臺灣博碩士論文加值系統

在本項研究中，我們深入探討了天然成分Fucoidan與MTHFD2的代謝調控在不同癌症抑制中的治療策略及作用機制。我們特別探討由海藻萃取的fucoidan對大腸癌細胞之影響，以及探討在各式癌細胞中皆被過度表現的代謝酶MTHFD2於肺癌中的致癌機制進行了研究，旨在探索具治療潛力的天然藥物，以及潛力的治療標的，期盼未來在癌症治療中能發揮更顯著的治療效果。

首先我們分析了fucoidan如何透過調控基因及蛋白表現來抑制癌細胞的增生和遷移，並探討其如何促使細胞週期停滯於特定階段，以及其如何誘導癌細胞凋亡，這一系列的機制探討提供了對fucoidan抗癌作用深刻的認識。進一步，我們通過實驗結果揭示了fucoidan對於大腸癌細胞生存信號途徑的干預情況，以及其對於腸癌細胞於活體中生長能力的影響。

此外，本研究亦詳細研究了MTHFD2，一種在多種癌細胞皆過度表現的代謝酶。我們探索了MTHFD2的過度表現如何促進肺癌的生長與擴散，以及其對癌症幹細胞特性之影響，並詳細分析MTHFD2在癌症代謝重組中的角色。研究著重於MTHFD2對肺癌細胞生長和癌幹細胞特性的影響，以及它如何調節細胞內的氧化還原平衡和細胞生存機制。透過對MTHFD2於癌症代謝以及癌症幹細胞影響的深入了解，我們發現其作為一個極具潛力的癌症治療標的，期盼能對癌症治療策略提供新的方向。

綜上所述，此研究不僅深化了我們對fucoidan和MTHFD2作為癌症治療標的的認識，也為這些天然成分及代謝調控因子在未來癌症聯合治療策略中的應用提供了理論基礎及實驗證據。這些結果展現了天然成分與代謝調控在癌症治療中的重要性，為後續的研究方向和臨床試驗奠定了基礎。我們希望這些發現能夠促進更多的預臨床研究，最終為癌症患者提供更有效的治療選擇。

In this research, we investigated the interplay between the natural compound Fucoidan and the metabolic regulator MTHFD2 in inhibiting cancer development. Specifically, we analyzed Fucoidan, a compound derived from seaweed, for its potential to combat colorectal cancer. Also, we explored the pathogenic roles of MTHFD2, a metabolic enzyme notably overexpressed in various cancers, focusing particularly on lung cancer. Our goal is to unveil the anti-cancer effects of fucoidan and metabolic gene MTHFD2 to develop potential effective cancer treatment.

Our study extensively investigated fucoidan, analyzing its direct impact on colorectal cancer cells. We have systematically explored how fucoidan suppresses cancer cell growth and motility by regulating gene and protein expression, resulting in cell cycle arrest and inducing apoptosis. This comprehensive analysis elucidates the molecular mechanisms underlying fucoidan’s therapeutic effects, highlighting its potential as an anticancer agent. Furthermore, we have demonstrated fucoidan’s ability to inhibit crucial survival signaling pathways in colorectal cancer cells, thereby diminishing their proliferative and tumor formation capacities.

Furthermore, our extensive analysis on MTHFD2, which is found overexpressed in multiple cancer types, particularly emphasizes its influence in lung cancer. We investigated how MTHFD2's overexpression contributes to the proliferation and metastasis of lung cancer cells and delved into its significant part in cancer metabolic reprogramming. The research reveals the impact of MTHFD2 on energy metabolism and cellular growth signals, highlighting its regulatory capacity in maintaining cellular redox homeostasis and survival strategies. By comprehensively understanding the metabolic roles of MTHFD2, we aim to validate its suitability as a targeted approach for lung cancer therapy.

In conclusion, this result of the study not only enriches our understanding of Fucoidan and MTHFD2 as potential targets in oncology but also serves as a foundation for integrating these biological elements into possible novel cancer treatment. The findings highlight the pivotal role of natural substances and metabolic regulation in developing future cancer therapies, providing a reference for ongoing and future researches and clinical applications. We believe these studies will pave the way for further preclinical research and lead to better and more specific treatments for cancer patients. This marks a major step towards personalized and complete cancer care, drawing on insights from our research findings.

