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Effects of 10-hydroxy-camptothecin and etoposide phosphate on the proliferation, differentiation and survival of the murine myeloid leukemia WEHI-3B JCS cells.

Chan Wai Sing. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2003. / Includes bibliographical references (leaves 199-210). / Abstracts in English and Chinese. / ACKNOWLEDGEMENTS --- p.i / ABBREVIATIONS --- p.ii / ABSTRACT --- p.vi / CHINESE ABSTRACT --- p.x / TABLE OF CONTENTS --- p.xiii / Chapter CHAPTER 1: --- GENERAL INTRODUCTION / Chapter 1.1 --- What is hematopoiesis? --- p.1 / Chapter 1.1.1 --- The development of hematopoietic progenitor cells --- p.1 / Chapter 1.1.2 --- The regulation of hematopoiesis by environmental factors --- p.4 / Chapter 1.1.3 --- The regulation of hematopoiesis by transcription factors --- p.7 / Chapter 1.2 --- Leukemia --- p.9 / Chapter 1.2.1 --- Classification of leukemia --- p.11 / Chapter 1.2.2 --- Pathology and etiology of leukemia --- p.15 / Chapter 1.2.2.1 --- Inheritance --- p.15 / Chapter 1.2.2.2 --- Environmental factors --- p.16 / Chapter 1.2.2.3 --- Virus infection --- p.17 / Chapter 1.2.3 --- Genetics of leukemia --- p.17 / Chapter 1.2.3.1 --- Point mutations --- p.17 / Chapter 1.2.3.2 --- Translocations --- p.18 / Chapter 1.2.3.3 --- Gene and chromosomal deletions --- p.18 / Chapter 1.2.3.4 --- Chromosomal duplication or gene amplification --- p.18 / Chapter 1.2.4 --- Current therapeutic strategies for leukemia --- p.20 / Chapter 1.2.4.1 --- Chemotherapy --- p.20 / Chapter 1.2.4.2 --- Stem cell transplantation --- p.21 / Chapter 1.2.4.3 --- Immunotherapy --- p.22 / Chapter 1.2.4.4 --- Gene therapy --- p.23 / Chapter 1.2.5 --- Novel approaches for the treatment of leukemia --- p.23 / Chapter 1.2.5.1 --- Differentiation therapy of leukemia --- p.24 / Chapter 1.2.5.2 --- Induction of apoptosis in the treatment of leukemia --- p.25 / Chapter 1.3 --- Topoisomerase-targeting agents --- p.27 / Chapter 1.3.1 --- What is topoisomerase? --- p.27 / Chapter 1.3.2 --- Structures and action mechanisms of Top I- and Top II-targeting agents --- p.28 / Chapter 1.3.3 --- Anti-tumor activities of topoisomerase-targeting agents --- p.37 / Chapter 1.4 --- Aims and scopes of this investigation --- p.39 / Chapter CHAPTER 2: --- MATERIALS AND METHODS / Chapter 2.1 --- Materials --- p.42 / Chapter 2.1.1 --- Mice --- p.42 / Chapter 2.1.2 --- Cell lines --- p.42 / Chapter 2.1.3 --- "Cell culture medium, buffers and other reagents" --- p.43 / Chapter 2.1.4 --- Radioisotope and scintillation fluid --- p.47 / Chapter 2.1.5 --- Reagents and buffers for flow cytometry --- p.48 / Chapter 2.1.6 --- Antibodies for flow cytometry --- p.50 / Chapter 2.1.7 --- Recombinant cytokines --- p.51 / Chapter 2.1.8 --- Reagents for DNA extraction --- p.52 / Chapter 2.1.9 --- Reagents for total RNA isolation --- p.53 / Chapter 2.1.10 --- Reagents and buffers for RT-PCR --- p.54 / Chapter 2.1.11 --- Reagents and buffers for gel electrophoresis --- p.58 / Chapter 2.1.12 --- Reagents and buffers for Western blot analysis --- p.59 / Chapter 2.1.13 --- Reagents for measuring caspase activity --- p.64 / Chapter 2.2 --- Methods --- p.67 / Chapter 2.2.1 --- Culture of the leukemia cell lines --- p.67 / Chapter 2.2.2 --- Isolation and preparation of normal hematopoietic cells --- p.67 / Chapter 2.2.3 --- [3 H]-TdR proliferation assay --- p.68 / Chapter 2.2.4 --- Determination of cell viability --- p.69 / Chapter 2.2.5 --- Assay for anti-leukemic activity in vivo --- p.70 / Chapter 2.2.6 --- Assessment of differentiation-associated characteristics --- p.71 / Chapter 2.2.7 --- Assays for apoptosis --- p.73 / Chapter 2.2.8 --- Cell cycle analysis (DNA content evaluation) --- p.75 / Chapter 2.2.9 --- Gene expression study --- p.75 / Chapter 2.2.10 --- Protein expression study --- p.79 / Chapter 2.2.11 --- Measurement of caspase activity --- p.82 / Chapter 2.2.12 --- Statistical analysis 一 --- p.83 / Chapter CHAPTER 3: --- STUDIES ON THE ANTI-PROLIFERATIVE EFFECT OF TOPOISOMERASE-TARGETING AGENTS ON LEUKEMIA CELLS / Chapter 3.1 --- Introduction --- p.84 / Chapter 3.2 --- Results --- p.86 / Chapter 3.2.1 --- The anti-proliferative effect of topoisomerase-targeting agents on human and murine leukemia cells in vitro --- p.86 / Chapter 3.2.2 --- Effect of 10-hydroxy-camptothecin and etoposide phosphate on the clonogenicity of the murine myeloid leukemia WEHI-3B JCS