Title Page
감사의 글
Contents
ABBREVIATIONS 13
CHAPTER I. INTRODUCTION 14
1-1. THE LIVER 15
1-2. FUNCTION OF THE LIVER 15
1-3. VIROLOGICAL MARKERS 16
1-4. LIVER DISEASES 16
1-4-1. ACUTE AND CHRONIC VIRAL HEPATITIS 16
1-4-2. HBV CLEARANCE 17
1-4-3. CHRONIC HEPATITIS B 17
1-4-4. CHRONIC HEPATITIS C 18
1-4-5. ALCOHOL INDUCED LIVER DISEASES 18
1-4-6. DRUG INDUCED LIVER DISEASES 19
1-4-7. CIRRHOSIS AND LIVER FIBROSIS 19
1-4-8. HEPATOCELLULAR CARCINOMA 21
1-5. SINGLE NUCLEOTIDE POLYMORHPISM (SNP) 21
CHAPTER II-1. INTEGRIN ALPHA V POLYMORPHISMS IN KOREAN POPULATION ARE ASSOCIATED WITH SUSCEPTIBILITY TO CHRONIC HEPATITIS B AND HEPATOCELLULAR CARCINOMA 24
ABSTRACT 25
INTRODUCTION 27
PATIENTS AND METHODS 31
PATIENTS 31
SAMPLE PREPARATION 32
SELECTION OF THE ITGAV AND RELEVANT SNPs 34
GENOTYPING OF ITGAV SNPs IN KOREAN PATIENTS 34
STATISTICAL ANALYSIS 36
RESULTS 38
DISCUSSION 49
CHAPTER II-2. GENETIC ASSOCIATION BETWEEN COLLAGEN TYPE III ALPHA I AND CHRONIC HEPAPTITIS B AND LIVER CIRRHOSIS IN KOREAN 53
ABSTRACT 54
INTRODUCTION 56
MATERIALS AND METHODS 58
PATIENTS 58
DNA PREPARATION 62
GENOTYPING METHOD FOR SINGLE NUCLEOTIDE POLYMORPHISMS 62
CONSTRUCTION OF COL3A1 PROMOTER REPOTRO VECTORS 63
RESULTS 68
GENOTYPES AND ALLELE FREQUENCIES 68
Allelic and Genotypic Association 75
Haplotype Association 78
Diplotype Association 80
Transcriptional activity by COL3A1 SNPs 84
DISCUSSION 88
CHAPTER II-3. FUNCTIONAL GENETIC VARIATIONS OF CARBONIC ANHYDRASE II IN A KOREAN POPULATION ARE ASSOCIATIED WITH INCREASED RISK OF HEPATOCELLULAR CARCINOMA 93
ABSTRACT 94
INTRODUCTION 96
PATIENTS AND METHODS 100
PATIENTS and SAMPLE PREPARATION 100
SELECTION OF THE CARBONYL ANHYDRASE AND SNPs 100
DISCOVERY OF VARIATIONS IN REGULATORY REGION OF CAII 100
GENOTYPING OF CAII SNPs 102
CONSTRUCTION OF CAII PROMTER REPORTER VECTORS 105
LUCIFERASE REPORTER GENE ASSAY 107
STATISTICAL ANAYSIS 107
RESULTS 110
GENOTYPE DISTRIBUTION 110
GENOTYPE ASSOCIATION 113
LINKAGE DISEQUILIBRIUM OF OF CAII 114
HAPLOTYPE ASSOCIATION OF CAII 119
DIPLOTYPE ASSOCIATION OF CAII 123
COX RELATIVE HAZARDS ANALYSIS FOR CAII SNPS, HAPLOTYPE, AND DIPLOTYPE 123
LUCIFERASE REPORTER ASSAYS IN PORMOTER RGION SNPS 130
DISCUSSION 134
REFERENCE 138
Table I-1. Drugs associated with chronic drug-induced liver injury (DILI) in a Spanish registry 20
Table I-2. Priorities for single nucleotide polymorphism 22
Table II-1-1. Clinical profiles of HBV clearances, chronic infection, and hepatocellular carcinoma 33
Table II-1-2. Distribution of genotypes and Hardy-Weinberg equilibrium of ITGAV polymorphisms 39
Table II-1-3. Associations between ITGAV polymorphisms with chronic infection and hepatocellular carcinoma 41
Table II-1-4. Frequencies of ITGAV haplotypes 44
Table II-1-5. Analysis of association in ITGAV haplotypes 45
Table II-1-6. Cox relative hazards analysis for age of HCC and chronic infection occurrence as functions of SNPs of the ITGAV 47
Table II-2-1. The clinical characteristics of subjects 60
Table II-2-2. The genotypes and Hardy-Weinberg equilibrium in COL3A1 70
Table II-2-3. Comparison of allele frequencies between Korean and other ethnic groups 74
Table II-2-4. Association of COL3A1 SNP and haplotype on logistic regression analysis 76
Table II-2-5. Frequencies of diplotype in COL3A1 81
Table II-2-6. Association of COL3A1 diplotype on logistic regression analysis 82
Table II-2-7. Association of diplotype in six SNP of COL3A1 on logistic regression analysis 85
Table II-3-1. Genotype frequencies of SNPs in CAII gene 111
Table II-3-2. Minor allele frequencies and Hardy-Weinberg equilibrium p-value of SNPs in CAII gene 112
Table II-3-3. Logistic analysis of SNPs in CAII gene 115
Table II-3-4. Correlation P-value and false discovery rate analysis of SNPs in CAII gene 117
Table II-3-5. Distribution of LD by r² in ethnic population and Korean 118
Table II-3-6. Haplotype frequency in Korean population 120
Table II-3-7. Haplotype frequency of CAII gene 121
Table II-3-8. Association of haplotype in CAII gene 122
Table II-3-9. Diplotype frequency in Korean population 124
Table II-3-10. Diplotype frequency of CAII gene 125
Table II-3-11. Association of diplotype in CAII gene 125
Table II-3-12. Cox relative hazards analysis for age of HCC according to SNPs of CAII 126
Table II-3-13. Cox relative hazards analysis for age of HCC according to haplotyes and diplotypes of CAII 128
Table II-3-14. Association of haplotype in CAII promoter region 131
Table II-3-15. Association of diplotype in CAII promoter region 132
Fig. II-1-1. ITGAV gene map, SNPs, and pairwise linkage disequilibrium (LD). 35
Fig. II-2-1. Plasmid construct of COL3A1 for the luciferase reporter gene. 65
Fig. II-2-2. The physical map, single-nucleotide polymorphisms, and pair-wise linkage disequilibrium of the COL3A1 gene. 69
Fig. II-2-3. Comparison of the haplotype blocks in the Korean with the International HapMap populations. 73
Fig. II-2-4. Transcriptional activity by SNPs of COL3A1. 87
Fig. II-3-1. Gene map of CAII variations and pair-wise linkage disequilibrium (LD). 101
Fig. II-3-2. Sequencing analysis and high resolution melting curve (HRM) method for CAII genotyping. 103
Fig. II-3-3. Plasmid constructs for CAII promter assay. 106
Fig. II-3-4. Kaplan-Meyer survival curves of CAII diplotypes. 129
Fig. II-3-5. Transcriptional activity by SNPs of CAII in SNU-449. 133