Title page
ACKNOWLEDGEMENT
Contents
ABBREVIATIONS 11
ABSTRACT 12
Ⅰ. INTRODUCTION 14
Ⅱ. MATERIALS AND METHODS 17
1. Sample 17
2. DNA Extraction 17
3. Estimation of Folate and Homocysteine Concentration 17
4. Genotyping 18
5. DNA modification 18
6. Methylation specific PCR 19
7. Statistical analysis 19
Ⅲ. RESULTS 21
1. Homocysteine metabolism-related gene polymorphisms in breast, colon and stomach cancer patient groups 21
2. Methylation status of p16INK⁴A(이미지참조), p73 and hMLH1 genes in breast, colon and stomach cancer patient groups 30
3. Association between homocysteine metabolism-related gene polymorphisms and methylation status of p16INK⁴A(이미지참조), p73, hMLH1 genes 35
4. Association between folate, homocysteine level and methylation status or polymorphisms in colon cancer patients 45
Ⅳ. DISCUSSION 52
Ⅴ. REFERENCE 56
Ⅵ. SUMMARY IN KOREAN 65
Table 1. MSP primer sequence and annealing temperature for amplication of p16INK⁴A(이미지참조), p73 and hMLH1 20
Table 2. Clinical profile in control group and breast, colon, stomach cancer patient groups 23
Table 3. Genotype distribution of MTHFR 677C〉T polymorphism in breast, colon and stomach cancer patients 24
Table 4. Genotype distribution of MTHFR 1298A〉C polymorphism in breast, colon and stomach cancer patients 25
Table 5. Genotype distribution of MTR 2756A〉G polymorphism in breast, colon and stomach cancer patients 26
Table 6. Genotype distribution of MTHFR 677C〉T polymorphism in male patients 27
Table 7. Genotype distribution of MTHFR 677C〉T polymorphism in colon cancer patients 28
Table 8. Genotype distribution of MTHFR 677C〉T and MTR 2756A〉G polymorphisms in subject of 〈65years 29
Table 9. Distribution of methylation status in control and cancer patient groups 31
Table 10. Methylation status of p16INK⁴A(이미지참조), p73 and hMLH1genes in breast, colon and stomach cancer patient groups 32
Table 11. Distribution of p16INK⁴A(이미지참조), p73 and hMLH1 methylation status in control and cancer patient groups 33
Table 12. Clinicopathological features and methylation of p16INK⁴A(이미지참조), p73 and hMLH1 genes 34
Table 13. Association between p16INK⁴A(이미지참조) methylation and homocysteine metabolism-related gene polymorphisms in breast cancer patients 36
Table 14. Association between p16INK⁴A(이미지참조) methylation and homocysteine metabolism-related gene polymorphisms in colon cancer patients 37
Table 15. Association between p16INK⁴A(이미지참조) methylation and homocysteine metabolism-related gene polymorphisms in stomach cancer patients 38
Table 16. Association between p73 methylation and homocysteine metabolism-related gene polymorphisms in breast cancer patients 39
Table 17. Association between p73 methylation and homocysteine metabolism-related gene polymorphisms in colon cancer patients 40
Table 18. Association between p73 methylation and homocysteine metabolism-related gene polymorphisms in stomach cancer patients 41
Table 19. Association between hMLH1 methylation and homocysteine metabolism-related gene polymorphisms in breast cancer patients 42
Table 20. Association between hMLH1 methylation and homocysteine metabolism-related gene polymorphisms in colon cancer patients 43
Table 21. Association between hMLH1 methylation and homocysteine metabolism-related gene polymorphisms in stomach cancer patients 44
Table 22. Plasma folate concentration associated with methylation status of p16INK⁴A(이미지참조), p73 and hMLH1genes in colon cancer patients 46
Table 23. Plasma homocysteine concentration associated with methylation status of p16INK⁴A(이미지참조), p73 and hMLH1genes in colon cancer patients 47
Table 24. Methylation status of p16INK⁴A(이미지참조), p73 and hMLH1genes in colon cancer patients according to folate level 48
Table 25. Genotypes distribution of homocysteine metabolism-related gene polymorphisms in colon cancer patients according to folate level 49
Table 26. Methylation status of p16INK⁴A(이미지참조), p73 and hMLH1genes in colon cancer patients according to homocysteine level 50
Table 27. Genotypes distribution of homocysteine metabolism-related gene polymorphisms in colon cancer patients according to homocysteine level 51
Figure 1. Homocysteine metabolism pathway and DNA methylation 16