Abstract
CONTENTS
Ⅰ. INTRODUCTION 7
Ⅱ. MATERIALS AND METHODS 9
1. Cells and Materials 9
2. Cell count and Flow Cytometry Analysis 10
3. DNA fragmentation Assay 10
4. Caspase 3 activity Assay 11
5. RNA isolation and Reverse transcriptase-polymerase chain reaction (RT-PCR) 11
6. DNA transfection and luciferase assay 12
Ⅲ. RESULTS 13
1. Cytotoxic effect of triptolide on U937 cell death 13
2. Caspase mediates triptolide-induced apoptosis 18
3. Induction of apoptosis by triptolide appears to be independent of formation of reactive metabolites 23
4. Overexpression of Bcl-2 prevents triptolide-mediated caspase 3 activation 25
5. Modulation of Bcl-2 and IAP protein families in triptolideinduced apoptosis 28
Ⅳ. DISCUSSION 32
REFERENCES 35
KOREAN ABSTRACT 39
Figure 1. Effect of triptolide on U937 cell growth rate, viability, morphological characteristics and DNA fragmentation 14
Figure 2. Flow cytometric analysis of apoptotic cells 17
Figure 3. Effect of triptolide on caspase-specific cleavage of PLC-1, and caspase 3 activity 20
Figure 4. Effect of N-acetylcysteine (NAC) on triptolide-induced caspase 3 activity and cleavage of PLC-1 24
Figure 5. Effect of Bcl-2 overexpression on triptolide-induced apoptosis 26
Figure 6. The expression levels of apoptosis related proteins in U937 cells by treatment with triptolide 30