Leukemia and Arsenic Trioxide --- p.20 / Chapter CHAPTER 2： --- Objectives --- p.28 / Chapter CHAPTER 3： --- Methodology --- p.30 / Chapter Section 3.1 --- Materials --- p.30 / Chapter Section 3.2 --- Methods --- p.39 / Chapter 3.2.1 --- Culture of Megakaryocytic Cells and Their Treatment with Arsenic Trioxide --- p.39 / Chapter 3.2.1.1 --- Maintenance of Cell Lines --- p.39 / Chapter 3.2.1.2 --- Treatment with Arsenic Trioxide --- p.39 / Chapter 3.2.2 --- "Effects of Arsenic Trioxide on Cell Proliferation, Apoptosis, Mitochondrial Integrity and Cell Division Cycle Profiles of Human Megakaryocytic Leukemia Cell Lines" --- p.40 / Chapter 3.2.2.1 --- Trypan Blue Exclusion Assay --- p.40 / Chapter 3.2.2.2 --- Quantitation of Externalized Phosphatidylserine --- p.41 / Chapter 3.2.2.3 --- Quantitation of Active Caspase-3 Expression --- p.42 / Chapter 3.2.2.4 --- Assessment of Mitochondrial Intensity --- p.42 / Chapter 3.2.2.5 --- Analysis of Cell Division Cycle Profile --- p.43 / Chapter 3.2.2.6 --- Analysis of Cell Cycle Kinetics by BrdU Labeling --- p.43 / Chapter 3.2.2.7 --- Identification of Cell Cycle Specificity of Caspase-3 Expression --- p.45 / Chapter 3.2.3 --- Effects of Arsenic Trioxide on the Expression of Apoptotic Signals in Human Megakaryocytic Leukemia Cell Lines --- p.45 / Chapter 3.2.3.1 --- Effects of Arsenic Trioxide on mRNA Expression Levels of Apoptotic Regulators --- p.45 / Chapter 3.2.3.2 --- Effects of Arsenic Trioxide on Protein Expression Levels of Apoptotic Regulators --- p.50 / Chapter 3.2.3.2.1 --- Flow Cytometric Analysis --- p.50 / Chapter 3.2.3.2.2 --- Western Blot Analysis --- p.51 / Chapter 3.2.4 --- Effects of Arsenic Trioxide on Gene Expression Profiles of Human Megakaryocytic Leukemia Cell Lines By Microarray Analysis --- p.54 / Chapter 3.2.5 --- Statistical Analysis --- p.57 / Chapter CHAPTER 4： --- "Effects of Arsenic Trioxide on Cell Proliferation, Apoptosis, Mitochondrial Integrity and Cell Division Cycle Profiles of Human Megakaryocytic Leukemia Cell Lines" --- p.62 / Chapter Section 4.1 --- Introduction --- p.62 / Chapter Section 4.2 --- Results --- p.63 / Chapter 4.2.1 --- Effects of Arsenic Trioxide on Proliferation Kinetics --- p.63 / Chapter 4.2.2 --- Effects of Arsenic Trioxide on Cell Viability --- p.64 / Chapter 4.2.3 --- Apoptosis-inducing Capability of Arsenic Trioxide --- p.65 / Chapter 4.2.3.1 --- Quantitation of Externalized Phosphatidylserine --- p.65 / Chapter 4.2.3.2 --- Quantitation of Active Caspase-3 Expression --- p.66 / Chapter 4.2.4 --- Effects of Arsenic Trioxide on Mitochondrial Integrity --- p.67 / Chapter 4.2.5 --- Effects of Arsenic Trioxide on Cell Division Cycle Profiles --- p.69 / Chapter 4.2.6 --- Effects of Arsenic Trioxide on Cell Cycle Kinetics by Bromodeoxyuridine Labeling --- p.69 / Chapter 4.2.7 --- Identification of Cell Cycle Specificity of Arsenic Trioxide-Induced Caspase-3 Activation --- p.71 / Chapter Section 4.3 --- Discussion --- p.72 / Chapter CHAPTER 5： --- Effects of Arsenic Trioxide on Apoptotic Signal Expression in Human Megakaryocytic Leukemia Cell Lines --- p.91 / Chapter Section 5.1 --- Introduction --- p.91 / Chapter Section 5.2 --- Results --- p.92 / Chapter 5.2.1 --- Effects of Arsenic Trioxide on mRNA Expression Levels of Apoptotic Regulators --- p.92 / Chapter 5.2.2 --- Effects of Arsenic Trioxide on Protein Expression Levels of Apoptotic Regulators --- p.94 / Chapter 5.2.2.1 --- Flow Cytometric Analysis --- p.94 / Chapter 5.2.2.2 --- Western Blot Analysis --- p.96 / Chapter Section 5.3 --- Discussion --- p.96 / Chapter CHAPTER 6： --- Effects of Arsenic Trioxide on Gene Expression Profiles of Human Megakaryocytic Leukemia Cell Lines by Microarray Analysis --- p.119 / Chapter Section 6.1 --- Introduction --- p.119 / Chapter Section 6.2 --- Results --- p.119 / Chapter Section 6.3 --- Discussion --- p.122 / Chapter CHAPTER 7： --- General Discussion and Conclusions --- p.135 / BIblography --- p.139

