Gho Wai-Man. / Thesis submitted in: August 2006. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2007. / Includes bibliographical references (leaves 114-123). / Abstracts in English and Chinese. / ABSTRACT --- p.I / 摘要 --- p.IV / ACKNOWLEDGEMENT --- p.VI / TABLE OF CONTENT --- p.VII / LIST OF TABLES --- p.XII / LIST OF FIGURES --- p.XIII / ABBREVIATIONS --- p.XVI / Chapter CHAPTER 1 --- INTRODUCTION --- p.1 / Chapter 1.1 --- Introduction --- p.2 / Chapter 1.2 --- Epidemiology --- p.2 / Chapter 1.3 --- Etiological factors --- p.6 / Chapter 1.3.1 --- Viral Hepatitis Infection --- p.6 / Chapter 1.3.1.1 --- Hepatitis B Virus (HBV) --- p.7 / Chapter 1.3.1.2 --- Hepatitis C Virus (HCV) --- p.9 / Chapter 1.3.2 --- Aflatoxin Exposure --- p.10 / Chapter 1.3.3 --- Alcohol Abuse --- p.11 / Chapter 1.3.4 --- Liver Cirrhosis --- p.12 / Chapter 1.4 --- Genetic alterations in hcc --- p.16 / Chapter 1.4.1 --- Chromosomal Gain --- p.16 / Chapter 1.4.2 --- Chromosomal Loss --- p.17 / Chapter 1.5 --- Discovery of common activating transcription factor 5 (atf5) down-regulations in hcc --- p.19 / Chapter 1.5.1 --- Chromosome 19 Aberration in HCC --- p.19 / Chapter 1.5.2 --- Discovery of High Frequency of　ATF5 Down-regulations --- p.19 / Chapter 1.5.3 --- Activating Transcription Factor Family --- p.20 / Chapter 1.6 --- Aim of thesis --- p.28 / Chapter CHAPTER 2 --- MATERIALS AND METHODS --- p.29 / Chapter 2.1 --- Materials --- p.30 / Chapter 2.1.1 --- Chemicals --- p.30 / Chapter 2.1.2 --- Buffers --- p.31 / Chapter 2.1.3 --- Cell culture --- p.31 / Chapter 2.1.4 --- Nucleic acids --- p.32 / Chapter 2.1.5 --- Enzymes --- p.32 / Chapter 2.1.6 --- Equipment --- p.32 / Chapter 2.1.7 --- Kits --- p.33 / Chapter 2.1.8 --- Software and Web Resource --- p.33 / Chapter 2.2 --- Dna extraction --- p.34 / Chapter 2.2.1 --- Cell Lines --- p.34 / Chapter 2.2.2 --- Primary HCC --- p.34 / Chapter 2.2.3 --- Lymphocytic DNA --- p.35 / Chapter 2.3 --- Rna extraction --- p.36 / Chapter 2.4 --- Dna sequencing --- p.38 / Chapter 2.4.1 --- Polymerase Chain Reaction (PCR) --- p.38 / Chapter 2.4.2 --- Cycle Sequencing --- p.39 / Chapter 2.5 --- Dual-labeled fluirescence in situ hybridization (fish) --- p.41 / Chapter 2.5.1 --- FISH Probe Preparation --- p.41 / Chapter 2.5.1.1 --- Preparation of Human Bacterial Artificial Chromosome (BAC) --- p.41 / Chapter 2.5.1.2 --- Nick Translation --- p.41 / Chapter 2.5.2 --- FISH --- p.42 / Chapter 2.6 --- 5-aza-2'-deoxycytidine & trichostatin a treatment on cell lines --- p.43 / Chapter 2.7 --- Bisulfite modificaiton of dna --- p.43 / Chapter 2.8 --- Methylation-specific pcr (msp) --- p.44 / Chapter 2.9 --- Bisulfite dna sequencing --- p.44 / Chapter 2.10 --- Quantitative reverse transcription pcr (qrt-pcr) --- p.46 / Chapter 2.11 --- In-vitro and in-vivo functinal examination --- p.49 / Chapter 2.11.1 --- ATF5 Transfection --- p.49 / Chapter 2.11.2 --- Cell Growth Assay --- p.50 / Chapter 2.11.3 --- Xenograft Development --- p.51 / Chapter 2.12 --- codelink expression microarray --- p.51 / Chapter 2.13 --- Statistical analysis --- p.53 / Chapter CHAPTER 3 --- INACTIVATION OF MECHANISMS UNDERLYING ATF5 DOWN-REGULATION --- p.54 / Chapter 3.1 --- Introduction --- p.55 / Chapter 3.2 --- Materials and methods --- p.58 / Chapter 3.2.1 --- Cell Lines --- p.58 / Chapter 3.2.2 --- Mutational Analysis --- p.58 / Chapter 3.2.3 --- Copy Number Loss --- p.59 / Chapter 3.2.4 --- Epigenetic Control --- p.59 / Chapter 3.3 --- Results --- p.67 / Chapter 3.3.1 --- Sequencing Analysis of A TF5 Gene --- p.67 / Chapter 3.3.2 --- FISH Analysis of ATF5 Copy Number --- p.73 / Chapter 3.3.3 --- Epigenetic Control of A TF5 Expression --- p.73 / Chapter 3.4 --- Discussion --- p.82 / Chapter CHAPTER 4 --- FUNCTIONAL EXAMINATION AND INVESTIGATION OF DOWNSTREAM TARGETS MODULATED BY ATF5 --- p.85 / Chapter 4.1 --- Introduction --- p.86 / Chapter 4.2 --- Materials and methods --- p.88 / Chapter 4.2.1 --- Cell Lines --- p.88 / Chapter 4.2.2 --- Plasmids and Transfection --- p.88 / Chapter 4.2.3 --- Cell Growth Assay --- p.88 / Chapter 4.2.4 --- Xenograft Development --- p.88 / Chapter 4.2.5 --- CodeLink Expression Microarray --- p.89 / Chapter 4.2.6 --- Quantitative RT-PCR --- p.90 / Chapter 4.2.7 --- Statistical analysis --- p.90 / Chapter 4.3 --- Results --- p.91 / Chapter 4.3.1 --- Cell Proliferation --- p.91 / Chapter 4.3.1.1 --- In-Vitro Examination --- p.91 / Chapter 4.3.1.2 --- In-Vivo Examination --- p.91 / Chapter 4.3.2 --- Microarray A nalysis --- p.91 / Chapter 4.3.3 --- Correlation of A TF5 with Id-1 Expression --- p.103 / Chapter 4.4 --- Discussion --- p.106 / Chapter CHAPTER 5 --- PROPOSED FUTURE INVESTIGATIONS --- p.110 / Chapter 5.1 --- inactivation mechanisms of atf5 gene --- p.111 / Chapter 5.2 --- Molecular pathways modulated by atf5 --- p.112 / Chapter CHAPTER 6 --- REFERENCES --- p.114

by Sung Ying-Man, Mandy. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2003. / Includes bibliographical references (leaves 112-125). / Abstracts in English and Chinese. / Acknowledgments --- p.I / List of Abbreviations --- p.II / Abstract --- p.IV / 摘要 --- p.VII / Chapter Chapter 1 --- Introduction / Chapter 1.1 --- Epidemiology --- p.1 / Chapter 1.2 --- Aetiological factors --- p.4 / Chapter 1.2.1 --- Hepatitis B infection --- p.4 / Chapter 1.2.2 --- Aflatoxin exposure --- p.5 / Chapter 1.2.3 --- Alcohol consumption --- p.5 / Chapter 1.2.4 --- Genetic risk factors --- p.6 / Chapter 1.3 --- Aims of the study --- p.7 / Chapter