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Anticancer effect and cell death mechanism of phytochemicals in human gastric cancer cell lines

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Author(s)
문정용
Issued Date
2013
Awarded Date
2013. 2
Type
Dissertation
URI
http://dcoll.jejunu.ac.kr/jsp/common/DcLoOrgPer.jsp?sItemId=000000006154
Alternative Author(s)
Moon,Jeong Yong
Affiliation
제주대학교 대학원
Department
대학원 생명공학과
Advisor
김소미
Table Of Contents
Contents 1
List of Tables 6
List of Figures 7
List of Abbreviations 9

PART I. Introduction 10
1. Traditional Citrus sp. (dangyuja, phalsak, and yeagam) in Jeju 11
2. Phytochemicals 12
3. Gastric cancer 13
4. Apoptosis. 13
5. Autophagy. 14
6. Scope of the study 14

PART II. Essential Oil of jeju traditional Citrus 15
II-1. Antiproliferative activity of Essential Oils
II-2. Antiproliferative activities of major compounds of Essential Oils
1. ABSTRACT 16
2. INTRODUCTION 17
3. MATERIALS AND METHODS 18
3.1. Reagents 18
3.2. Preparation of the Citrus Essential Oils .18
3.3. Cell culture 18
3.4. Cell viability assay. 18
3.5. GC-MS analysis. 19

4. RESULTS 20
4.1. Cytotoxic effect of EOs 20
4.2. Chemical composition of Jeju Citrus essential oils. 24
4.3. Effect of major compounds of Citrus peel essential oils in human gastric cancer cells 28
5. DISCUSSION 30

PART III. Super Critical Extracts of jeju traditional Citrus 32
III-1. Antiproliferative activities of Super Critical Extracts (SCE) in AGS cells
III-2. Compositional analysis of Super Critical Extract
III-2. Antiproliferative activity of Auraptene and its Mechanism in SNU-1 Human Gastric Cancer Cell
1. ABSTRACT 33
2. INTRODUCTION 34
3. MATERIALS AND METHODS 36
3.1. Plant materials 36
3.2. Supercritical fluid extraction 36
3.3. GC-MS analysis. 36
3.4. Cell culture 37
3.5. Cell viability assay 37
3.6. Microscopic observation of cellular morphology 38
3.7. Flowcytometric analysis 38
3.8. Westernblot analysis 38
4. RESULTS 40
4.1. Cytotoxic effect of SCEs 40
4.2. Chemical constituents of Jeju Citrus Supercritical Extracts 42
4.3. Cytotoxic effect of auraptene 61
4.4. Effect of Auraptene on cell morphology, cell cycle distribution and ΔΨm 63
4.5. Effects of auraptene on the regulation of apoptosis-related proteins 65
4.6. Auraptene induces negative feedback activation of Akt signaling by inhibition of mTOR 67
5. DISCUSSION 69

PART IV Anticancer activity of Nobiletin and its Mechanism in p53-Mutated SNU-16 Human Gastric Cancer cells 71
IV-1. Nobiletin Potentiates the Effects of the Anticancer Drug 5-Fluorouracil
IV-2. Nobiletin Induces Endoplasmic Reticulum Stress-Mediated Autophagy/Apoptosis
1. ABSTRACT 72
2. INTRODUCTION 74
3. MATERIALS AND METHODS 77
3.1. Chemicals and reagents 77
3.2. Cell culture 77
3.3. Cell viability assay 77
3.4. Analysis of the effects of drug combinations 78
3.5. Cellular morphology and DNA fragmentation 78
3.6. Flow cytometric analysis 78
3.7. Western blot analysis 79
3.8. Caspase-9, -3 activity assay 79
3.9. 2-D Materials 80
3.10. Protein sample preparation 80
3.11. 2D PAGE 81
3.12. Image analysis 81
3.13. Peptide Mass Fingerprinting (PMF) 82
3.14. Statistical analysis 82
4. RESULTS 83
4.1. Nobiletin inhibits the proliferation of SNU-16 cells 83
4.2. Effect of nobiletin on cell morphology, DNA laddering, and cell cycle distribution 84
4.3. Effects of nobiletin on the expression of apoptosis-related proteins and p21 90
4.4. Synergistic effects of nobiletin and 5-FU on SNU-16 cell growth 93
4.5. p53-mutated SNU-16 cells respond to nobiletin and 5-FU via different pathways. 95
4.6. Nobiletin induce level of GRP78 protein in SNU-16 cells. 99
4.7. Nobiletin induce endoplasmic reticulum stress-mediated autophagy and apoptosis 102
4.8. Nobiletin induced autophagy through the Akt/mTRO signaling pathway in SNU-16 cells 104
4.9. Inhibition of autophagy enhances antiproliferative effects of nobiletin in SNU-16 cells 106
5. DISCUSSION 109
References 110
List of Publications 124
Degree
Doctor
Publisher
제주대학교
Citation
문정용. (2013). Anticancer effect and cell death mechanism of phytochemicals in human gastric cancer cell lines
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General Graduate School > Biomaterials Science and Technology
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