Multi-omics profiling for the anti-proliferative activity of ortho-topolin riboside and protocatechualdehyde combination using in vitro and in vivo radio-resistant MDA-MB-231 cell models

다국어 초록 (Multilingual Abstract)

Triple-negative breast cancer (TNBC) poses a significant therapeutic challenge due to its intrinsic resistance to traditional hormonal and targeted therapies. Radio-resistant TNBC is one of the most serious problems in improving the outcomes of treatment. To our knowledge, the anticancer activity induced by co-treatment of ortho-topolin riboside (oTR) and protocatechualdehyde (PA) in radio-resistant TNBC cells has not been investigated. The present study aimed to investigate the potential anticancer effect of oTR and PA in in vitro and in vivo radio-resistant MDA-MB-231 (MDA-MB-231/RR) cell models. Transcriptomic, proteomic and metabolomic analyses were employed to identify major altered molecular pathways and tumor growth inhibition. The co-treatment of oTR and PA synergistically reduced the cell viability of MDA-MB-231/RR cells compared to single oTR or PA treatment. Identified major altered pathways were ‘β- alanine metabolism’, ‘hypoxia-inducible factor 1 (HIF-1) signaling’, ‘FoxO signaling’, and the ‘mitogen-activated protein kinase (MAPK) signaling’. In xenograft mouse models, a significant reduction in tumor volume was observed in both single treatment (oTR or PA alone) and co-treatment (low dose and high dose) groups with no significant alterations in body weight and liver morphology. The findings of this study demonstrate the potential of oTR and PA co-treatment as a promising therapeutic strategy for the treatment of radio-resistant TNBC.

국문 초록 (Abstract)

삼중 음성 유방암(TNBC)은 기존의 호르몬 및 표적 치료법에 대한 본질적인 내성으로 인해 심각한 치료 과제를 안고 있습니다. 방사선 내성 TNBC는 치료 결과를 개선하는 데 있어 가장 심각한 문제입니다. 우리가 아는 한, 방사선 내성 TNBC 세포에서 오르토토폴린 리보사이드(oTR)와 프로토카테츄알데히드(PA)의 병용 치료로 유도되는 항암 활성은 조사된 바 없습니다. 본 연구는 시험관 및 생체 내 방사선 내성 MDA-MB-231(MDA-MB-231/RR) 세포 모델에서 oTR과 PA의 잠재적 항암 효과를 조사하는 것을 목표로 했습니다. 전사체, 단백질체 및 대사체 분석을 통해 주요 변화된 분자 경로와 종양 성장 억제를 확인했습니다. oTR과 PA의 병용 처리는 단일 oTR 또는 PA 처리에 비해 MDA-MB-231/RR 세포의 세포 생존력을 시너지 효과를 발휘하여 감소시켰습니다. 변경된 주요 경로는 'β-알라닌 대사', '저산소증 유도 인자 1(HIF-1) 신호', 'FoxO 신호', '미토겐 활성화 단백질 키나제(MAPK) 신호'로 확인되었습니다. 이종이식 마우스 모델에서 단일 치료(oTR 또는 PA 단독) 및 병용 치료(저용량 및 고용량) 그룹 모두에서 종양 부피가 유의하게 감소했으며 체중과 간 형태에는 큰 변화가 없었습니다. 이 연구 결과는 방사선 내성 TNBC 치료를 위한 유망한 치료 전략으로서 oTR과 PA 병용 치료의 잠재력을 보여줍니다.

목차 (Table of Contents)

• I. Introduction 1
• II. Materials and methods 4
• 1. Chemicals and reagents 4
• 2. Cell culture 5
• 3. Colony formation assay 5
• 4. Cell death assay 6
• 5. Western blot analysis 6
• 6. Cell viability analysis 6
• 7. Xenograft mouse model 7
• 8. Histopathological analysis 8
• 9. Cell lysis and protein quantification 9
• 10. Metabolomic profiling analysis 10
• 11. Transcriptomic profiling analysis 11
• 12. Proteomic profiling analysis 12
• 13. Glycolytic proton efflux rate and oxygen consumption rate measurements 13
• 14. Data processing and statistical analysis 14
• III. Results 16
• 1. Characteristics of radiation-resistant breast cancer MDA-MB-231/RR cells 16
• 2. Effect of co-treatment of oTR and PA on MDA-MB-231/RR cell viability 17
• 3. Inhibition of tumor growth by co-treatment of oTR and PA in xenograft model 19
• 4. Alteration of metabolic profiles in MDA-MB-231/RR cells by co-treatment of oTR and PA 21
• 5. Alteration of transcriptomic profiles in MDA-MB-231/RR cells by co-treatment of oTR and PA 29
• 6. Alteration of proteomic profiles in MDA-MB-231/RR cells by co-treatment of oTR and PA 30
• 7. Glycolytic rate and mitochondrial respiration inhibition in MDA-MB-231/RR cells on co-treatment of oTR and PA 31
• 8. Multi-omics profiling 36
• IV. Discussion 41
• V. Conclusion 47
• References 48
• Appendices 58
• 국문초록 73