Programmable RNA Nanoflowers Engineered via Rolling Circle Transcription for Synergistic and Targeted Cancer Therapy = 롤링 서클 전사를 이용한 다기능 RNA 나노플라워 구축을 통한 시너지 기반 암 치료

목차 (Table of Contents)

• Ⅰ. Introduction 1
• 1. Overview 1
• 2. RNAi 3
• a. RNAi mechanism 3
• b. The targets of RNAi in tumor treatment 5
• 3. Targeted Delivery System for Tumor RNAi Therapy 10
• a. Liposomes 11
• b. Polymer-based delivery system 13
• c. Inorganic nanoparticles 16
• d. Organic metal frameworks 18
• e. Biomolecules 20
• 4. Multifunctional NAHs 22
• a. Construction of NAHs 22
• b. Applications of NAHs in biomedicine 25
• 5. Application of NAHs Incorporating RNAi therapeutics 31
• a. RNAi-mediated gene therapy combined with chemotherapy 32
• b. RNAi-mediated gene therapy combined with PDT 33
• c. RNAi-mediated gene therapy combined with PTT 35
• d. RNAi-mediated gene therapy combined with immunotherapy 37
• 6. Research ideas and contents of this thesis 39
• Ⅱ. Dual-Targeting RNA Nanoflowers Constructed through Rolling Circle Transcription as a Synergistic Nanoplatform for Breast Cancer Therapy 42
• 1. Introduction 42
• 2. Experimental section 44
• a. Reagents 44
• b. Instrument 44
• c. Preparation of circular DNA templates 45
• d. RCT and preparation of RNA nanoflowers 46
• e. Aptamer functionalization and self-assembly of RNA nanoflowers 46
• f. Cell resuscitation, culture, and cryopreservation 46
• g. Cellular uptake assays 47
• h. Cytotoxicity assay (CCK-8 assay) 48
• i. Apoptosis detection 48
• j. Real-time quantitative PCR (qPCR) analysis 49
• k. Establishment of tumor-bearing mouse model 50
• l. In vivo imaging experiments 50
• m. Tissue staining analysis 51
• n. Statistical analysis 51
• 3. Results and discussion 51
• a. Principle of the experiment 51
• b. Synthesis and optimization of NF 52
• c. Stability of NF 57
• d. Synthesis of NF-Chol@DOX 59
• e. In vitro uptake and targeting assessment of NF-Chol 68
• f. Evaluation of the synergistic therapeutic efficacy of NF-Chol@DOX 71
• g. Silencing effect of NF-Chol@DOX on BCRP and MUC1 76
• h. In vivo targeting and tissue distribution of NF-Chol 77
• i. In vivo therapeutic efficacy of NF-Chol@DOX 79
• j. Biocompatibility of NF-Chol@DOX 80
• 4. Conclusion 85
• Ⅲ. RNA Nanoflowers Derived from Rolling Circle Transcription for Photothermal/Immunotherapeutic Synergistic Cancer Treatment 86
• 1. Introduction 86
• 2. Experimental section 88
• a. Reagents 88
• b. Instrument 88
• c. Preparation of circular DNA templates 89
• d. RCT and preparation of RNA nanoflowers 89
• e. Preparation of RNF@PEI/ICG-Apt 90
• f. Cell resuscitation, culture, and cryopreservation 90
• g. Cellular uptake assays 91
• h. Cytotoxicity assay (CCK-8 assay) 91
• i. Apoptosis detection 92
• 3. Results and discussion 92
• a. Principle of the experiment 92
• b. Synthesis and optimization of RNF 94
• c. Synthesis and optimization of RNF@PEI 95
• d. Synthesis of RNF@PEI/ICG-Apt 96
• e. Photothermal properties of RNF@PEI/ICG-Apt 101
• f. In vitro uptake and targeting assessment of RNF@PEI/ICG-Apt 103
• g. Cytotoxicity of RNF@PEI/ICG-Apt 105
• 4. Conclusion 106
• Ⅳ. Summary and Prospect 107
• 1. Summary of work 107
• 2. Prospect of work 108
• References 111
• Acknowledgments 159
• Korean Abstract 161