목차

ABSTRACT* ii
TABLE OF CONTENTS iv
LIST Of FIGURES viii
LIST OF TABLES x
Chapter 1. Overview 1
1.1. Gastric cancer 3
1.1.1. Gastric cancer 3
1.1.2. Prevalence of gastric cancer 7
1.1.3. Causes of gastric cancer 9
1.1.4. Treatment of gastric cancer 9
1.2. Apoptosis and associated proteins 10
1.2.1. Apoptosis 10
1.2.2. AMP-activated protein kinase (AMPK) 13
1.2.3. Phosphatase and tensin homolog (PTEN) 15
1.3. Anti-cancer effects of food components 17
1.3.1. Yuja peel extracts (YPEE) 17
1.3.2. Poly-γ-glutamic acid (PGA) 19
1.3.3. Makgeolli (MK) 21
1.3.4. Gamma-oryzanol (Ory) 23
1.4. Beta-sitosterol (BS) 25
1.4.1. Rice bran 25
1.4.2. Beta-sitosterol 27
1.4.3. BS bioavailability 29
1.4.4. Anti-cancer effects of BS 30
1.5. Aim of the study 32
1.6. References 33
Chapter 2. In vitro screening of rice bran for anti-cancer activities 38
2.1. Introduction 39
2.2. Materials and methods 42
2.2.1. Reagents 42
2.2.2. Cell culture 42
2.2.3. Angiogenesis assay 42
2.2.4. Cell viability assay 43
2.2.5. Statistical analysis 44
2.3. Results 45
2.3.1. The effects of rice bran components on tube formation in HUVECs 45
2.3.2. The effects of rice bran components on cell viability in AGS cells 48
2.3.3. The effects of BS on cell viability in HUVECs 50
2.4. Discussion 53
2.5. References 57
Chapter 3. Anti-cancer effects of beta-sitosterol in AGS cells and xenograft mouse model 68
3.1. Introduction 69
3.2. Materials and methods 73
3.2.1. Reagents 73
3.2.2. Cell culture 73
3.2.3. MTT assay 74
3.2.4. Annexin V assay 74
3.2.5. Caspage3/7 activity 75
3.2.6. Mito-Potential assay 76
3.2.7. RNA interference 76
3.2.8. Western blots 77
3.2.9. Two-dimensional electrophoresis (2DE-gel) and image analysis 78
3.2.10. Tumor xenograft mouse model 79
3.2.11. Cancer biomarker antibody array analysis 79
3.2.12. Statistical analysis 80
3.3. Results 81
3.3.1. BS reduces cell viability in various gastric cancer cell line 81
3.3.2. BS induces mitochondrial membrane potential in AGS cells 83
3.3.3. BS induces apoptosis in AGS cells 85
3.3.4. BS induces caspase3/7 activity in AGS cells 87
3.3.5. BS stimulates phosphorylation of AMPK and PTEN protein expressions in AGS cells 89
3.3.6. AMPK is an upstream mediator of PTEN induction following treatment with BS 91
3.3.7. AMPK is an upstream mediator of PTEN induction following treatment with BS 93
3.3.8. Comparative protein quantitation of AGS cells treated with control or BS by 2DE-gel analysis 95
3.3.9. Validation of identified proteins 98
3.3.10. Relationship between AMPK and Hsp90 100
3.3.11. AMPK is necessary to alter the levels of PTEN and Hsp90 102
3.3.12. Differentially expressed cancer biomarkers in AGS cells treated with BS 104
3.3.13. Functional annotation 106
3.3.14. Effects of BS on the growth in AGS xenograft mouse model 109
3.3.15. Toxicity of BS on xenograft mouse model 111
3.4. Discussion 113
3.5. References 121
Chapter 4. Conclusions and recommendation 128
4.1. Conclusions and recommendation 129
국문초록 133
Supplementary 135