Title Page
Contents
ABSTRACT 10
Ⅰ. INTRODUCTION 12
Ⅱ. EXPERIMENTAL SECTION 16
2.1. Meterials 16
2.2. Intrumentation 17
2.3. Detection of Hydroxyl Radical 19
2.4. Cell culture 20
2.5. Cell viability 21
2.6. Cellular Reactive Oxygen Species (ROS) Assay 22
2.7. Statistical Analysis 23
2.8. Immunocytochemistry 24
2.9. Western Blot Analysis 25
Ⅲ. RESULTS AND DISCUSSION 27
3.1. Spectroscopic Characterization of [M(N₂Py₃OH)]²⁺ Complexes. 27
3.2. Reactivity of [M(N₂Py₃OH)]²⁺ Complexes Towards Hydrogen Peroxide. 30
3.3. Cell Viability and Intracellular ROS Detection. 34
3.4. Immunocytochemistry, and Western Blot Analysis 40
Ⅳ. CONCLUSION 45
Ⅴ. REFERENCES 47
ABSTRACT IN KOREAN 51
Figure 1. Apoptosis-Ferroptosis hybrid pathway by Fenton-like reaction. 15
Figure 2. UV-vis spectra of [M(N₂Py₃OH)]²⁺ (0.25 mM, M=Mn (green), Fe (pink), Co (yellow), Ni (blue), Cu (orange), Zn (violet)) in CH₃CN at... 28
Figure 3. ESI MS spectra of [M(N₂Py₃OH)]²⁺ (M=(a) Mn, (b) Fe, (c) Co, (d) Ni, (e) Cu and (f) Zn) recorded in CH₃CN. The prominent ion peaks at... 29
Figure 4. UV-vis spectral changes of methylene blue (0.010 mM) solution during the oxidative reaction of decomposition by hydrogen... 32
Figure 5. Overlaid fluorescence spectra of [M(N₂Py₃OH)]²⁺ (M=Mn(green), Fe (pink), Co (yellow), Ni(blue) and Zn(violet))... 33
Figure 6. Effect of metal complexes on cell viability using WST-8 assay. (a) 786-O, (b) A498, (c) Caki-1 and (d) normal cell, CCD-... 37
Figure 7. (a) Effects of [Fe(N₂Py₃OH)]²⁺+ on cell viability. 786-O cells were treated with different concentrations (0, 1, 5, 10, 25, 50,... 38
Figure 8. DCFDA assay for detection of intracellular ROS produced in response to oxidative stress induced by [Fe(N₂Py₃OH)]²⁺ in 786-... 39
Figure 9. Representative images of 786-O cells NC, DW, and after incubation with [M(N₂Py₃OH)]²⁺ (M=Mn, Fe, Co, Ni, Cu, Zn) for 24... 42
Figure 10. Confocal images of nuclei and mitochondria stained by DAPI (blue) and CoX IV (green) in 786-O cells incubation with (a)... 43
Figure 11. Western blot analysis of BCL-2, p53, cleaved caspase-3 cleaved caspase-9 and GPX4 in 786 O cells after incubation with DW... 44