Title Page
ABSTRACT
국문 초록
Contents
CHAPTER 1. INTRODUCTION 13
CHAPTER 2. METHOD AND MATERIALS 15
2.1. Reagents and antibodies 15
2.2. Breast cancer cell culture 15
2.3. Cell viability assay 16
2.4. Sub-G1 analysis 17
2.5. Immunoblot analysis 17
2.6. Live cell imaging using confocal microscopy 18
2.7. Mammosphere formation assay 18
2.8. Si-RNA for UCP2 transfection 19
2.9. Aldefluor-positivity assay 19
2.10. Intracellular ROS detection 20
2.11. Mitochondrial membrane potential assay 20
2.12. Cellular ATP level detection 21
2.13. Cellular GSH level detection 21
2.14. Animals, Xenograft Experiment 22
2.15. Immunohistochemistry (IHC) and In-Situ Localization of Apoptosis (TUNEL) 23
2.16. In vivo toxicity test 23
2.17. Statistical Analysis 24
CHAPTER 3. RESULT 25
3.1. UCP2-targeted anti-CSCs drug design, characterization and targeting system 25
3.2. Genipin, Mt-Ge, and Bo-Mt-Ge induces apoptosis in HER2-positive breast cancer cells 30
3.3. Bo-Mt-Ge leads to mitochondrial function disruption 33
3.4. Effect of genipin, Mt-Ge, and Bo-Mt-Ge on HER2/HER3/Akt-axis signaling pathways in HER2-positive breast cancer 36
3.5. Discovery of UCP2 as a new target for selective CSC eradication 38
3.6. Blunted tumor growth via suppression of CSC-like activities in trastuzumab-resistant HER2-positive breast cancer xenograft model. 42
3.7. Evaluation of major organ toxicity and normal cytotoxicity of Bo-Mt-Ge 48
CHAPTER 4. DISCUSSION 50
REFERENCES 53
SUPPLEMENTARY INFORMATIONS 57
Fig 1. UCP2-targeted anti-CSCs drug design, characterization and targeting system 28
Fig 2. Influence of genipin, Mt-Ge, and Bo-Mt-Ge on cell proliferation and apoptotic signaling 31
Fig 3. Mechanism of mitochondrial action of Bo-Mt-Ge for CSCs killing 35
Fig 4. Inhibition of UCP2 Attenuates HER2/HER3/Akt axis 37
Fig 5. Inhibition of UCP2 impairs breast cancer stemness 40
Fig 6. Anti-CSCs activities in trastuzumab-resistant HER2-positive breast cancer xenograft model. 44
Fig 7. UCP2-targeting prodrugs did not induce apoptosis in normal cells and major organ toxicity. 49