Chan, Hoi Ling. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2008. / Includes bibliographical references (leaves 206-220). / Abstracts in English and Chinese. / ACKNOWLEDGEMENTS --- p.i / ABBREVIATIONS --- p.ii / ABSTRACT --- p.viii / 撮要 --- p.xii / PUBLICATIONS --- p.xiv / TABLE OF CONTENTS --- p.xv / Chapter CHAPTER 1: --- GENERAL INTRODUCTION / Chapter 1.1 --- Hematopoiesis & Leukemia --- p.1 / Chapter 1.1.1 --- An Overview on Hematopoiesis Development --- p.1 / Chapter 1.1.2 --- Leukemia --- p.6 / Chapter 1.1.2.1 --- General Symptoms of Leukemia --- p.7 / Chapter 1.1.2.2 --- Classification of Leukemia --- p.8 / Chapter 1.1.2.3 --- Conventional Treatment against Leukemia --- p.15 / Chapter 1.1.2.4 --- Novel Approaches --- p.19 / Chapter 1.2 --- The Chinese Medicinal Herb-Banlangen (板藍根） --- p.24 / Chapter 1.2.1 --- An Overview on Natural Indigo Compounds Derived from Banlangen --- p.24 / Chapter 1.2.2 --- Tryptanthrin --- p.29 / Chapter 1.2.2.1 --- Anti-bacterial Activity of Tryptanthrin --- p.29 / Chapter 1.2.2.2 --- Anti-tumor Activity of Tryptanthrin --- p.31 / Chapter 1.2.2.3 --- Anti-inflammatory Activity of Tryptanthrin --- p.33 / Chapter 1.2.2.4 --- Cutting Edges of Tryptanthrin as a Drug --- p.34 / Chapter 1.2.2.5 --- Metabolism of Tryptanthrin --- p.35 / Chapter 1.3 --- Aims and Scopes of This Investigation --- p.37 / Chapter CHAPTER 2: --- MATERIALS AND METHODS / Chapter 2.1 --- Materials --- p.39 / Chapter 2.1.1 --- Animals --- p.39 / Chapter 2.1.2 --- Cell Lines --- p.39 / Chapter 2.1.3 --- "Cell Culture Medium, Buffers and Other Reagents" --- p.41 / Chapter 2.1.4 --- Reagents for 3H-Thymidine Incorporation Assay --- p.45 / Chapter 2.1.5 --- Reagents and Buffers for Flow Cytometry --- p.46 / Chapter 2.1.6 --- Reagents for DNA Extraction --- p.49 / Chapter 2.1.7 --- Reagents for Measuring Caspase Activity --- p.50 / Chapter 2.1.8 --- Reagents for Total RNA Isolation --- p.53 / Chapter 2.1.9 --- Reagents and Buffers for Reversed Transcription-PCR --- p.54 / Chapter 2.1.10 --- Reagents and Buffers for Real Time-PCR --- p.59 / Chapter 2.1.11 --- Reagents and Buffers for Gel Electrophoresis of Nucleic Acids --- p.59 / Chapter 2.1.12 --- "Reagents, Buffers and Materials for Western Blot Analysis" --- p.61 / Chapter 2.2 --- Methods --- p.70 / Chapter 2.2.1 --- Culture of the Tumor Cell Lines --- p.70 / Chapter 2.2.2 --- "Isolation, Preparation and Culture of Mouse Peritoneal Macrophages" --- p.70 / Chapter 2.2.3 --- Determination of Cell Viability --- p.71 / Chapter 2.2.4 --- Determination of Cell Proliferation by [3H]-TdR Incorporation Assay --- p.72 / Chapter 2.2.5 --- Determination of Anti-leukemia Activity In Vivo --- p.73 / Chapter 2.2.6 --- Analysis of Cell Cycle Profile/DNA Content by Flow Cytometry --- p.74 / Chapter 2.2.7 --- Measurement of Apoptosis --- p.75 / Chapter 2.2.8 --- Determination of the Mitochondrial Membrane Potential --- p.77 / Chapter 2.2.9 --- Measurement of Caspase Activity --- p.78 / Chapter 2.2.10 --- Study of Intracellular Accumulation of Reactive Oxygen Species --- p.79 / Chapter 2.2.11 --- Gene Expression Study --- p.80 / Chapter 2.2.12 --- Protein Expression Study --- p.83 / Chapter 2.2.13 --- Measurement of Cell Differentiation --- p.87 / Chapter CHAPTER 3: --- STUDIES ON THE ANTI-PROLIFERATIVE EFFECT OF TRYPTANTHRIN AND INDIRUBIN-3'-OXIME ON MYELOID LEUKEMIA CELLS / Chapter 3.1 --- Introduction --- p.90 / Chapter 3.2 --- Results --- p.94 / Chapter 3.2.1 --- Effects of Indirubin-3'-oxime and Tryptanthrin on the Proliferation