Title Page
Contents
ABSTRACT 14
PART 1. Research Background 19
1.1. Skin cancer 20
1.1.1. Overview of Skin cancer 20
1.1.2. Melanoma 25
1.2. Apolipoprotein L1 36
1.3. Purpose of the study 41
PART 2. Systematic multiomics analysis of APOL1 expression in skin cutaneous melanoma 42
2.1. INTRODUCTION 43
2.2. MATERIALS AND METHODS 46
2.2.1. APOL1 gene expression analysis 46
2.2.2. Kaplan-Meier (KM) survival curves 46
2.2.3. Gene mutation analysis 47
2.2.4. Gene ontology analysis 48
2.2.5. Immune cell infiltration analysis 48
2.2.6. Statistical analysis 50
2.3. RESULTS 57
2.3.1. APOL1 mRNA expression levels in various types of cancer 57
2.3.2. Correlation between APOL1 mRNA expression levels and patient's survival rates in SKCM 61
2.3.3. APOL1 genome alterations 67
2.3.4. Analysis of co-expressed genes with APOL1 in SKCM 70
2.3.5. Correlation between APOL1 mRNA expression and immune cell infiltration 73
2.3.6. Correlation between APOL1 mRNA expression and effector cells in SKCM 76
2.3.7. Correlation between APOL1 mRNA expression and chemokines in SKCM 82
2.3.8. APOL1 correlated-proteins in SKCM 86
2.4. DISCUSSION 88
PART 3. Antitumor effects of APOL1 in human melanoma cells via the p38/JNK MAPK and Wnt/β-catenin signaling pathways 93
3.1. INTRODUCTION 94
3.2. MATERIALS AND METHODS 99
3.2.1. Cell culture and plasmid vector 99
3.2.2. Cell transfection 99
3.2.3. Cell proliferation assay 100
3.2.4. Colony formation assay 100
3.2.5. RT-PCR 101
3.2.6. Apoptosis assay 102
3.2.7. Cell cycle assay 102
3.2.8. Wound healing assays 103
3.2.9. Transwell cell migration and invasion assay 103
3.2.10. Western blot analysis 104
3.2.11. ELISA assays 105
3.2.12. Chemotaxis assays 105
3.2.13. In vivo tumorigenesis and metastasis assays 106
3.2.14. Immunohistochemistry 106
3.2.15. Statistical analysis 107
3.3. RESULTS 109
3.3.1. Generation of APOL1 overexpressed in A375sm human melanoma cell lines 109
3.3.2. APOL1 inhibits cell proliferation in A375sm cells 113
3.3.3. APOL1 promotes apoptosis in A375sm cells 116
3.3.4. APOL1 involves cell migration and invasion in A375sm cells 120
3.3.5. APOL1 regulates cell migration via the p38/JNK pathway in A375sm cells 123
3.3.6. APOL1 upregulates of CXCL10 in necessary for NK cell migration 128
3.3.7. APOL1 inhibits human melanoma cell A375sm tumor growth 130
3.3.8. APOL1 inhibits human melanoma cell A375sm lung metastasis 132
3.4. DISCUSSION 134
SUMMARY AND CONCLUSION 139
REFERENCE 142
ABSTRACT IN KOREAN 160
Table 1. Tumor abbreviations 51
Table 2. T-test result of HG-U133_Plus_2 using GENT2 53
Table 3. Statistical tests used and corresponding figure numbers 54
Table 4. Cox regression result for APOL1 gene expression 63
Figure 1. Main types of skin cancer 22
Figure 2. Main types of melanomas 27
Figure 3. Progression of melanoma 30
Figure 4. The major signaling pathways in melanoma 32
Figure 5. APOL1 structural domain 38
Figure 6. The biomarker of apolipoproteins in cancers 40
Figure 7. APOL1 mRNA expression level in various types of cancer and... 60
Figure 8. Correlation between APOL1 mRNA expression and overall patient survival rates in various... 64
Figure 9. Clinicopathological parameters associated with overall... 65
Figure 10. APOL1 gene alterations in SKCM 68
Figure 11. Association between APOL1 gene alterations and overall survival in SKCM 69
Figure 12. Top 50 genes positively associated with APOL1 71
Figure 13. Gene ontology analysis of the APOL1 gene in SKCM 72
Figure 14. Correlation between APOL1 mRNA expression and immune... 75
Figure 15. Correlation between APOL1 mRNA expression and immune... 78
Figure 16. Correlation between APOL1 mRNA expression and CD8+ T...[이미지참조] 79
Figure 17. Correlation of APOL1 mRNA expression with cytolytic molecule gene in SKCM 81
Figure 18. Correlation of chemokine expression with APOL1 mRNA... 85
Figure 19. Protein-Protein Interaction network of APOL1 87
Figure 20. A scheme of the xenograft model 108
Figure 21. Map of APOL1 vector and identification of stably transfected cells 110
Figure 22. APOL1 mRNA expression in transfected A375sm cells 111
Figure 23. APOL1 protein expression in transfected A375sm cells 112
Figure 24. Inhibitory effects of APOL1 on the proliferation of A375sm cells 114
Figure 25. APOL1 induces cell cycle arrest at the G1 phase in A375sm cells 115
Figure 26. APOL1 over expression induces A375sm cell apoptosis 118
Figure 27. Effects of APOL1 overexpression on the level s of apoptosis-... 119
Figure 28. APOL1 overexpression inhibits migration and invasion in... 121
Figure 29. APOL1 knock-down induces migration and invasion in... 122
Figure 30. APOL1 overexpression reduces migration in A375sm cells... 125
Figure 31. The effects of p38/JNK pathway-related protein expression... 126
Figure 32. The effects of Wnt/β-catenin pathway-related protein... 127
Figure 33. APOL1 increases the expression of CXCL10 in A375sm cells 129
Figure 34. APOL1 overexpression inhibits tumor growth 131
Figure 35. APOL1 overexpression inhibits lung metastasis 133