Title Page
ACKNOWLEDGEMENT
CONTENTS
ABBREVIATIONS 8
ABSTRACT 9
I. INTRODUCTION 11
II. MATERIALS AND METHODS 13
II-1. Samples 13
II-2. DNA extraction 13
II-3. DNA modification 13
II-4. Methylation specific PCR 13
II-5. Genotyping 14
II-6. Statistical analysis 14
III. RESULTS 18
III-1. Methylation Status 18
III-2. The genes of Methyl-Metaboism associated with multiple methylation status 23
III-3. The genes of Methyl-Metaboism associated with multiple methylation status of tumor suppressor genes 25
IV. DISCUSSION 30
REFERENCE 38
SUMMARY IN KOREAN 41
Table 1. MSP primer sequence and annealing temperature for amplfication of p16INK4A, p14ARF and hMLH1 16
Table 2. The clinicalpathology feature in colorectal cancer patients 19
Table 3. Distribution of p16INK4A, p14ARF and hMLH1methylation status 20
Table 4. Distribution o fp16INK4A, p14ARF and hMLH1methylation status in proximal colon 21
Table 5. Distribution of p16INK4A, p14ARF and hMLH1methylation status in distal colon 22
Table 6. MTHFR C677T polymorphism associate with hyper-methyoaton status in tumor suppressor genes 24
Figure1. Methylation status of Metylation specific PCR measurement 17
Figure2. The survival duration comparison between treated both chemotherapy and surgery, and only surgery 26
Figure3-A. The survival duration is benefits to patients with hyper-methylation 27
Figure3-B. The survival curve is patients with p14ARF methylation status 28
Figure3-C. The survival curve is patients with hMLH1 methylation status 29
Figure4. According to localization of colon, survival of hypermethylation 33
Figure5. multiple hyper-methylation with MTHFR 677CC type survival curve 34
Figure6. multiple hyper-methylation with TSER 3R3R type survival curve 35
Figure7. multiple hyper-methylation with MTR A2756G type survival curve 36
Figure8. multiple hyper-methylation with MTRR A66G type survival curve 37