Acknowledgement i
Table of Contents ii
List of Tables vi
List of Figures vii
Abbreviations ix
中文摘要 xi
Abstract xiii
Chapter 1. Introduction 1
Global Impact of Cancer 2
Challenges in Current Cancer Therapies 2
Cancer Stemness and Metastasis 3
Metabolic Reprogramming and Redox Homeostasis 4
Potential of Traditional Chinese Medicine and Novel Compounds VMW-FC 5
Research Objectives 6
Chapter 2. Materials and Methods 8
Study 1. Anti-colorectal cancer effects of fucoidan complex-based functional beverage through retarding proliferation, cell cycle and epithelial–mesenchymal transition signaling pathways
Cell culture 9
VMW-FC Preparation 9
Cell viability 10
Colony formation assay 10
Wound-healing assay 11
Transwell migration assay 11
Cell Cycle Analysis 12
Apoptosis Detection 13
Real-time polymerase chain reaction (qPCR) 13
Western Blot 14
Animal Studies 15
Statistical analysis 16
Study 2. Modulating Redox Homeostasis and Cellular Reprogramming through Inhibited Methylenetetrahydrofolate Dehydrogenase 2 Enzymatic Activities in Lung Cancer
Cell culture 18
In silico analysis of MTHFD2 gene expression 18
Isolation of total RNA and quantitative real-time polymerase chain reaction 19
Gene knockdown by lentiviral transduction 19
Cell viability and proliferation 20
Anchorage-independent growth 21
Analysis of cell apoptosis 21
Hypoxia induction 22
Sphere formation assay 22
Western Blot 23
Cellular NADPH determination 23
Cellular ROS determination 23
In vivo limiting dilution transplantation assay 24
Statistical analysis 24
Chapter 3. Results and Discussion 25
Study 1. Anti-colorectal cancer effects of fucoidan complex-based functional beverage through retarding proliferation, cell cycle and epithelial–mesenchymal transition signaling pathways
Fucoidan complex-based functional beverage (VMW-FC) inhibits colorectal cancer (CRC) cell proliferation and cell viability 26
VMW-FC reduces CRC cell migration capability 26
VMW-FC induces the accumulation of sub-G1 phase in cell cycle 27
VMW-FC promotes apoptotic population in CRC cells 27
VMW-FC attenuates in vivo tumor growth without further adverse effects 28
VMW-FC inhibits CRC through modulating proliferation, cell cycle, apoptosis and migration-related biomarkers 29
Discussion 31
Study 2. Modulating Redox Homeostasis and Cellular Reprogramming through Inhibited Methylenetetrahydrofolate Dehydrogenase 2 Enzymatic Activities in Lung Cancer
Expression profile of MTHFD2 in lung cancer (LCa) 36
Impact of MTHFD2 knockdown on LCa growth and proliferation 36
MTHFD2-mediated lung cancer stemness 37
Impact of MTHFD2 on cellular metabolic reprogramming-dependent tumor aggressiveness 38
Cellular oxygen sensing and MTHFD2-mediated metabolic reprogramming 38
Influence of MTHFD2 on oxidative stress response and metabolic reprogramming in lung cancer 39
Discussion 40
Chapter 4. Conclusion and Perspective 45
Chapter 5. Future work 50
Chapter 6. References 54
Chapter 7. Tables and Figures 61
Chapter 8. Appendix 90
List of Tables
Table 1 62
 List of Figures
Figure 1 63
Figure 2 64
Figure 3 65
Figure 4 66
Figure 5 67
Figure 6 68
Figure 7 69
Figure 8 70
Figure 9 71
Figure 10 72
Figure 11 73
Figure 12 74
Figure 13 75
Figure 14 76
Figure 15 77
Figure 16 78
Figure 17 79
Figure 18 80
Figure 19 81
Figure 20 82
Figure 21 83
Figure 22 84
Figure 23 85
Figure 24 86
Figure 25 87
Figure 26 88
Figure 27 89

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