Title Page

Contents

CHAPTER Ⅰ. A combination of metformin/efavirenz/fluoxetine exhibits profound anticancer activity via a cancer cell-specific ROS amplification 11

ABSTRACT 12

INTRODUCTION 14

MATERIAL AND METHODS 16

RESULTS 21

DISCUSSION 35

REFERENCES 39

ABSTRACT IN KOREAN 43

CHAPTER Ⅱ. Study of anti-cancer activity by cancer cell-specific ROS amplification and resistance of cancer cells to ROS. 45

ABSTRACT 46

INTRODUCTION 48

MATERIAL AND METHODS 51

RESULTS 54

DISCUSSION 64

REFERENCES 67

ABSTRACT IN KOREAN 72

CHAPTER Ⅰ 9

Figure. 1. Metformin induces cancer cell-specific ROS increase via the AMPK/FOXO3a/MnSOD pathway. 22

Figure. 2. Combination of metformin, efaviren and fluoxetine amplifies cellular ROS and induces cell death in HCT-116 human colon cancer cells. 25

Figure. 3. Combination of metformin/efavirenz/fluoxetine (M+E+F) induces ROS amplification via synergistic inhibition of mitochondrial membrane potential and... 27

Figure. 4. Effect of drug combination M+E+F on the levels of DNA damage-induced apoptosis-, autophagy- and necroptosis-related proteins in HCT-116 and HDF cells. 29

Figure. 5. Effect of drug combination M+E+F on the levels of DNA damage-induced apoptosis-, autophagy- and necroptosis-related proteins in HCT-116 and HDF cells 30

Figure. 6. Effect of drug combination M+E+F on the levels of DNA damage-induced apoptosis-, autophagy- and necroptosis-related proteins in HCT-116 and HDF cells 31

Figure. 7. Combination of metformin/efavirenz/fluoxetine (M+E+F) effectively reduces tumor growth in an in vivo model 33

Figure. 8. A hypothetical diagram depicting mechanism of cancer cell death induced via cancer cell specific ROS amplification by combination of metformin/efavirenz/fluoxetine. 34

CHAPTER Ⅱ 10

Figure. 1 Isolation of resistant colonies of AsPc-1 and HCT116 cancer cells to MA and MEF there growth inhibition. 55

Figure. 2. The combination of (M+E+F) and (M+A) amplifies cellular ROS and decreases cell death in the AsPc-1 pancreatic cancer cell line but not in the without-resistance cancer cell line. 57

Figure. 3. In contrast to the resistant cancer cell line, the combination of (M+E+F) and (M+A) decreases cellular ROS and stops cell death in the HCT116 colon cancer cell line. 58

Figure. 4. Antioxidant activity of resistance cells 59

Figure. 5. The Protein Level of FOXO3a and Related Expression 61

Figure. 6. According to si-FOXO3a RNA transfection, FOXO3a activation plays an essential role in determining the amount of ROS in cells. 62