Siu Pak-yan. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2001. / Includes bibliographical references (leaves 158-174). / Abstracts in English and Chinese. / Acknowledgement --- p.i / Abstract --- p.ii / 摘要 --- p.iv / Contents --- p.vi / List of Figures and Tables --- p.xiii / List of Abbreviations --- p.xviii / Chapter Chapter 1 --- Introduction / Chapter 1.1 --- Characteristics of Arsenic Compound --- p.1 / Chapter 1.1.1 --- Arsenic Compounds are Used as Poison --- p.1 / Chapter 1.1.2 --- Arsenic Compounds are Used as Medicine --- p.2 / Chapter 1.2 --- Arsenic Trioxide is a Traditional Chinese Medicine --- p.3 / Chapter 1.3 --- Properties of Arsenic Trioxide --- p.5 / Chapter 1.4 --- Use of Arsenic Trioxide in Cancer Treatment --- p.7 / Chapter 1.4.1 --- Arsenic Trioxide as a Therapeutic Agent in the Treatment of Acute Promyelocytic Leukemia --- p.7 / Chapter 1.4.1.1 --- Characteristics of Acute Promyelocytic Leukemia --- p.7 / Chapter 1.4.1.2 --- Treatment of Acute Promyelocytic Leukemia with All-Trans Retinoic Acid --- p.10 / Chapter 1.4.1.3 --- Treatment of Acute Promyelocytic Leukemia with Arsenic Trioxide --- p.11 / Chapter 1.4.1.4 --- Action Mechanism of Arsenic Trioxide --- p.13 / Chapter 1.4.2 --- Arsenic Trioxide as a Therapeutic Agent in the Treatment of Non-APL Leukemia --- p.15 / Chapter 1.4.3 --- Arsenic Trioxide as a Therapeutic Agent in the Treatment of Solid Tumors --- p.16 / Chapter 1.5 --- Human Hepatocellular Carcinoma --- p.16 / Chapter 1.5.1 --- The Incidence of Liver Cancer --- p.16 / Chapter 1.5.2 --- Classification of Liver Cancer --- p.17 / Chapter 1.6 --- Aim of the Project --- p.17 / Chapter 1.6.1 --- In Vitro Study of the Effect of Arsenic Trioxide on HepG2 Cells --- p.19 / Chapter 1.6.2 --- In Vivo Study of the Effect of Arsenic Trioxide by Tumor-Bearing Nude Mice Model --- p.20 / Chapter 1.6.3 --- "In Vitro Study of the Effect of Arsenic Trioxide on Multidrug-Resistant Human Hepatocellular Carcinoma Cell Line, R-HepG2" --- p.22 / Chapter Chapter 2 --- Materials and Methods / Chapter 2.1 --- Materials --- p.24 / Chapter 2.1.1 --- Cell Lines and Culture Medium --- p.24 / Chapter 2.1.1.1 --- Cell Lines --- p.24 / Chapter 2.1.1.2 --- Culture Medium --- p.25 / Chapter 2.1.2 --- Chemicals --- p.26 / Chapter 2.1.3 --- Reagents and Buffers --- p.27 / Chapter 2.1.3.1 --- Phosphate Buffered Saline (PBS) --- p.27 / Chapter 2.1.3.2 --- "3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide (MTT) Solution" --- p.27 / Chapter 2.1.3.3 --- Reagents for DNA Fragmentation Assay --- p.21 / Chapter 2.1.3.3.1 --- DNA Lysis Buffer --- p.27 / Chapter 2.1.3.3.2 --- Tris-EDTA (TE) Buffer --- p.27 / Chapter 2.1.3.3.3 --- Tris-Acetate (TAE) Buffer --- p.28 / Chapter 2.1.3.3.4 --- Proteinase K and Ribonuclease A (RNase A) --- p.28 / Chapter 2.1.3.3.5 --- 6X DNA Loading Dye --- p.28 / Chapter 2.1.3.3.6 --- One Hundred Base-Pair DNA Ladder --- p.28 / Chapter 2.1.3.4 --- Reagents for Western Blot Analysis --- p.29 / Chapter 2.1.3.4.1 --- SDS Lysis Buffer --- p.29 / Chapter 2.1.3.4.2 --- 4X Lower Gel Buffer --- p.29 / Chapter 2.1.3.4.3 --- 4X Upper Gel Buffer --- p.29 / Chapter 2.1.3.4.4 --- 10X SDS Running Buffer --- p.29 / Chapter 2.1.3.4.5 --- 2X SDS Sample Loading Dye --- p.30 / Chapter 2.1.3.4.6 --- Electroblotting Buffer --- p.30 / Chapter 2.1.3.4.7 --- Tris-Buffered Saline with 01% Tween-20 (TBS-T) --- p.30 / Chapter 2.1.3.4.8 --- Lysis Buffer for Detection of the Release of Cytochrome C --- p.31 / Chapter 2.1.3.5 --- Propidium Iodide (PI) --- p.31 / Chapter 2.1.3.6 --- "5，5 ´ة,6,6´ة-tetrachloro-1,1',3,3 '-tetraethylbenzimidazolyl carbocyanine Iodide (JC-1)" --- p.31 / Chapter 2.1.3.7 --- Reagents for In Vivo Study --- p.32 / Chapter 2.1.3.7.1 --- Saline --- p.32 / Chapter 2.1.3.7.2 --- Homogenizing Buffer --- p.32 / Chapter 2.1.3.7.3 --- 10% Buffered Formalin --- p.32 / Chapter 2.1.3.7.4 --- Acid Alcohol --- p.32 / Chapter 2.1.3.7.5 --- Scott's Tap