Title Page
Contents
Abstract 12
Introduction 15
Materials and Methods 33
Cell Culture 33
Transfection 33
Quantitative real-time PCR 33
Western blot analysis 34
Immunofluorescence 35
Wound-healing assay 35
Matrigel invasion assay 36
F-actin staining and filopodia quantification 36
Statistical analysis 37
Results 39
AGR2 exerts influence on dystroglycan protein expression but not dystroglycan gene expression in breast cancer cells. 39
Overexpression of AGR2 did not change the expression of matrix-metalloproteinases. 51
AGR2 accelerates cell migration and invasion of breast cancer cells. 57
Knockdown of dystroglycan reduces cell migration and invasion of breast cancer cells. 66
AGR2 affects cell migration and invasion through dystroglycan expression. 73
AGR2 and dystroglycan are intimately connected to cell migration and invasion through filopodia formation. 78
Discussion 86
Reference 91
국문요약 103
Table 1. List of human PDI family members 19
Table 2. Primer sequence used in this study 38
Figure 1. The architecture of AGR2 protein. 20
Figure 2. Emerging roles of AGR2 protein in cancer conditions. 21
Figure 3. Scheme of dystroglycan structure and binding protein. 27
Figure 4. Proteolytic processing of dystroglycan. 28
Figure 5. Scheme of dystroglycan retrograde trafficking. 30
Figure 6. Schematic representation of the interaction between AGR2... 32
Figure 7. AGR2 upregulation affects dystroglycan protein expression... 41
Figure 8. AGR2 protein expression level modulates the protein level... 42
Figure 9. The effect of AGR2 overexpression on dystroglycan... 44
Figure 10. The effect of AGR2 overexpression on dystroglycan... 46
Figure 11. AGR2 overexpression does not affect the dystroglycan... 48
Figure 12. AGR2 upregulation is unrelated to the gene expression... 50
Figure 13. AGR2 overexpression is unrelated to the protein and gene... 53
Figure 14. AGR2 upregulation is not involved with the protein... 56
Figure 15. Effect of AGR2 overexpression on cell migration in MDA-... 59
Figure 16. AGR2 overexpression promotes cell invasion in MDA-MB-... 60
Figure 17. AGR2 upregulation induces cell invasion in MCF7 cells. 61
Figure 18. Downregulation of AGR2 reduces cell invasion in MCF7 cells. 62
Figure 19. Effect of the phosphorylation level of ERK/AKT/Smad2 in... 63
Figure 20. AGR2 overexpression induces the phosphorylation of... 64
Figure 21. Downregulation of AGR2 reduces the phosphorylation of... 65
Figure 22. Effect of dystroglycan downregulation on cell migration in... 67
Figure 23. Reduced dystroglycan expression suppresses the cell... 69
Figure 24. Knockdown of dystroglycan reduces the invasive ability in... 70
Figure 25. Effect of the reduced dystroglycan on phosphorylation... 71
Figure 26. Effect of the dystroglycan knockdown on phosphorylation... 72
Figure 27. Effect of AGR2 overexpression via regulating... 75
Figure 28. AGR2 overexpression reduces cell invasion in MDA-MB-... 76
Figure 29. Induced AGR2 expression does not change the invasive... 77
Figure 30. Dystroglycan downregulation regulates filopodia formation... 80
Figure 31. Knockdown of dystroglycan reduces filopodia formation in... 81
Figure 32. Induced AGR2 expression promotes filopodia formation in... 82
Figure 33. AGR2 overexpression promotes filopodia formation in... 83
Figure 34. AGR2 overexpression does not affect any change of... 84
Figure 35. Upregulation of AGR2 does not affect any change of... 85