세포자멸사 및 혈관신생 경로 조절을 통한 개 유선종양에 대한 나린제닌의 항암 작용 = Naringenin as a potent anti-cancer agent for canine mammary gland tumor through modulation of apoptotic and angiogenic pathways

다국어 초록 (Multilingual Abstract)

Canine mammary gland tumors (CMTs) are the most prevalent tumors in intact female dogs, and surgery is the primary option. Chemotherapy is used in inoperable patients or as adjuvant therapy after surgery. However, the clinical application of human chemotherapeutic agents to CMTs is limited by the lack of standard protocols, uncertain clinical efficacy, and substantial risk of adverse effects. Thus, there is urgent need to discover safe and effective anti-cancer agents for CMTs. Naringenin, a flavonoid derived from citrus fruits, exhibits diverse pharmacological effects. The effects of naringenin on various cancers, including breast cancer, have been studied; however, its effects on CMTs have not been investigated. Therefore, the aim of this study is to evaluate the anti-cancer effect of naringenin to CMTs. Naringenin inhibited the viability and migration of CMT-U27 canine mammary gland tumor cells and increased expression of cleaved caspase-3. In a CMT xenograft mouse model, naringenin suppressed tumor growth and elevated cleaved caspase-3 expression in tumor tissues. In addition, a reduction in tumor vessel density was observed. The anti-metastatic effect of naringenin was verified by decreased cytokeratin expression in the inguinal lymph nodes. Naringenin showed anti-angiogenic effects by inhibiting canine aortic endothelial cell migration and tube formation, while also suppressing microvessel sprouting in a rat aortic ring model. In conclusion, this study suggests that naringenin is a promising chemotherapeutic candidate for CMTs by inducing apoptosis and inhibiting angiogenesis.

목차 (Table of Contents)

• INTRODUCTION 1
• MATERIALS AND METHODS 4
• 1. Cell culture and reagent 4
• 2. 3(4,5dimethylthiazol2yl)5(3carboxymethoxyphenyl)2(4sulfophenyl)2H tetrazolium (MTS) assay 4
• 3. Annexin V/propidium iodide (PI) Staining 5
• 4. Western blotting 5
• 5. Immunocytochemistry 6
• 6. Xenograft tumor model and Naringenin treatment 7
• 7. Immunohistochemistry 7
• 8. Migration assay 8
• 9. Tube formation assay 9
• 10. Rat aortic ring assay 9
• 11. Statistical analysis 10
• RESULTS 11
• 1. Naringenin induces apoptosis in CMT-U27 cells by Caspase-3 activation 11
• 2. Naringenin inhibits tumor growth and induces Caspase-3 dependent apoptosis in vivo 12
• 3. Naringenin exerts anti-angiogenic activity 13
• 4. Naringenin inhibits CMT-U27 cell migration and lymph node metastasis 14
• DISCUSSION 26
• REFERENCES 30
• ABSTRACT (IN KOREAN) 35