Title Page
Abstract
Contents
List of abbreviations 8
1. Introduction 9
2. Materials and methods 10
2.1. Antibodies (Abs) and drugs 10
2.2. Cell culture 11
2.3. Cell viability assay 11
2.4. Cell proliferation assay 12
2.5. Immunofluorescence staining 12
2.6. Caspase-3 activity 13
2.7. Annexin V FITC and propidium iodide (PI) apoptosis detection assay 13
2.8. Cell cycle arrest 13
2.9. Cell-derived mouse tumor xenografts model 14
2.10. Immunohistochemical staining 15
2.11. Quantitative real-time PCR analysis 15
2.12. Protein preparation and western blot 16
2.13. Immunoblot analysis 16
2.14. Statistical analysis 17
3. Results 17
3.1. BRD4 overexpression in ovarian cancer cells and half-maximal inhibitory concentration of OPT-0139 17
3.2. OPT-0139's inhibitory effect on cell survival and proliferation 19
3.3. OPT-0139's induction of apoptotic cell death 20
3.4. OPT-0139's induction of cell cycle arrest 21
3.5. OPT-0139's inhibition of tumorigenesis in a mouse xenograft model 22
3.6. OPT-0139 alters hypoxia signaling, angiogenesis, cancer stemness, and apoptosis gene expression levels in tumors 24
3.7. Combining OPT-0139 with cisplatin in ovarian cancer cells 26
3.8. Combining OPT-0139 with cisplatin in a mouse xenograft model 27
4. Discussion 31
Bibliography 35
초록 (국문) 38
Figure 1. BRD4 expression in various ovarian cancer cell lines, SKOV3, A2780, OVCAR3, CAOV3, and HOSEpic. (A) Western blots and (B) relative... 18
Figure 2. Inhibition of cancer cell survival and proliferation by OPT-0139: SKOV3 and OVCAR3 were treated with OPT-0139 for 48 h at each... 20
Figure 3. Induction of apoptotic cell death by OPT-0139: The SKOV3 and OVCAR3 cells were treated with the indicated doses of OPT-0139 for 48 h,... 21
Figure 4. Cell cycle arrest induction by OPT-0139: OPT-0139 effects cell cycle phase distribution of (A) SKOV3 and (B) OVCAR3 cells (*** p 〈0.001... 22
Figure 5. Anticancer effect of OPT-0139 in mouse xenograft models: (A) Tumor growth rate with OPT 0139 low-dose and high-dose injection in... 24
Figure 6. Relative mRNA level measurement: (A) BRD4, Bromodomain-containing protein; (B) HIF-1a, Hypoxia-inducible factor; (C) Bcl-2, b-cell... 25
Figure 7. Additive effect of combining OPT-0139 with cisplatin in ovarian cancer cell: (A) Cell viability, (B) Caspase-3 activity, (C) Western blot.... 27
Figure 8. Additive effect of combining OPT-0139 with cisplatin in mouse xenograft models: mice were injected with cisplatin 2 mg/kg (intravenous)... 29
Figure 9. Relative mRNA levels between the control, cisplatin only, OPT-0139 only, and OPT-0139 and cisplatin combination groups: (A) BRD4,... 30