Anti-proliferative and differentiation-inducing effects of glycyrrhizin and 18[beta]-glycyrrhetinic acid on neuroblastoma cells in vitro.

• Other Authors: Lee, Kin-wah.
• Format: Others
• Language: English; Chinese
• Published: 2003
• Subjects: Neuroblastoma; Licorice (Plant); Glycyrrhiza
• Online Access: http://library.cuhk.edu.hk/record=b5896083; http://repository.lib.cuhk.edu.hk/en/item/cuhk-324417

Lee Kin-wah. === Thesis (M.Phil.)--Chinese University of Hong Kong, 2003. === Includes bibliographical references (leaves 187-203). === Abstracts in English and Chinese. === ACKNOWLEDGEMENTS --- p.i === ABBREVIATIONS --- p.ii === ABSTRACT --- p.vi === CHINESE ABSTRACT --- p.ix === TABLE OF CONTENTS --- p.xii === Chapter CHAPTER 1: --- GENERAL INTRODUCTION --- p.1 === Chapter 1.1 --- Neuroblastoma-An Overview --- p.1 === Chapter 1.1.1 --- Epidemiology of Neuroblastoma --- p.1 === Chapter 1.1.2 --- Clinical Presentations of Neuroblastoma --- p.2 === Chapter 1.1.3 --- Diagnosis of Neuroblastoma --- p.3 === Chapter 1.1.4 --- Staging of Neuroblastoma --- p.5 === Chapter 1.1.5 --- Prognostic Considerations of Neuroblastoma --- p.6 === Chapter 1.1.5.1 --- Age --- p.6 === Chapter 1.1.5.2 --- Primary Tumor Site --- p.6 === Chapter 1.1.5.3 --- Tumor Histopathology --- p.7 === Chapter 1.1.5.4 --- Serum Markers --- p.9 === Chapter 1.1.5.5 --- Genetic Markers --- p.9 === Chapter 1.1.6 --- Standard Treatment Modalities for Neuroblastoma --- p.11 === Chapter 1.1.6.1 --- Surgery --- p.11 === Chapter 1.1.6.2 --- Chemotherapy --- p.12 === Chapter 1.1.6.3 --- Radiotherapy --- p.13 === Chapter 1.1.7 --- Differentiation of Neuroblastoma In Vivo and In Vitro --- p.14 === Chapter 1.1.8 --- Differentiation Therapy of Neuroblastoma --- p.16 === Chapter 1.2 --- Glycyrrhizin ´ؤ the Major Active Component of Licorice --- p.17 === Chapter 1.2.1 --- Chemistry of Licorice --- p.17 === Chapter 1.2.2 --- Metabolism of Glycyrrhizin --- p.21 === Chapter 1.2.3 --- Pharmacological Effects of Glycyrrhizin and 18β-Glycyrrhetinic Acid --- p.22 === Chapter 1.2.3.1 --- Anti-inflammatory Effect --- p.22 === Chapter 1.2.3.2 --- Hepatoprotective Effect --- p.23 === Chapter 1.2.3.3 --- Anti-carcinogenic and Anti-tumor Effects --- p.25 === Chapter 1.2.3.4 --- Anti-viral Effect --- p.27 === Chapter 1.2.3.5 --- Immunomodulatory Effect --- p.29 === Chapter 1.2.3.6 --- Mineralocorticoid Effect --- p.30 === Chapter 1.2.4 --- Pharmacokinetics of Glycyrrhizin and Glycyrrhetinic Acid --- p.32 === Chapter 1.2.5 --- Health Hazards of Glycyrrhizin and Glycyrrhetinic Acid --- p.33 === Chapter 1.3 --- Aims and Scopes of This Study --- p.36 === Chapter CHAPTER 2: --- MATERIALS AND METHODS --- p.39 === Chapter 2.1 --- Materials --- p.39 === Chapter 2.1.1 --- Cell Lines --- p.39 === Chapter 2.1.2 --- "Cell Culture Media, Buffers and Other Reagents" --- p.39 === Chapter 2.1.3 --- Drugs and Chemicals --- p.43 === Chapter 2.1.4 --- Reagents for 3H-Thymidine Incorporation Assay --- p.44 === Chapter 2.1.5 --- Reagents for Neutral Red Assay --- p.45 === Chapter 2.1.6 --- Reagents for Clonogenic Assay --- p.45 === Chapter 2.1.7 --- Reagents and Buffers for Immunocytochemistry --- p.46 === Chapter 2.1.8 --- Reagents for DNA Extraction --- p.48 === Chapter 2.1.9 --- Reagent for DNA Staining --- p.49 === Chapter 2.1.10 --- Reagents and Buffers for Flow Cytometry --- p.49 === Chapter 2.1.11 --- Reagents for Total RNA Isolation --- p.50 === Chapter 2.1.12 --- Reagents and Buffers for RT-PCR --- p.50 === Chapter 2.1.13 --- Reagents and Buffers for Gel Electrophoresis --- p.55 === Chapter 2.1.14 --- Reagents and Buffers for Western Blot Analysis --- p.56 === Chapter 2.2 --- Methods --- p.62 === Chapter 2.2.1 --- Cell Culture Methodology --- p.62 === Chapter 2.2.2 --- Determination of Cell Proliferation --- p.62 === Chapter 2.2.3 --- Determination of Cell Viability by Trypan Blue Exclusion Test --- p.64 === Chapter 2.2.4 --- Limiting Dilution