Chapter 2 --- Polymorphisms of candidate genes in Interleukin- signalling pathway in HCC --- p.6 / Chapter 2.1 --- Introduction --- p.9 / Chapter 2.2 --- Materials and Methods --- p.15 / Samples and Genomic DNA isolation --- p.15 / PCR-PFLP --- p.15 / dHPLC --- p.16 / Direct Sequencing --- p.17 / Stattistical Analysis --- p.17 / Chapter 2.3 --- Results --- p.25 / Chapter 2.3.1 --- Known IL6 polymorphisms --- p.25 / Chapter 2.3.2 --- IL-6R and gp130 polymorphisms --- p.28 / Chapter 2.3.3 --- Stat-3 polymorphisms --- p.29 / Chapter 2.3.4 --- SOCS-1 polymophisms --- p.32 / Chapter 2.3.5 --- Mutation screening of IL-6 gene --- p.34 / Chapter 2.3.6 --- Mutation screening of SOCS-1 gene --- p.39 / Chapter 2.4 --- Discussion --- p.40 / Chapter 2.4.1 --- Interleukin-6 --- p.40 / Chapter 2.4.2 --- Gp130 and IL6-R --- p.43 / Chapter 2.4.3 --- STAT-3 --- p.44 / Chapter 2.4.4 --- SOCS-l --- p.46 / Chapter Chapter 3 --- Methylation status of SOCS-1 gene in Chinese HCC patients / Chapter 3.1 --- Introduction --- p.49 / Chapter 3.2 --- Methods and Materials --- p.53 / Tissue Sampling --- p.53 / Methylation specific PCR (MSP) --- p.53 / Chapter 3.3 --- Results --- p.55 / Chapter 3.4 --- Discussion --- p.56 / Chapter Chapter 4 --- Polymorphisms of enzyme encoding genesin steroidogenesis in Chinese / Chapter 4.1 --- Introduction --- p.59 / Chapter 4.1.1 --- Steroid 5a reductases (SRD5A) --- p.62 / Chapter 4.1.1a --- Steroid 5a reductase type II (SRD5A2) --- p.63 / Chapter 4.1.1b --- Steroid 5a reductase type I (SRD5A1) --- p.65 / Chapter 4.1.2 --- Cytochrome P450al7 (CYP17) --- p.67 / Chapter 4.1.3 --- "Cytochrome P450, family 1，subfamily A polypeptide1 (CYP1A1)" --- p.69 / Chapter 4.1.4 --- "Cytochrome P450, subfamily IIIA (niphedipine oxidase) polypeptide 4 (CYP3A4)" --- p.71 / Chapter 4.2 --- Materials and Methods --- p.74 / Samples and Genomic DNA isolation --- p.74 / PCR-PFLP --- p.74 / Direct Sequencing --- p.74 / Statistical Analysis --- p.74 / Chapter 4.3 --- Results --- p.77 / Chapter 4.3.1 --- SRD5A2 --- p.77 / Chapter 4.3.2 --- Linkage Disequilibrium in SRD5A2 gene --- p.83 / Chapter 4.3.2 --- SRD5A1 --- p.84 / Chapter 4.3.3 --- CYP17 --- p.87 / Chapter 4.3.4 --- CYP1A1 --- p.89 / Chapter 4.3.5 --- CYP3A4 --- p.92 / Chapter 4.3.6 --- Logistic regression --- p.95 / Chapter 4.4 --- Discussion --- p.96 / Chapter 4.4.1 --- SRD5A2 --- p.96 / Chapter 4.4.2 --- SRD5A1 --- p.99 / Chapter 4.4.3 --- CYP17 --- p.101 / Chapter 4.4.4 --- CYP1A1 --- p.103 / Chapter 4.4.5 --- CYP3A4 --- p.106 / Chapter 4.4.6 --- Logistic Regression --- p.107 / Chapter Chapter 5 --- Conclusions and Future Prospect --- p.108 / Chapter 5.1 --- Conclusions --- p.108 / Chapter 5.2 --- Future works and prospect --- p.111 / References --- p.113