cells in vitro --- p.105 / Chapter 3.2.3 --- Effects of 10-hydroxy-camptothecin and etoposide phosphate on the tumorigenicity and proliferation of the murine myeloid leukemia WEHI-3B JCS cells invivo --- p.106 / Chapter 3.2.4 --- Cytotoxic effect of 10-hydroxy-camptothecin and etoposide phosphate on normal hematopoietic cells and WEHI-3B JCS cells in vitro --- p.109 / Chapter 3.2.5 --- Effect of 10-hydroxy-camptothecin and etoposide phosphate on the cell cycle kinetics of WEHI-3B JCS cells --- p.114 / Chapter 3.2.6 --- Effect of 10-hydroxy-camptothecin and etoposide phosphate on the expression of cell cycle-regulatory genes in the murine myeloid leukemia WEHI-3B JCS cells --- p.116 / Chapter 3.2.7 --- Combination effect of 10-hydroxy-camptothecin or etoposide phosphate with cytokines on the proliferation of the murine myeloid leukemia WEHI-3B JCS cells --- p.123 / Chapter 3.3 --- Discussion --- p.127 / Chapter CHAPTER 4: --- STUDIES ON THE DIFFERENTIATION-INDUCING EFFECT OF 10-HYDROXY-CAMPTOTHECIN AND ETOPOSIDE PHOSPHATE ON THE MURINE MYELOID LEUKEMIA WEHI-3B JCS CELLS / Chapter 4.1 --- Introduction --- p.132 / Chapter 4.2 --- Results --- p.134 / Chapter 4.2.1 --- Morphological changes in the murine myeloid leukemia WEHI-3B JCS cells treated with 10-hydroxy-camptothecin and etoposide phosphate --- p.134 / Chapter 4.2.2 --- Effect of 10-hydroxy-camptothecin and etoposide phosphate on the size and granularity of the murine myeloid leukemia WEHI-3B JCS cells --- p.138 / Chapter 4.2.3 --- Effect of 10-hydroxy-camptothecin and etoposide phosphate on the plastic adhering property of the murine myeloid leukemia WEHI-3B JCS cells --- p.140 / Chapter 4.2.4 --- Effect of 10-hydroxy-camptothecin and etoposide phosphate on the NBT-reducing activity of the murine myeloid leukemia WEHI-3B JCS cells --- p.142 / Chapter 4.2.5 --- Surface antigen immunophenotyping of the murine myeloid leukemia WEHI-3B JCS cells treated with 10-hydroxy- camptothecin and etoposide phosphate --- p.145 / Chapter 4.2.6 --- Induction of non-specific esterase activity in the murine myeloid leukemia WEHI-3B JCS cells by 10-hydroxy- camptothecin and etoposide phosphate --- p.152 / Chapter 4.3 --- Discussion --- p.154 / Chapter CHAPTER 5: --- STUDIES ON THE APOPTOSIS-INDUCING EFFECT OF 10-HYDROXY-CAMPTOTHECIN AND ETOPOSIDE PHOSPHATE ON THE MURINE MYELOID LEUKEMIA WEHI-3B JCS CELLS / Chapter 5.1 --- Introduction --- p.157 / Chapter 5.2 --- Results --- p.160 / Chapter 5.2.1 --- Induction of nuclear disintegration in the murine myeloid leukemia WEHI-3B JCS cells by 10-hydroxy-camptothecin and etoposide phosphate --- p.160 / Chapter 5.2.2 --- Induction of DNA fragmentation in the murine myeloid leukemia WEHI-3B JCS cells by 10-hydroxy-camptothecin and etoposide phosphate --- p.162 / Chapter 5.2.3 --- Induction of phosphatidylserine translocation in murine myeloid leukemia WEHI-3B JCS cells by 10-hydroxy- camptothecin and etoposide phosphate --- p.167 / Chapter 5.2.4 --- Effect of 10-hydroxy-camptothecin and etoposide phosphate on the expression of apoptosis-regulatory genes in the murine myeloid leukemia WEHI-3B JCS cells --- p.171 / Chapter 5.2.5 --- Effect of 10-hydroxy-camptothecin and etoposide phosphate on the expression of apoptosis-regulatory proteins in the murine myeloid leukemia WEHI-3B JCS cells --- p.177 / Chapter 5.2.6 --- Induction of mitochondrial membrane depolarization in the murine myeloid leukemia WEHI-3B JCS cells by 10-hydroxy- camptothecin and etoposide phosphate --- p.179 / Chapter 5.2.7 --- Effect of 10-hydroxy-camptothecin and etoposide phosphate on caspase activity in the murine myeloid leukemia WEHI-3B JCS cells --- p.181 / Chapter 5.2.8 --- Effect of 10-hydroxy-camptothecin and etoposide phosphate intracellular Ca2+ level in the murine myeloid leukemia WEHI-3B JCS cells --- p.186 / Chapter 5.3 --- Discussion --- p.189 / Chapter CHAPTER 6: --- CONCLUSIONS AND FUTURE PERSPECTIVES --- p.192 / REFERENCES --- p.199

Identiferoai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_324471
Date January 2003
ContributorsChan, Wai Sing., Chinese University of Hong Kong Graduate School. Division of Biochemistry.
Source SetsThe Chinese University of Hong Kong
LanguageEnglish, Chinese
Detected LanguageEnglish
TypeText, bibliography
Formatprint, xvii, 210 leaves : ill. ; 30 cm.
RightsUse 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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