Water --- p.32 / Chapter 2.1.3.7.6 --- 0.5% Aqueous Eosin --- p.33 / Chapter 2.2 --- Methods --- p.33 / Chapter 2.2.1 --- MTT Assay --- p.33 / Chapter 2.2.2 --- Trypan Blue Exclusion Assay --- p.34 / Chapter 2.2.3 --- Analysis of Cell-Cycle Phase Distribution by Flow Cytometry with PI Staining --- p.34 / Chapter 2.2.4 --- DNA Fragmentation Assay --- p.35 / Chapter 2.2.5 --- Quantification of Apoptosis by Flow Cytometry with Annexin V-PI Staining --- p.36 / Chapter 2.2.6 --- Assessment of the Change in Mitochondrial Membrane Potential (ΔΦm) --- p.37 / Chapter 2.2.7 --- Western Analysis --- p.38 / Chapter 2.2.8 --- Glucose Uptake Assay --- p.40 / Chapter 2.2.9 --- ATP Production Assay --- p.41 / Chapter 2.2.10 --- In Vivo Study --- p.44 / Chapter 2.2.10.1 --- Animal Model --- p.44 / Chapter 2.2.10.2 --- Cell Line --- p.44 / Chapter 2.2.10.3 --- Treatment with Arsenic Trioxide --- p.44 / Chapter 2.2.10.4 --- Assessment of the Anti-Cancer Activity of Arsenic Trioxide --- p.45 / Chapter 2.2.10.5 --- Tissue Sample Preparation --- p.45 / Chapter 2.2.10.5.1 --- Preparation of Plasma --- p.45 / Chapter 2.2.10.5.2 --- Preparation of Liver Tissue Homogenate --- p.46 / Chapter 2.2.10.5.3 --- Preparation of Cytosolic Fraction --- p.46 / Chapter 2.2.10.6 --- Measurement of the Plasma Enzyme Activity --- p.46 / Chapter 2.2.10.6.1 --- "Plasma Creatine Kinase (CK) Activity, Plasma Lactate Dehydrogenase (LDH) Activity, Plasma Alanine Transaminase (ALT) Activity and Plasma Asparate Transaminase (AST) Activity" --- p.46 / Chapter 2.2.10.7 --- Preparation of Tissue for Light Microscopic Study --- p.48 / Chapter 2.2.10.8 --- Measurement of the Basal Reduced Glutathione (GSH) Level of Liver Tissue --- p.51 / Chapter 2.2.10.9 --- "Measurement of the Activity of Antioxidant Enzyme, Glutathione S-Transferase (GST) of Liver Tissue" --- p.53 / Chapter 2.3 --- Statistical Analysis --- p.54 / Chapter Chapter 3 --- "In Vitro Study of Arsenic Trioxide on Acute Promyelocytic Leukemia Cell Line, NB-4" / Chapter 3.1 --- Introduction --- p.55 / Chapter 3.2 --- Principle of Flow Cytometry with Annexin V-PI Staining --- p.56 / Chapter 3.3 --- The Effect of Arsenic Trioxide on Cell Proliferation of NB-4 Cells --- p.59 / Chapter 3.4 --- Study of the Action Mechanism of Arsenic Trioxide upon Treatment of NB-4 Cells --- p.61 / Chapter 3.5 --- Summary --- p.63 / Chapter Chapter 4 --- "In Vitro Study of Arsenic Trioxide on Human Hepatocellular Carcinoma Cell Line, HepG2" / Chapter 4.1 --- Introduction --- p.64 / Chapter 4.2 --- The Effect of Arsenic Trioxide on Cell Proliferation of HepG2 Cells by MTT Assay --- p.66 / Chapter 4.3 --- The Effect of Arsenic Trioxide on HepG2 Cells at Clinically Achievable Concentration --- p.68 / Chapter 4.3.1 --- The Cytotoxicity of Arsenic Trioxide on HepG2 Cells by Trypan Blue Exclusion Assay --- p.68 / Chapter 4.3.2 --- The Effect of Arsenic Trioxide on Cell-Cycle Phase Distribution --- p.71 / Chapter 4.3.3 --- The Underlying Mechanism of the Cytotoxic Effect of Arsenic Trioxide 一 Necrosis or Apoptosis? --- p.74 / Chapter 4.3.3.1 --- DNA Fragmentation Assay --- p.74 / Chapter 4.3.3.2 --- Flow Cytometry with Annexin V-PI Staining --- p.76 / Chapter 4.3.3.3 --- Brief Conclusion --- p.78 / Chapter 4.3.4 --- The Study of the Mechanism of Apoptotic Pathway --- p.78 / Chapter 4.3.4.1 --- Activation of Caspase-3 upon Arsenic Trioxide Treatment --- p.79 / Chapter 4.3.4.2 --- The Participation of Mitochondria in Arsenic Trioxide-Induced Apoptosis --- p.81 / Chapter 4.3.4.2.1 --- The Change in Mitochondrial Membrane Potential upon Arsenic Trioxide Treatment --- p.81 / Chapter 4.3.4.2.2 --- The Study of the Release of Cytochrome C from the Mitochondria to Cytosol upon Treatment with Arsenic Trioxide --- p.85 / Chapter 4.3.4.2.3 --- Brief Conclusion --- p.87 / Chapter 4.4 --- Arsenic Trioxide Mediated Its Effect via Other Action Mechanisms --- p.87 / Chapter 4.4.1 --- The Effect of Arsenic Trioxide