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Studies on the anti-tumor activities of conjugated linolenic acid on human neuroblastoma cells.

Ho, Lai Ying. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2009. / Includes bibliographical references (leaves 213-238). / Abstract also in Chinese. / Abstract --- p.i / Abstract in Chinese (摘要) --- p.iv / Acknowledgements --- p.vii / List of abbreviations --- p.ix / Table of contents --- p.xiv / Chapter Chapter One: --- General Introduction_ --- p.1 / Chapter 1.1 --- Neuroblastoma --- p.2 / Chapter 1.1.1 --- An overview on neuroblastoma --- p.2 / Chapter 1.1.2 --- Classification of neuroblastoma --- p.4 / Chapter 1.1.3 --- Epidemiology of neuroblastoma --- p.7 / Chapter 1.1.4 --- Clinical manifestation of neuroblastoma --- p.10 / Chapter 1.1.5 --- Diagnosis of neuroblastoma --- p.10 / Chapter 1.1.6 --- Conventional therapy of neuroblastoma --- p.12 / Chapter 1.1.7 --- Novel treatments of neuroblastoma --- p.14 / Chapter 1.2 --- Conjugated linolenic acid (CLN) --- p.16 / Chapter 1.2.1 --- An overview of polyunsaturated fatty acids and conjugated fatty acids --- p.16 / Chapter 1.2.2 --- Chemical structure and physical properties of CLNs --- p.18 / Chapter 1.2.3 --- Natural occurrence of CLNs --- p.21 / Chapter 1.2.4 --- Synthesis of CLNs --- p.22 / Chapter 1.2.5 --- Metabolism and pharmacokinetics of CLNs --- p.24 / Chapter 1.2.6 --- Major biological and pharmacological activities of CLNs --- p.25 / Chapter 1.2.6.1 --- Hypolipidemic and anti-obese effects --- p.25 / Chapter 1.2.6.2 --- Anti-cancer effects --- p.27 / Chapter 1.2.6.2.1 --- Anti-proliferation --- p.27 / Chapter 1.2.6.2.2 --- Chemoprevention --- p.28 / Chapter 1.2.6.2.3 --- Apoptosis-inducing --- p.28 / Chapter 1.3 --- Aims and scope of the study --- p.31 / Chapter Chapter Two: --- Materials and Methods_ --- p.34 / Chapter 2.1 --- Materials --- p.35 / Chapter 2.1.1 --- Animals --- p.35 / Chapter 2.1.2 --- Cell lines --- p.35 / Chapter 2.1.3 --- "Cell culture medium, buffers and other reagents" --- p.37 / Chapter 2.1.4 --- Reagents for DNA extraction --- p.46 / Chapter 2.1.5 --- Reagents for gel electrophoresis of nucleic acids --- p.47 / Chapter 2.1.6 --- Reagents and buffers for flow cytometry --- p.49 / Chapter 2.1.7 --- Reagents and buffers for measuring caspase activity --- p.50 / Chapter 2.1.8 --- Reagents for Hoechst staining --- p.53 / Chapter 2.1.9 --- Reagents and buffers for RNA extraction --- p.53 / Chapter 2.1.10 --- Reagents and buffers for DNA digestion --- p.54 / Chapter 2.1.11 --- Reagents and buffers for reverse transcription --- p.55 / Chapter 2.1.12 --- Reagents for real-time polymerase chain reaction --- p.57 / Chapter 2.1.13 --- Reagents and buffers for Western blotting --- p.59 / Chapter 2.2 --- Methods --- p.64 / Chapter 2.2.1 --- Culture of cell lines --- p.64 / Chapter 2.2.2 --- Preparation of NIH-3T3 conditioned medium --- p.65 / Chapter 2.2.3 --- Determination of cell viability --- p.65 / Chapter 2.2.4 --- Determination of cell proliferation by tritiated thymidine incorporation assay --- p.66 / Chapter 2.2.5 --- Cytotoxicity test of CLNs on murine peritoneal macrophages --- p.67 / Chapter 2.2.6 --- Cytotoxicity test of CLNs on murine bone marrow cells --- p.68 / Chapter 2.2.7 --- Cytotoxicity test of CLNs on murine splenocytes --- p.68 / Chapter 2.2.8 --- Cytotoxicity tests of CLNs on human peripheral blood mononuclear cells --- p.69 / Chapter 2.2.9 --- Carboxyfluorescein diacetate succinimidyl ester (CFSE) staining analyzed by flow cytometry --- p.69 / Chapter 2.2.10 --- Determination of colony forming ability --- p.70 / Chapter 2.2.11 --- Determination of cell invasiveness --- p.70 / Chapter 2.2.12 --- In vivo tumorigenicity assay --- p.71 / Chapter 2.2.13 --- Analysis of cell cycle profile/ DNA content by flow cytometry --- p.72 / Chapter 2.1.14 --- Measurements of apoptosis --- p.72 / Chapter 