Title Page
Contents
Abstract 10
Ⅰ. INTRODUCTION 14
1. Research background of coronary artery disease 14
2. Introduction of research target PAI-1 gene 18
3. The aim of study 22
Ⅱ. MATERIALS AND METHODS 24
1. Study approval 24
2. Study population 25
3. Blood biochemical analysis 27
4. Genotyping of PAI-1 and miRNA polymorphisms 28
5. Statistical analysis 33
6. Cell culture 34
7. Plasmid amplification, transformation and purification 35
8. The prediction of miRNA binding 38
9. Over-expression of the PAI-1 gene in EA.hy926 cells 39
10. Quantitative real-time PCR 40
Ⅲ. RESULTS 41
1. Clinical characteristics of the study participants 41
2. Comparative analysis of genotype frequencies 44
3. Genotype combination analysis of PAI-1 and related miRNA polymorphisms 48
4. Haplotype and allele combination analysis of PAI-1 and related miRNA polymorphisms 58
5. Stratified analyses according to clinical factors with PAI-1 polymorphisms 68
6. Stratified analyses according to clinical factors with miRNA polymorphisms 74
7. Synergistic effect between polymorphisms of PAI-1 with clinical factors on CAD susceptibility 79
8. Clinical variables according to genotype of PAI-1 and miRNA polymorphisms using ANOVA 85
9. The difference PAI-1 expression levels according to PAI-1 3'-UTR 12068 polymorphism and micro RNA 101
Ⅳ. DISCUSSION 104
Ⅴ. CONCLUSION 110
REFERENCES 111
ABSTRACT IN KOREAN 123
Table 1. Information for genotyping of six PAI-1 polymorphisms and four miRNA polymorphisms. 32
Table 2. Baseline characteristics between CAD and controls. 43
Table 3. Genotype frequencies of PAI-1 polymorphisms in CAD and controls. 46
Table 4. Genotype frequencies of PAI-J related miRNA polymorphisms in CAD and controls. 47
Table 5. GenmvEe combination freguellcies of PAI-1 polymorphisms in CAD patients and controls. 50
Table 6. Summary of significant PAI-1 genotype combinations with greatly increased AOR in CAD patients and controls. 53
Table 7. Summary of significant genotype combinations about PAI-1 -844 and 43 polymorphisms with other PAI-1 polymorphisms in CAD patients and controls. 54
Table 8. Genotype combination frequencies of PAI-1 related miRNA polymorphisms in CAD patients and controls. 55
Table 9. Summary of significant genotype combinations of PAI-1 and related miRNA polymorphisms in CAD patients and controls. 57
Table 10. Summary of two allele haplotype analysis of the PAI-1 -675 with other polymorphisms in CAD patients and controls. 62
Table 11. Summary of two allele haplotype analysis of the PAI-1 10692 with other polymorphisms in CAD patients and controls. 63
Table 12. Summary of two allele haplotype analysis of the PAI-1 12068 with other polymorphisms in CAD patients and controls. 64
Table 13. Summary of two and three alleles haplotype analysis of the PAI-1 43 with other polymorphisms in CAD patients and controls. 65
Table 14. Summary of alleles combination analysis of the PAI-1 related miRNA polymorphisms associated with increased CAD susceptibility. 66
Table 15. Summary of alleles combination analysis of the PAI-1 related miRNA polymorphisms associated with decreased CAD susceptibility. 67
Table 16. Stratified analysis using clinical parameter with frequencies of PAI-1 genotypes. 70
Table 17. Stratified analysis using clinical parameters with frequencies of PAI-1 related miRNA genotypes. 75
Table 18. Synergistic effect of PAI-1 polymorphisms with clinical risk factor. 80
Table 19. Differences of various clinical parameters according to genotype of PAI-1 gene polymorphisms in total participants. 87
Table 20. Differences of various clinical parameters according to genotype of P4I-1 gene polymorphisms in controls. 89
Table 21. Differences of various clinical parameters according to genotype of PAI-1 gene polymorphisms 10 CAD patients. 91
Table 22. Differences of various clinical parameters according to genotype of PAI-1 related iniRNA polymorphisms in total participants. 93
Table 23. Differences of various clinical parameters according to genotype of PAI-1 related miRNA polymorphisms in controls. 95
Table 24. Differences of various clinical parameters according to genotype of PAI-1 related miRNA polymorphisms in CAD patients. 97
Figure 1. Schematic presentation of the fibrinolytic system. 21
Figure 2. Overview for polymorphisms of PAI-1 gene in this study. 23
Figure 3. Vector map inserted ORF and 3'-UTR of PAI-1 gene in pcDNA3.1(+). 37
Figure 4. LD patterns of PAI-1 gene polymorphisms. 61
Figure 5. The synergic effect analysis for interplay between clinical factor (DM, FBS, and Hgb) and PAI-1 -675 4G>5G polymorphisms in CAD susceptibility. 81
Figure 6. The synergic effect analysis for interplay between clinical factor (DM, folate, PT, and WBC) and PAI-1 10692 C>T polymorphisms... 82
Figure 7. The synergic effect analysis for interplay between clinical factor (DM, uric acid, Hgb, and neutrophils) and PAI-1 12068 G>A... 83
Figure 8. Increased AOR of PAI-1 polymorphisms with CAD risk factors. 84
Figure 9. The ANOVA analysis for difference levels of PDW according to genotype of PAI-1 43 G>A polymorphisms. 99
Figure 10. The ANOVA analysis for difference levels of neutrophils according to genotype of PAI-1 43 G>A polymorphisms. 100
Figure 11. Prediction of PAI-1 3'-UTR and miRNA binding. 102
Figure 12. PAI-1 level co-transfected with a miR-320a d and miR-330 o according to PAI-1 12068 G>A polymorphism. 103
Figure 13. Summary of significant PAI-1 and realted miRNA polymorphisms. 108
Figure 14. Schematic presentation of this study. 109