Title Page
ABSTRACT
Contents
Nomenclature 10
1. Introduction 11
1.1. Ginsenoside 11
1.1.1. Classification of Ginsenoside 11
1.1.2. Ginsenoside Rh2 12
1.1.2. Ginsenoside CK 12
1.2. Cancer 13
1.2.1. Prostate Cancer (PCa) 15
1.2.2. Colorectal Carcinoma (CRC) 15
1.2.3. Liver Cancer 16
1.2.4. Breast Cancer 16
1.2.5. Neuroblastoma 16
1.3. Apoptotic Signaling Pathway 17
1.4. Cell Cycle Arrest 19
1.5. Cancer Invasion and Metastasis 20
2. Materials and Methods 22
2.1. Rh2 and CK 22
2.2. Cell culture and GS Treatments 22
2.3. Cell Viability and IC₅₀ Assay 23
2.4. Cell Migration Assay 24
2.5. Florescence Analysis of Apoptosis/Necrosis 24
2.6. qPCR 24
2.7. Statistical Analysis 25
3. Results and Discussion 26
3.1. Rh2 26
3.1.1. Cell Viability 26
3.1.2. Cell Migration Assay 30
3.1.3. Apoptosis and Necrosis Analysis 32
3.2. CK 37
3.2.1. Cell Viability 37
3.2.2. Cell Migration Assay 40
3.2.3. Apoptosis and Necrosis Analysis 42
3.2.4. Cancer-related Gene Expression 48
4. Conclusion 53
Bibliography 55
Summary in Korean 65
Table 1.1. Common metastatic sites of certain solid tumors 20
Table 2.1. Cancer cell lines and cell culture medium used in this study 22
Table 2.2. Cell culture container and cell seeding density for each experiment purpose 23
Table 2.3. DNA sequence of gene-specific primers for qPCR used in this study 25
Table 3.1. IC₅₀ and cell migration inhibition concentration of Rh2 according to cancer cell type 30
Table 3.2. IC₅₀ concentration of CK according to each cancer cell and results of other researchers 37
Fig. 1.1. Chemical structures of the two forms of GS-Rh2. (A) Chemical structure of 20(S)-GS-Rh2; (B) Chemical structure of 20(R)-GS-Rh2. 12
Fig. 1.2. Structures and main metabolic pathways of 20(S)-Rb1, 20(S)-Rg1 and 20(S)-Rg3. GS are known to be metabolized by human intestinal microflora. This scheme represents the structures and... 13
Fig. 1.3. Ten Leading Cancer Types for the Estimated New Cancer Cases and Deaths by Sex, United States, 2022. Estimates are rounded to the nearest 10 and exclude basal cell and squamous cell skin... 14
Fig. 1.4. Apoptosis signaling pathways. Apoptosis pathways can be initiated through different entry sites, for example, at the plasma membrane by death receptor ligation (receptor pathway) or at the... 17
Fig. 1.5. Pathways of apoptosis. 18
Fig. 1.6. Schematic of the cell cycle progression 19
Fig. 3.1. Cell viability of Du145, HCT116 and Huh-7 exposed to various concentrations of Rh2 and IC₅₀ concentration of Rh2 for each cancer cell. 27
Fig. 3.2. Cell viability of MCF-7, MDA-MB-231 and SK-N-MC exposed to various concentrations of Rh2 and IC₅₀ concentration of Rh2 for each cancer cell. 29
Fig. 3.3. Cell migration assay results of Du145 and HCT116 cells treated with various concentrations of Rh2 31
Fig. 3.4. Cell migration assay results of breast cancer cells treated with various concentrations of Rh2 31
Fig. 3.5. Cell migration assay results of Huh-7 and SK-N-MC cells treated with various concentrations of Rh2 32
Fig. 3.6. Apoptosis and necrosis images of Rh2-treated Du145 cells through fluorescence staining analysis 33
Fig. 3.7. Apoptosis and necrosis images of Rh2-treated HCT116 cells through fluorescence staining analysis 34
Fig. 3.8. Apoptosis and necrosis images of Rh2-treated MCF-7 cells through fluorescence staining 35
Fig. 3.9. Apoptosis and necrosis images of Rh2-treated MDA-MB-231 cells through fluorescence staining 36
Fig. 3.10. Cell viability of Du145, HCT116 and Huh-7 exposed to various concentrations of CK and IC₅₀ concentration of CK for each cancer cell. 38
Fig. 3.11. Cell viability of MCF-7, MDA-MB-231 and SK-N-MC exposed to various concentrations of CK and IC₅₀ concentration of CK for each cancer cell. 39
Fig. 3.12. Cell migration assay results of Huh-7 and SK-N-MC cells treated with various concentrations of CK 40
Fig. 3.13. Cell migration assay results of Du145 and HCT116 cells treated with various concentrations of CK 41
Fig. 3.14. Cell migration assay results of breast cancer cells treated with various concentrations of CK 41
Fig. 3.15. Apoptosis and necrosis images of CK-treated Du145 cells through fluorescence staining 43
Fig. 3.16. Apoptosis and necrosis images of CK-treated HCT116 cells through fluorescence staining 44
Fig. 3.17. Apoptosis and necrosis images of CK-treated Huh-7 cells through fluorescence staining 45
Fig. 3.18. Apoptosis and necrosis images of CK-treated MCF-7 cells through fluorescence staining 46
Fig. 3.19. Apoptosis and necrosis images of CK-treated MDA-MB-231 cells through fluorescence staining 47
Fig. 3.20. Expression of genes related to the apoptosis signaling pathway in each cancer cell treated with CK 49
Fig. 3.21. Expression of genes related to the cell cycle in each cancer cell treated with CK 50
Fig. 3.22. Expression of genes involved in cell proliferation in each cancer cell treated with CK 51
Fig. 3.23. Expression of genes involved in tumor invasion and metastasis in each cancer cell treated with CK 52