on the Expression of Glucose Transporters 1 and2 --- p.88 / Chapter 4.4.2 --- The Effect of Arsenic Trioxide on Glucose Uptake --- p.91 / Chapter 4.4.3 --- The Effect of Arsenic Trioxide on ATP Production --- p.93 / Chapter 4.4.4 --- Brief Conclusion --- p.93 / Chapter 4.5 --- Summary --- p.95 / Chapter Chapter 5 --- In Vivo Study of Arsenic Trioxide on HepG2-Bearing Nude Mice / Chapter 5.1 --- Introduction --- p.96 / Chapter 5.2 --- Treatment with Arsenic Trioxide --- p.97 / Chapter 5.3 --- Assessment of the Anti-Tumor Effect of Arsenic Trioxide --- p.99 / Chapter 5.4 --- The Effect of Arsenic Trioxide toward Normal Tissues --- p.103 / Chapter 5.4.1 --- The Effect of Arsenic Trioxide on Liver --- p.104 / Chapter 5.4.1.1 --- Morphological Study --- p.104 / Chapter 5.4.1.2 --- Enzymatic Study --- p.107 / Chapter 5.4.1.3 --- Brief Conclusion --- p.107 / Chapter 5.4.2 --- The Effect of Arsenic Trioxide on Heart --- p.110 / Chapter 5.4.2.1 --- Morphological Study --- p.110 / Chapter 5.4.2.2 --- Enzymatic Study --- p.112 / Chapter 5.4.2.3 --- Brief Conclusion --- p.112 / Chapter 5.5 --- Involvement of the Glutathione Redox System --- p.115 / Chapter 5.5.1 --- Basal GSH Level --- p.115 / Chapter 5.5.2 --- The Activity of Glutathion S-Transferase --- p.117 / Chapter 5.5.3 --- Brief Conclusion --- p.117 / Chapter 5.6 --- Summary --- p.120 / Chapter Chapter 6 --- "In Vitro Study of Arsenic Trioxide on Multidrug-Resistant Human Hepatocellular Carcinoma Cell Line, R-HepG2" / Chapter 6.1 --- Introduction --- p.121 / Chapter 6.2 --- The Effect of Doxorubicin on the Parental HepG2 Cells and R-HepG2 Cells by MTT Assay --- p.123 / Chapter 6.3 --- The Effect of Arsenic Trioxide on Cell Proliferation of R-HepG2 Cells by MTT Assay --- p.126 / Chapter 6.4 --- The Effect of Arsenic Trioxide on Cell-Cycle Phase Distribution of R-HepG2 Cells --- p.129 / Chapter 6.5 --- Trioxide on R-HepG2 Cells ´ؤ Necrosis or Apoptosis? --- p.131 / Chapter 6.5.1 --- DNA Fragmentation Assay --- p.131 / Chapter 6.5.2 --- Flow Cytometry with Annexin V-PI Staining --- p.133 / Chapter 6.5.3 --- Brief Conclusion --- p.133 / Chapter 6.6 --- Examination of the Probable Involvement of Arsenic Trioxide as a Substrate of P-Glycoprotein --- p.135 / Chapter 6.7 --- Summary --- p.137 / Chapter Chapter 7 --- Discussion / Chapter 7.1 --- The Significance of the Study of Arsenic Trioxide in the Treatment of Arsenic Trioxide --- p.138 / Chapter 7.2 --- Comparison of Preparation of Drug in Present Study with Others --- p.140 / Chapter 7.3 --- Effect of Arsenic Trioxide on Human Hepatocellular Carcinoma --- p.142 / Chapter 7.4 --- Mechanism Study of Arsenic Trioxide --- p.142 / Chapter 7.5 --- Dosage of Arsenic Trioxide Used in In Vivo Study --- p.152 / Chapter 7.6 --- Cytotoxicity of Arsenic Trioxide toward Normal Tissues --- p.153 / Chapter 7.7 --- "Effect of Arsenic Trioxide on Multidrug-Resistant Human Hepatocellular Carcinoma Cell Line, R-HepG2" --- p.154 / Chapter 7.8 --- Conclusions and Future Prospect --- p.156 / Chapter Chapter 8 --- References / Chapter 8.1 --- English References --- p.158 / Chapter 8.2 --- Chinese References --- p.174 / Chapter 8.3 --- Online References --- p.174

by Chow Ka Yee. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2002. / Includes bibliographical references (leaves 203-221). / Abstracts in English and Chinese. / Acknowledgements --- p.i / Abstract --- p.ii / Abstract in Chinese --- p.iv / List of Abbreviations --- p.vi / Table of Contents --- p.xi / List of Figures --- p.xviii / List of Tables --- p.xxii / Chapter CHAPTER 1 --- INTRODUCTION --- p.1 / Chapter 1.1 --- The Characteristics of Arsenic Trioxide (AS2O3) --- p.2 / Chapter 1.2 --- The Therapeutic Applications of Arsenic Trioxide (As203) --- p.5 / Chapter 1.3 --- Acute Promyelocytic Leukemia (APL) --- p.6 / Chapter 1.3.1 --- Pathologies of APL --- p.7 / Chapter 1.3.2 --- All Trans Retinoic Acid (ATRA) Treatment of APL Patients --- p.7 / Chapter 1.3.3 --- Clinical Trials of Arsenic Trioxide (As203) on APL Patients --- p.9 / Chapter 1.3.4 --- In Vitro and In Vivo Studies of Arsenic Trioxide (As203) in the Treatment of APL --- p.10 / Chapter 1.3.4.1 --- Induction of Apoptosis --- p.11 / Chapter 1.3.4.2 --- Induction of Cell Differentiation --- p.11 / Chapter 1.3.5 --- General Toxicity and Side Effects of Arsenic Trioxide (AS2O3) on APL Patients --- p.12 / Chapter 1.4 --- Effects of Arsenic Trioxide (As203) on Other Primary Cancer Cells and Cancer Cell Lines --- p.12 / Chapter 1.5 --- Epidemiology of Breast Cancer --- p.14 / Chapter 1.6 --- Classification of Breast Cancer --- p.17 / Chapter 1.7 --- Etiology of Breast Cancer --- p.17 / Chapter 1.8 --- Hormones and Breast Cancer --- p.18 / Chapter 1.9 --- Estrogen Receptors (ER) --- p.20 / Chapter 1.9.1 --- Structures of Estrogen Receptors (ER) --- p.21 / Chapter 1.9.2 --- Estrogen Receptors (ER) Mediated Signaling Pathway --- p.22 / Chapter 1.9.2.1 --- Ligand Dependent Pathway --- p.22 / Chapter 1.9.2.2 --- Ligand Independent Pathway --- p.22 / Chapter 1.9.2.3 --- Estrogen Response Element (ERE)-Independent Pathway --- p.23 / Chapter 1.9.2.4 --- Non-Genomic Pathway --- p.23 / Chapter 1.9.3 --- Estrogen Receptors (ER) Regulated Gene Expression --- p.25 / Chapter 1.10 --- Current Therapy of Breast Cancer --- p.26 / Chapter 1.10.1 --- Hormonal Therapy (Anti-Estrogenicity) --- p.26 / Chapter 1.10.1.1 --- Tamoxifen --- p.26 / Chapter 1.10.1.2 --- Other Pure Anti-Estrogens --- p.28 / Chapter 1.10.2 --- Regulation of Estrogen Receptors (ER) and Transcription Coregulators --- p.29 / Chapter 1.10.3 --- Apoptosis Induction --- p.29 / Chapter 1.11 --- Aims of Study --- p.30 / Chapter CHAPTER 2 --- MATERIALS AND METHODS --- p.32 / Chapter 2.1 --- Materials --- p.33 / Chapter 2.1.1 --- Cell Lines and Culture Media --- p.33 / Chapter 2.1.1.1 --- Cell Lines --- p.33 / Chapter 2.1.1.2 --- Culture Media --- p.34 / Chapter 2.1.2 --- Chemicals --- p.35 / Chapter 2.1.3 --- Reagents and Buffers --- p.36 / Chapter 2.1.3.1 --- Reagents for MTT Assay --- p.36 / Chapter 2.1.3.2 --- Reagents for [methyl-3H] Thymidine Incorporation into DNA --- p.37 / Chapter 2.1.3.3 --- Reagents for Trypan Blue Exclusion Assay --- p.37 / Chapter 2.1.3.4 --- Reagents and Buffers for Western Blot Analysis --- p.37 / Chapter 2.1.3.5 --- Reagents and Buffers for Flow Cytometry --- p.40 / Chapter 2.1.3.6 --- Reagents and Buffers Reverse Transcription Polymerase Chain Reaction (RT-PCR) --- p.40 / Chapter 2.1.3.7 --- Reagents for Transfection and Luciferase Reporter Assay --- p.41 / Chapter 2.1.3.8 --- Reagents and Buffers for In Vivo Studies --- p.42 / Chapter 2.2 --- Methods --- p.42 / Chapter 2.2.1 --- In Vitro Studies --- p.42 / Chapter 2.2.1.1 --- Cell Treatment --- p.42 / Chapter 2.2.1.2 --- Drug Preparation --- p.43 / Chapter 2.2.1.3 --- MTT Assay --- p.43 / Chapter 2.2.14 --- Trypan Blue Exclusion Assay --- p.44 / Chapter 2.2.1.5 --- [methyl-3H] Thymidine Incorporation into DNA --- p.45 / Chapter 2.2.1.6 --- Detection of DNA Fragmentation --- p.45 / Chapter 2.2.1.7 --- ERα Competitive Binding Assay --- p.47 / Chapter 2.2.1.8 --- Cell Cycle Analysis by Flow Cytometry with Propidium Iodide (PI) Staining --- p.48 / Chapter 2.2.1.9 --- Cell Cycle Analysis by Flow Cytometry with Annexin V-PI Staining --- p.48 / Chapter 2.2.1.10 --- Cell Cycle Analysis by Flow Cytometry with JC-1 Staining --- p.49 / Chapter 2.2.1.11 --- Cell Cycle Analysis by Flow Cytometry with Hydroethidine (HE) Staining --- p.50 / Chapter 2.2.1.12 --- Western Blot Analysis of Proteins --- p.50 / Chapter 2.2.1.13 --- Assessment of the Transcriptional Activity of ERα --- p.55 / Chapter 3.2.1.14 --- Reverse Transcription Polymerase Chain Reaction (RT-PCR) --- p.57 / Chapter 2.2.2 --- In Vivo Studies --- p.61 / Chapter 2.2.2.1 --- Animal Models --- p.61 / Chapter 2.2.2.2 --- Treatment Schedules --- p.61 / Chapter 2.2.2.3 --- Sacrifice of Nude Mice --- p.61 / Chapter 2.2.2.4 --- Enzymatic Assays --- p.62 / Chapter 2.2.2.4.1 --- Aspartate Transaminase (AST) --- p.63 / Chapter 2.2.2.4.2 --- Alanine Transaminase (ALT) --- p.64 / Chapter 2.2.2.4.3 --- Creatine Kinase (CK) --- p.65 / Chapter 2.2.2.4.4 --- Lactate Dehydrogenase (LDH) --- p.66 / Chapter CHAPTER 3 --- "Effects of Arsenic Trioxide (As203) on Human Breast Adenocarcinoma Cell Line, MCF-7 Cell Line" --- p.68 / Chapter 3.1 --- Introduction --- p.69 / Chapter 3.2 --- Effect of As203 on Cell Survival of MCF-7 cells by MTT Assay --- p.70 / Chapter 3.3 --- Cytotoxicity of As203 on MCF-7 Cells by Trypan Blue Exclusion Assay --- p.72 / Chapter 3.4 --- Effect of As203 on DNA Synthesis and Cell Proliferation of MCF-7 cells by [methyl-3H] Thymidine Incorporation into DNA --- p.76 / Chapter 3.5 --- Comparison of Cytotoxicity of AS2O3 on MCF-7 Cells with that of Tamoxifen --- p.79 / Chapter 3.6 --- Summary --- p.82 / Chapter CHAPTER 4 --- Effects of Arsenic Trioxide (As203) on 17β Estradiol Stimulated MCF-7 cells --- p.83 / Chapter 4.1 --- Introduction --- p.84 / Chapter 4.2 --- Effect of 17β estradiol on Cell Viability of MCF-7 Cells by MTT Assay --- p.86 / Chapter 4.3 --- Effect of As203 and 17β Estradiol on Cell Survival of MCF-7 Cells by MTT Assay --- p.88 / Chapter 4.4 --- Cytotoxicity of As203 on 17β Estradiol Stimulated MCF-7 cells by Cell Number Counting with Hemacytometer --- p.92 / Chapter 4.5 --- Growth Inhibitory Effect of As203 on 17β Estradiol stimulated MCF-7 cells by [methyl-3H] Thymidine Incorporation into DNA --- p.94 / Chapter 4.6 --- "Effect of As203 on Cell Survival of Hormone Independent Breast Cancer Cell Line, MDA-MB-231 Cells" --- p.96 / Chapter 4.7 --- Summary --- p.100 / Chapter CHAPTER 5 --- Effects of Arsenic Trioxide (As203) on Normal Cells --- p.102 / Chapter 5.1 --- Introduction --- p.103 / Chapter 5.2 --- "Effect of As203 on Normal Human Fibroblast Cell Line, Hs68" --- p.104 / Chapter 5.3 --- Effects of As203 on the Normal Cells of Nude Mice --- p.106 / Chapter 5.3.1 --- Effect of AS2O3 on Aspartate Transaminase (AST) Activity of Nude Mice --- p.107 / Chapter 5.3.2 --- Effect of As203 on Alanine Transaminase (ALT) Activity of Nude Mice --- p.109 / Chapter 5.3.3 --- Effect of As203 on Creatine Kinase (CK) Activity of Nude Mice TABLE OF CONTENTS --- p.111 / Chapter 5.3.4 --- Effect of As203 on Lactate Dehydrogenase (LDH) Activity of Nude Mice --- p.113 / Chapter 5.4 --- Summary --- p.115 / Chapter CHAPTER 6 --- Action Mechanisms underlying the Survival Inhibitory Effects of Arsenic Trioxide (As203) on MCF-7 cells --- p.116 / Chapter 6.1 --- Introduction --- p.117 / Chapter 6.2 --- Detection of Apoptosis --- p.119 / Chapter 6.2.1 --- Detection of DNA Fragmentation --- p.119 / Chapter 6.2.2 --- Phosphatidylserine (PS) Externalization Detected by Flow Cytometry with Annexin V-PI Staining --- p.124 / Chapter 6.2.2.1 --- The Principle --- p.124 / Chapter 6.2.2.2 --- PS Externalization upon AS2O3 Treatment --- p.126 / Chapter 6.3 --- Analysis of Cell Cycle Distribution of MCF-7 Cells --- p.130 / Chapter 6.3.1 --- The Principle --- p.130 / Chapter 6.3.2 --- Regulation of Cell Cycle Distribution of MCF-7 Cells upon As2O3 Treatment --- p.131 / Chapter 6.4 --- The Action Mechanisms Underlying As203 Induced Apoptosis or Cell Cycle Arrest --- p.137 / Chapter 6.4.1 --- Effect of As203 on Mitochondrial Membrane Potential of MCF-7 Cells --- p.137 / Chapter 6.4.2 --- Regulation of Free Oxidative Species (ROS) Production in MCF-7 Cells upon AS2O3 Treatment --- p.140 / Chapter 6.4.2.1 --- Analysis of Superoxide Production in MCF-7 Cells upon AS2O3 Treatment by Flow Cytometry with Hydroethidine (HE) Staining --- p.140 / Chapter 6.4.2.2 --- Effect of As203 on Cell Survival of MCF-7 Cells Co-treated with N-Acteyl-L-Cysteine (NAC) by MTT Assay --- p.143 / Chapter 6.4.3 --- Regulation of Bcl-2 Protein Level in MCF-7 Cells upon As2O3 Treatment --- p.145 / Chapter 6.4.4 --- Regulation of p53 Protein Level in MCF-7 Cells upon AS2O3 Treatment --- p.147 / Chapter 6.5 --- Summary --- p.149 / Chapter CHAPTER 7 --- Effects of Arsenic Trioxide (As203) on Estrogen Receptor a (ERα) Mediated Signaling Pathway in MCF-7 cells --- p.150 / Chapter 7.1 --- Introduction --- p.151 / Chapter 7.2 --- Effect of As203 on Estrogen Binding to Estrogen Receptor a (ERα) by ERα Competitive Binding Assay --- p.152 / Chapter 7.3 --- Regulation of Estrogen Receptor a (ERα) mRNA Level upon As2O3 Treatment by RT-PCR --- p.156 / Chapter 7.4 --- Regulation of Estrogen Receptor a (ERα) Protein Level upon As2O3 Treatment --- p.159 / Chapter 7.5 --- Regulation of Estrogen Receptor a (ERα) Transcriptional Activity upon AS2O3 treatment --- p.161 / Chapter 7.6 --- "Regulation of Estrogen Target Gene, c-myc, Protein Level upon As2O3 Treatment" --- p.164 / Chapter 7.7 --- Effects of As203 on Cell Cycle Distribution of MCF-7 Cells under Estrogens Stimulation --- p.167 / Chapter 7.8 --- Summary --- p.173 / Chapter CHAPTER 8 --- Discussion --- p.174 / Chapter 8.1 --- The Anti-Tumor Effects of As203 on MCF-7 Cells --- p.175 / Chapter 8.2 --- Cytotoxicity of As203 on MCF-7 Cells --- p.175 / Chapter 8.2.1 --- Induction of Apoptosis in MCF-7 Cells upon As2〇3 Treatment --- p.176 / Chapter 8.2.2 --- Action Mechanisms Underlying the Induction of Apoptosis by As2〇3 --- p.178 / Chapter 8.3 --- Growth Inhibition of As203 on MCF-7 Cells --- p.182 / Chapter 8.3.1 --- Cell Cycle Regulation of MCF-7 Cells upon As203 Treatment --- p.182 / Chapter 8.4 --- Growth Inhibitory Effects of As203 on Estrogen Stimulated MCF-7 Cells --- p.186 / Chapter 8.4.1 --- Regulation of Estrogen Receptor a (ERα) Signaling Pathway in MCF-7 cells upon as2o3 Treatment --- p.188 / Chapter 8.5 --- Cross Talk of ERα Signaling Pathway and Apoptosis in Mediating the Anti-Tumor Effects of As203 on MCF-7 Cells --- p.195 / Chapter 8.6 --- Toxicity of AS2O3 towards Normal Tissues --- p.197 / Chapter CHAPTER 9 --- Conclusion and Future Perspectives --- p.200 / Chapter 9.1 --- Conclusion --- p.200 / Chapter 9.2 --- Future Perspectives --- p.202 / References --- p.203

Zhou, Linli. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2007. / Includes bibliographical references (leaves 108-118). / Abstracts in English and Chinese. / Acknowledgements --- p.i / Abstract --- p.ii / 論文摘要 --- p.iv / Abbreviations --- p.v / List of Figures --- p.vii / List of Tables --- p.ix / Table of Contents --- p.x / Chapter Chapter 1 --- Introduction --- p.1 / Chapter 1.1 --- Breast Cancer --- p.1 / Chapter 1.1.1 --- Introduction to Breast Cancer --- p.1 / Chapter 1.1.2 --- Types of Breast Cancer --- p.3 / Chapter 1.1.3 --- Epidemiologic Risk Factors and Etiology --- p.4 / Chapter 1.2 --- Estrogen and Breast Cancer --- p.7 / Chapter 1.3 --- Estrogen Receptor --- p.9 / Chapter 1.4 --- Current Treatment of Breast Cancer --- p.10 / Chapter 1.4.1 --- Chemotherapy --- p.10 / Chapter 1.4.2 --- Hormonal (Anti-Estrogen) Therapy --- p.11 / Chapter 1.4.2.1 --- Tamoxifen and Other Anti-estrogens --- p.12 / Chapter 1.4.2.2 --- Disadvantages of Tamoxifen --- p.13 / Chapter 1.5 --- Arsenic Trioxide --- p.14 / Chapter 1.5.1 --- The Characteristics of Arsenic Trioxide (AS2O3) --- p.14 / Chapter 1.5.2 --- The Medical use of Arsenic Trioxide (As2O3) --- p.16 / Chapter 1.5.3 --- Arsenic Trioxide (As2O3) in treating Acute Promyelocytic Leukemia (APL) --- p.17 / Chapter 1.5.3.1 --- Acute Promyelocytic Leukemia (APL) --- p.17 / Chapter 1.5.3.2 --- All-trans Retinoic Acid (ATRA) Treatment of APL --- p.18 / Chapter 1.5.3.3 --- Clinical Trial of the Arsenic Trioxide on APL --- p.19 / Chapter 1.5.3.4 --- In vitro and in vivo Study of Arsenic Trioxide (As2O3) in treating APL --- p.19 / Chapter 1.5.3.5 --- Common Side Effects of Arsenic Trioxide (As2O3) on APL --- p.21 / Chapter 1.5.4 --- Anti-cancer effect of Arsenic Trioxide on other cancers --- p.23 / Chapter 1.6 --- Aim of Study --- p.24 / Chapter Chapter 2 --- Materials and Methods --- p.26 / Chapter 2.1 --- Materials --- p.27 / Chapter 2.1.1 --- Cell Lines and Culture Medium --- p.27 / Chapter 2.1.1.1 --- Cell Lines --- p.27 / Chapter 2.1.1.2 --- Culture Medium --- p.27 / Chapter 2.1.2 --- Chemicals --- p.28 / Chapter 2.1.3 --- Buffers and Reagents --- p.29 / Chapter 2.1.4 --- Reagents for MTT Assay --- p.30 / Chapter 2.1.5 --- Reagents for DNA Fragmentation --- p.31 / Chapter 2.1.5.1 --- Reagents for DNA Extraction --- p.31 / Chapter 2.1.5.2 --- Reagents for Gel Electrophoresis --- p.31 / Chapter 2.1.6 --- Reagents for Western Blotting --- p.32 / Chapter 2.1.6.1 --- Reagents for Protein Extraction --- p.32 / Chapter 2.1.6.2 --- Reagents for SDS-PAGE --- p.33 / Chapter 2.1.7 --- Reagents for Flow Cytometry --- p.36 / Chapter 2.1.8 --- In Vivo Study --- p.37 / Chapter 2.2 --- Methods --- p.38 / Chapter 2.2.1 --- Cell Treatment --- p.38 / Chapter 2.2.2 --- Trypan Blue Exclusion Assay --- p.38 / Chapter 2.2.3 --- MTT Assay --- p.38 / Chapter 2.2.4 --- Detection of DNA Fragmentation --- p.39 / Chapter 2.2.5 --- Flow Cytometry --- p.40 / Chapter 2.2.5.1 --- Detection of Cell Cycle Pattern with PI --- p.40 / Chapter 2.2.5.2 --- Detection of Apoptosis with Annexin V-PI --- p.40 / Chapter 2.2.6 --- Western Blot Analysis --- p.41 / Chapter 2.2.6.1 --- Protein Extraction --- p.41 / Chapter 2.2.6.2 --- Protein Concentration Determination --- p.41 / Chapter 2.2.6.3 --- Western Blotting --- p.42 / Chapter 2.2.7 --- In Vivo Study --- p.44 / Chapter 2.2.7.1 --- Animal Model --- p.44 / Chapter 2.2.7.2 --- Treatment Schedule --- p.44 / Chapter 2.2.7.3 --- Toxicity of Arsenic Trioxide --- p.45 / Chapter Chapter 3 --- Anti-Proliferation Effect of As2O3 on MDA-MB-231 cells --- p.47 / Chapter 3.1 --- Study the Anti-proliferation Effect of As2O3 on MDA-MB-231 Cells by MTT Assay --- p.48 / Chapter 3.2 --- Comparsion Anti-proliferation Effect of AS2O3 on MDA-MB-231 Cells to that of Tamoxifen --- p.50 / Chapter 3.3 --- "Study Toxicity of AS2O3 on Normal Breast Cells Line, 184B5" --- p.52 / Chapter 3.4 --- Summary --- p.54 / Chapter Chapter 4 --- Mechanism of Growth Inhibition Effect of As2O3 on MDA-MB-231 cells --- p.56 / Chapter 4.1 --- Cell Cycle Analysis of As2O3 Treated MDA-MB-231 Cells --- p.57 / Chapter 4.2 --- Detection of DNA Fragmentation --- p.60 / Chapter 4.3 --- Detection of Apoptosis Induced by AS2O3 on MDA-MB-231 Cells by Flow Cytometry --- p.62 / Chapter 4.4 --- Regulation of Apoptotic Related Protein by As2O3 on MDA-MB-231 Cells --- p.64 / Chapter 4.4.1 --- Expression Level of Bcl-2 and Bax Protein --- p.66 / Chapter 4.4.2 --- Expression Level of Cytochrome C --- p.69 / Chapter 4.4.3 --- Expression Level of Caspase9 --- p.71 / Chapter 4.4.4 --- Expression Level of FasL --- p.73 / Chapter 4.4.5 --- Expression Level of Caspase8 --- p.75 / Chapter 4.4.6 --- Expression Level of Caspase3 --- p.77 / Chapter 4.4.7 --- Expression Level of Poly (ADP-ribose) Polymerase (PARP) --- p.79 / Chapter 4.4.8 --- Expression Level of p53 --- p.81 / Chapter 4.5 --- Regulation of Cell Cycle Related Protein by AS2O3 on MDA-MB-231 Cells --- p.83 / Chapter 4.5.1 --- Expression Level of Cyclin B --- p.84 / Chapter 4.5.2 --- Expression Level of Cyclin E --- p.86 / Chapter 4.6 --- Summary --- p.88 / Chapter Chapter 5 --- In Vivo Study of Anti-tumor Effect of As2O3 --- p.89 / Chapter 5.1 --- Anti-tumor Effect of AS2O3 on Tumor Bearing Nude Mice --- p.90 / Chapter 5.2 --- Toxic Effect of AS2O3 on Normal Tissues --- p.93 / Chapter 5.3 --- Summary --- p.98 / Chapter Chapter 6 --- Discussion --- p.99 / Chapter 6.1 --- Anti-tumor Effect of AS2O3 on Breast Cancer --- p.100 / Chapter 6.2 --- Induction of Apoptosis and Cell Cycle arrest by AS2O3 --- p.101 / Chapter 6.3 --- Side Effect of AS2O3 on Breast Cancer Treatment --- p.103 / Chapter Chapter 7 --- Future Perspectives --- p.105 / Chapter 7.1 --- Future Perspectives --- p.106 / References --- p.108