Selective Anticancer Effects by Oxyclozanide in Triple-Negative Breast Cancer Expressing S100A9 = S100A9를 발현하는 삼중음성유방암(TNBC)에서 Oxyclozanide에 의한 선택적 항암 효과

다국어 초록 (Multilingual Abstract)

S100A9 binds to the receptor for advanced glycation end products (RAGE), forming a complex that activates various signaling pathways in cancer cells. Accordingly, inhibitors of this interaction are potential therapeutic targets. I identified and characterized oxycloazanide, a small molecule that has selective anti-cancer effects in vitro and in vivo through the inhibition of interactions between S100A9 and RAGE in triple-negative breast cancer (TNBC) cells expressing S100A9. Based on a ProteinChip array, oxyclozanide strongly inhibited the interaction between S100A9 and RAGE and toll-like receptor 4 (TLR4). In addition, oxyclozanide inhibited the proliferation of TNBC cells in an S100A9-specific manner. This inhibition was confirmed by western blotting analysis, which indicated that oxyclozanide decreased pERK expression, but increased cleaved poly (ADP-ribose) polymerase (PARP), specifically in TNBC cells expressing S100A9. In human umbilical vein endothelial cells (HUVECs), oxyclozanide inhibited processes important for angiogenesis, e.g., basic fibroblast growth factor (bFGF)-induced endothelial cell tube formation, proliferation, and migration. Furthermore, with respect to the angiogenic switch, oxyclozanide induced thrombospondin-1 (TSP-1) expression in S100A9-expressing TNBC cells. In a xenograft animal model, oxyclozanide significantly delayed tumor growth, but also suppressed the phosphorylation of ERK and induced TSP-1 in S100A9-positive tumors. The results of this study suggest that oxyclozanide is a potential drug candidate targeting S100A9-postive triple-negative breast cancer.

목차 (Table of Contents)

• Ⅰ. Introduction 1
• 1. Structure of S100A9 1
• 2. Nomenclature of S100A9 1
• 3. S100A9 localization 3
• 4. S100A9 signaling pathway 3
• 5. S100A9 expression in various cancers 6
• 6. S100A9 expression in breast cancer 7
• Ⅱ. Materials and Methods 12
• 1. Reagents 12
• 2. Cell lines 12
• 3. Molecular docking simulation 13
• 4. ProteinChip-based S100A9-RAGE/TLR4 interaction assay 13
• 4.1. ProteinChip-based S100A9 inhibitor screening 13
• 4.2. Detection and data analysis 15
• 5. Surface plasmon resonance (SPR) 15
• 6. TNBC cell proliferation assay 15
• 7. Western blotting 16
• 8. Angiogenesis assay 16
• 8.1. HUVEC tube formation assay 16
• 8.2. HUVEC proliferation assay 17
• 8.3. Wound-healing HUVEC migration assay 17
• 8.4. Basic bFGF-induced CAM angiogenesis assay. 18
• 9. Tumor xenograft study 18
• 10. Pharmacokinetic analysis 19
• Ⅲ. Results 20
• 1. Virtual screening 20
• 2. Docked pose of oxyclozanide upon S100A9 and SPR analysis 20
• 3. Inhibitory effects of oxyclozanide on S100A9-RAGE/TLR4 interactions on the ProteinChip 23
• 4. Inhibitory effects of oxyclozanide on cell growth in TNBC cells 25
• 5. Effects of oxyclozanide on signaling molecules in TNBC cells expressing S100A9 27
• 6. Inhibitory effects of oxyclozanide on bFGF-induced endothelial cell tube formation, proliferation, and migration 30
• 7. Inhibitory effects of oxyclozanide on the growing CAM 35
• 8. Inhibitory effect of oxyclozanide in an S100A9-expressing xenograft model 38
• 9. Pharmacokinetics in mice plasma 42
• Ⅳ. Discussion 45
• Ⅴ. Reference 49