of Myeloid Leukemia Cell Lines of Human and Murine Origins In Vitro --- p.94 / Chapter 3.2.2 --- Kinetic and Reversibility Studies of the Anti-proliferative Effect of Tryptanthrin on Murine Myelomonocytic Leukemia WEHI-3B JCS Cells In Vitro --- p.108 / Chapter 3.2.3 --- Cytotoxic Effect of Tryptanthrin on Murine Myelomonocytic Leukemia WEHI-3B JCS Cells In Vitro --- p.113 / Chapter 3.2.4 --- Cytotoxicity of Tryptanthrin on Non-Cancer Cell Line and Primary Myeloid Cells In Vitro --- p.115 / Chapter 3.2.5 --- Effects of Tryptanthrin on the Cell Cycle Profile of the Murine Myelomonocytic Leukemia WEHI-3B JCS Cells In Vitro --- p.118 / Chapter 3.2.6 --- Effects of Tryptanthrin on the Expression of Cell Cycle Related Genes in Murine Myelomonocytic Leukemia WEHI-3B JCS Cells In Vitro --- p.123 / Chapter 3.2.7 --- Expression of CDK-inhibitors in Tryptanthrin-treated Murine Myeloid Leukemia WEHI-3B JCS Cells --- p.126 / Chapter 3.2.8 --- Effects of Tryptanthrin on the In Vivo Tumorigenicity of the Murine Myelomonocytic Leukemia WEHI-3B JCS Cells --- p.128 / Chapter 3.2.9 --- In Vivo Anti-tumor Effect of Tryptanthrin on Murine Myelomonocytic Leukemia WEHI-3B JCS Cells --- p.130 / Chapter 3.3 --- Discussion --- p.132 / Chapter CHAPTER 4: --- STUDIES ON THE APOPTOSIS-INDUCING EFFECT OF TRYPTANTHRIN ON MURINE MYELOMONOCYTIC LEUKEMIA WEHI-3B JCS CELLS / Chapter 4.1 --- Introduction --- p.139 / Chapter 4.2 --- Results --- p.143 / Chapter 4.2.1 --- Induction of DNA Fragmentation by Tryptanthrin in the Murine Myelomonocytic Leukemia WEHI-3B Cells In Vitro --- p.143 / Chapter 4.2.2 --- Induction of Phosphatidylserine Externalization by Tryptanthrin in Murine Myelomonocytic Leukemia WEHI-3B JCS Cells --- p.145 / Chapter 4.2.3 --- Change of Mitochondrial Membrane Potential of Tryptanthrin- treated Murine Myelomonocytic Leukemia WEHI-3B JCS Cells --- p.147 / Chapter 4.2.4 --- Induction of Caspase Activity in Tryptanthrin-treated Murine Myelomonocytic Leukemia WEHI-3B JCS cells --- p.150 / Chapter 4.2.5 --- Induction of Reactive Oxygen Species in Tryptanthrin-treated Murine Myelomonocytic Leukemia WEHI-3B JCS cells --- p.155 / Chapter 4.2.6 --- Expression of Bcl-2 Family Proteins in the Tryptanthrin-treated Murine Myelomonocytic Leukemia WEHI-3B JCS cells --- p.160 / Chapter 4.2.7 --- Effects of Tryptanthrin on the mRNA Expression of Bcl-2 Family Proteins in Murine Myelomonocytic Leukemia WEHI-3B JCS Cells --- p.163 / Chapter 4.2.8 --- Expression of Fas and Fas Ligand Proteins in Tryptanthrin-treated Murine Myelomonocytic Leukemia WEHI-3B JCS cells --- p.167 / Chapter 4.2.9 --- Expression of Pro-Apoptotic Protein in Tryptanthrin- treated Murine Myelomonocytic Leukemia WEHI-3B JCS cells --- p.170 / Chapter 4.2 --- Discussion --- p.173 / Chapter CHAPTER 5: --- STUDIES ON THE DIFFERENTIATION-INDUCING ABILITY OF TRYPTANTHRIN ON MURINE MYELOMONOCYTIC LEUKEMIA WEHI-3B JCS CELLS / Chapter 5.1 --- Introduction --- p.184 / Chapter 5.2 --- Results --- p.186 / Chapter 5.2.1 --- Morphological Studies on Tryptanthrin-treated Murine Myelomonocytic Leukemia WEHI-3B JCS Cells --- p.186 / Chapter 5.2.2 --- Effects of Tryptanthrin on the Cell Size and Granularity of the Murine Myelomonocytic Leukemia WEHI-3B JCS Cells --- p.189 / Chapter 5.2.3 --- Effects of Tryptanthrin on Induction of NBT-reducing Activity in the Murine Myelomonocytic Leukemia WEHI-3B JCS Cells --- p.191 / Chapter 5.3 --- Discussion --- p.195 / Chapter CHAPTER 6: --- CONCLUSIONS AND FUTURE PERSPECTIVES --- p.198 / REFERENCES --- p.206