2.1.15 --- Measurements of differentiation --- p.77 / Chapter 2.1.16 --- Gene expression study --- p.78 / Chapter 2.2.17 --- Protein expression study --- p.81 / Chapter 2.2.18 --- Statistical Analysis --- p.84 / Chapter Chapter Three: --- Anti-proliferative Effect of CLN Isomers on Human Neuroblastoma cells --- p.86 / Chapter 3.1 --- Introduction --- p.87 / Chapter 3.2 --- Results --- p.89 / Chapter 3.2.1 --- Anti-proliferative effect of CLN isomers on various human neuroblastoma cell lines in vitro --- p.89 / Chapter 3.2.2 --- Direct cytotoxic effect of jacaric acid on neuroblastoma LA-N-1 cells in vitro --- p.100 / Chapter 3.2.3 --- Cytotoxicity of jacaric acid on primary murine cells and human normal cell lines --- p.103 / Chapter 3.2.4 --- Kinetic and reversibility studies of the anti-proliferative effect of jacaric acid on LA-N-1 cells --- p.106 / Chapter 3.2.5 --- Synergistic anti-proliferative effect of jacaric acid with daidzein and retinoic acid on LA-N-1 cells in vitro --- p.110 / Chapter 3.2.6 --- Modulatory effect of jacaric acid on the number of cell division in LA-N-1 cells --- p.113 / Chapter 3.2.7 --- Effect of jacaric acid on the cell cycle profile of LA-N-1 cells --- p.115 / Chapter 3.2.8 --- Effect of jacaric acid on the invasiveness of LA-N-1 cells --- p.118 / Chapter 3.2.9 --- Effect of jacaric acid on the colony forming ability of LA-N-1 cells in soft agar --- p.120 / Chapter 3.2.10 --- Effect of jacaric acid on the in vivo tumorigenicity of the LA-N-1 cells --- p.122 / Chapter 3.3 --- Discussion --- p.124 / Chapter Chapter Four: --- Apoptosis- and Differentiation-inducing Effects of Jacaric Acid on Human Neuroblastoma Cells --- p.133 / Chapter 4.1 --- Introduction --- p.134 / Chapter 4.2 --- Results --- p.138 / Chapter 4.2.1 --- Induction of DNA fragmentation and apoptotic ultrastructural changes in LA-N-1 cells by jacaric acid --- p.138 / Chapter 4.2.2 --- Induction of phosphatidylserine externalization by jacaric acid in human neuroblastoma cells as detected by Annexin V-GFP/ PI dual staining --- p.142 / Chapter 4.2.3 --- Effect of jacaric acid on the mitochondrial membrane potential in human neuroblastoma cells --- p.146 / Chapter 4.2.4 --- Effect of jacaric acid on the caspase-3 activity in LA-N-1 cells --- p.150 / Chapter 4.2.5 --- Effect of jacaric acid on the reactive oxygen species (ROS) generation in human neuroblastoma cells --- p.153 / Chapter 4.2.6 --- Morphological changes induced by jacaric acid in SH-SY5Y cells --- p.158 / Chapter 4.3 --- Discussion --- p.162 / Chapter Chapter Five: --- Mechanistic Studies of Anti-proliferative Effect of Jacaric Acid on Human Neuroblastoma Cells --- p.171 / Chapter 5.1 --- Introduction --- p.172 / Chapter 5.2 --- Results --- p.178 / Chapter 5.2.1 --- Effect of antioxidant a-tocopherol on the anti-proliferative effect of jacaric acid on LA-N-1 cells --- p.178 / Chapter 5.2.2 --- Effect of caspase inhibitors on the anti-proliferative effect of jacaric acid on LA-N-1 cells --- p.180 / Chapter 5.2.3 --- Jacaric acid modulated the mRNA expression of N-myc and other related transcription factors in LA-N-1 cells --- p.182 / Chapter 5.2.4 --- Jacaric acid modulated the protein expression of N-myc --- p.186 / Chapter 5.2.5 --- Jacaric acid modulated the mRNA expression of apoptosis-associated genes --- p.188 / Chapter 5.3 --- Discussion --- p.191 / Chapter Chapter Six: --- Conclusions and Future Perspectives --- p.202 / References --- p.212

Identiferoai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_326751
Date January 2009
ContributorsHo, Lai Ying., Chinese University of Hong Kong Graduate School. Division of Life Sciences.
Source SetsThe Chinese University of Hong Kong
LanguageEnglish, Chinese
Detected LanguageEnglish
TypeText, bibliography
Formatprint, xviii, 238 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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