Assay --- p.65 === Chapter 2.2.5 --- Clonogenic Assay --- p.65 === Chapter 2.2.6 --- Measurement of Apoptosis by DNA Fragmentation Analysis --- p.66 === Chapter 2.2.7 --- Assessment of Apoptosis by Hoechst 33342 Staining --- p.67 === Chapter 2.2.8 --- Cell Morphological Study --- p.67 === Chapter 2.2.9 --- Immunocytochemistry --- p.68 === Chapter 2.2.10 --- Flow Cytometric Analysis of Cell Cycle Profile --- p.69 === Chapter 2.2.11 --- Gene Expression Study --- p.70 === Chapter 2.2.12 --- Protein Expression Study --- p.73 === Chapter 2.2.13 --- Statistical Analysis --- p.76 === Chapter CHAPTER 3: --- ANTI-PROLIFERATIVE EFFECTS OF GLYCYRRHIZIN AND 18β-GLYCYRRHETINIC ACID ON NEUROBLASTOMA CELLS --- p.77 === Chapter 3.1 --- Introduction --- p.77 === Chapter 3.2 --- Results --- p.79 === Chapter 3.2.1 --- Differential Anti-proliferative Effect of Glycyrrhizin and 18β- Glycyrrhetinic Acid on Various Neuroblastoma Cell Lines In Vitro --- p.79 === Chapter 3.2.2 --- Effect of 18P-Glycyrrhetinic Acid on the Clonogenicity of the Murine Neuroblastoma BU-1 Cells In Vitro --- p.91 === Chapter 3.2.3 --- Kinetic and Reversibility Studies of the Anti-proliferative Effect of Glycyrrhizin and 18β-Glycyrrhetinic Acid on the Neuroblastoma BU-1 Cells --- p.93 === Chapter 3.2.4 --- Cytotoxic Effect of Glycyrrhizin and 18β-Glycyrrhetinic Acid on the Neuroblastoma BU-1 Cells In Vitro --- p.100 === Chapter 3.2.5 --- Inability of Glycyrrhizin and 18β-Glycyrrhetinic Acid to Induce DNA Fragmentation in the Neuroblastoma BU-1 Cells --- p.102 === Chapter 3.3 --- Discussion --- p.107 === Chapter CHAPTER 4: --- DIFFERENTIATION-INDUCING EFFECTS OF GLYCYRRHIZIN AND 18β-GLYCYRRHETINIC ACID ON NEUROBLASTOMA CELLS --- p.112 === Chapter 4.1 --- Introduction --- p.112 === Chapter 4.2 --- Results --- p.114 === Chapter 4.2.1 --- Morphological Changes in Glycyrrhizin and 18β-Glycyrrhetinic Acid-treated Neuroblastoma BU-1 Cells --- p.114 === Chapter 4.2.2 --- Immunocytochemistry of Glycyrrhizin and 18β-Glycyrrhetinic Acid-treated Neuroblastoma BU-1 Cells --- p.118 === Chapter 4.2.3 --- Effect of 18β-Glycyrrhetinic Acid on the Expression of Proto-oncogenes in Neuroblastoma BU-1 Cells --- p.124 === Chapter 4.2.4 --- Effect of 18β-Glycyrrhetinic Acid on the Expression of Differentiation-Related Genes in Neuroblastoma BU-1 Cells --- p.127 === Chapter 4.3 --- Discussion --- p.130 === Chapter CHAPTER 5: --- MECHANISTIC STUDIES ON THE ANTI-PROLIFERATIVE AND DIFFERENTIATION-INDUCING EFFECTS OF GLYCYRRHIZIN AND 18β-GLYCYRRHETINIC ACID --- p.136 === Chapter 5.1 --- Introduction --- p.136 === Chapter 5.2 --- Results --- p.139 === Chapter 5.2.1 --- Effects of Glycyrrhizin and 18β-Glycyrrhetinic Acid on the Cell Cycle Kinetics of Neuroblastoma BU-1 Cells In Vitro --- p.139 === Chapter 5.2.2 --- Modulatory Effects of 18β-Glycyrrhetinic Acid on the Expression of Cell Cycle Regulatory Genes and Proteins --- p.145 === Chapter 5.2.3 --- "Combined Effects of Glycyrrhizin, 18β-Glycyrrhetinic Acid and All-Trans Retinoic Acid on the Proliferation of Neuroblastoma BU-1 Cells In Vitro" --- p.149 === Chapter 5.2.4 --- "Combined Effects of Glycyrrhizin, 18β-Glycyrrhetinic Acid and All-Trans Retinoic Acid on the Differentiation of Neuroblastoma BU-1 Cells In Vitro" --- p.153 === Chapter 5.2.5 --- Modulatory Effect of 18β-Glycyrrhetinic Acid on the Expression of PKC Isoforms in Neuroblastoma BU-1 Cells --- p.156 === Chapter 5.2.6 --- The Possible Involvement of Protein Kinase C in the Anti-proliferative and Differentiation-Inducing Effects of 18β-Glycyrrhetinic Acid on the Neuroblastoma BU-1 Cells --- p.158 === Chapter 5.2.7 --- The Possible Involvement of Protein Kinase A in the Anti-proliferative and Differentiation-Inducing Effects of 18β-Glycyrrhetinic Acid on the Neuroblastoma BU-1 Cells --- p.165 === Chapter 5.3 --- Discussion --- p.173 === Chapter CHAPTER 6: --- CONCLUSIONS AND FUTURE PERSPECTIVES --- p.182 === REFERENCES --- p.187