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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 阮雪芬(Hsueh-Fen Juan) | |
| dc.contributor.author | Jia-Wei Hsu | en |
| dc.contributor.author | 許家維 | zh_TW |
| dc.date.accessioned | 2021-06-13T15:18:44Z | - |
| dc.date.available | 2013-07-30 | |
| dc.date.copyright | 2008-07-30 | |
| dc.date.issued | 2008 | |
| dc.date.submitted | 2008-07-23 | |
| dc.identifier.citation | Almog, N. and V. Rotter (1997). Involvement of p53 in cell differentiation and development. Biochim Biophys Acta 1333, F1-27.
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/37072 | - |
| dc.description.abstract | 靈芝在許多亞洲國家中已被視為可以促進身體健康及延年益壽的中草藥，而經由靈芝所純化、萃取出的多醣體在許多過去的研究中証實，其具有調控免疫反應及抗腫瘤的功效。翁啟惠院士研究團隊將靈芝萃取物多醣體中的主要活性成分純化出，並將之命名為F3。在我們過去的研究中，F3可以促使人類單核球白血病細胞株THP-1分泌許多細胞激素，例如TNF-a。這促使我們思考：靈芝對於抗腫瘤療效的作用機制及分子調控為何？因此，我們整合了微陣列基因晶片分析以及基因網路建構，試著去了解F3如何在THP-1細胞株中產生抗腫瘤的效用。在微陣列基因晶片資料分析中，我們找出在生物統計上具有顯著意義的生物路徑及蛋白質交互作用網路，其中「死亡接受器誘導細胞凋亡」的生物路徑是非常具有顯著差異，而在細胞凋亡相關細胞實驗也可以發現凋亡細胞的產生，同時我們進一步利用即時定量聚合酶連鎖反應及西方墨點法驗證此一生物路徑。此外，THP-1單核球白血病細胞株在F3的刺激之下還會分化為巨噬細胞，從細胞的貼附、細胞週期停止、氮藍四銼還原、以及巨噬細胞分化標記的大量表現可以證實這個現象。另外，我們也分別利用細胞凋亡蛋白酵素caspase及轉錄因子p53的抑制劑，證實了caspase的切割以及p53的活化參與在單核球分化為巨噬細胞的過程當中。透過系統生物學分析方法，我們的研究結果顯示F3對於人類單核球白血病細胞株THP-1造成細胞凋亡及分化為巨噬細胞的生物效應，以及其相關作用之分子機制，期許將來可以應用於白血病的治療上。 | zh_TW |
| dc.description.abstract | Ganoderma lucidum has long been used to modulate the immune system in preventing or treating various human diseases in Asian countries. The biologically active compounds originally isolated and purified from Ganoderma lucidum were identified as polysaccharides, and the main fraction was designated as F3. In our study, F3 can activate many cytokines such as TNF-a in human leukemia cell line THP-1. Here, we integrated time-course microarray analysis and network modeling to study the F3-induced effects on THP-1 cells. In the microarray data analysis, we identified the differentially disturbed pathways with statistical significance based on Fisher’s exact test and false discovery rate. The pathway of apoptosis induction through death receptors is shown to be very significant in F3-treated THP-1 cells. Apoptosis-related assays were performed to show the apoptotic changes in F3-treated THP-1 cells. Caspase-3, 7, 8, 9 were cleaved into their active forms after F3 treatment. In addition, dynamic gene expression profiles showed that treatment of THP-1 cells with F3 might be capable of inducing cell differentiation. F3-treated THP-1 cells have capacity of enhancing macrophage differentiation, demonstrated by changes of cell adherence, cell cycle arrest, nitroblue tetrazolium reduction, and expression of differentiation markers including CD11b, CD14, CD68, matrix metalloproteinase-9, and myeloperoxidase. Furthermore, caspase cleavage and p53 activation were found to be significant in F3-induced THP-1 cells. General caspase inhibitor and p53 inhibitor reduced the percentage of F3-treated THP-1 cells to express CD11b and CD14. Altogether, these data indicated that F3 is able to induce cell death via death receptor signaling and differentiation of THP-1 cells into macrophages by activating caspase cascade and p53. Our results provide a molecular explanation for the properties of F3 on human leukemia cells by systems biology approach and put forth a valuable prospect for leukemia and cancer therapy. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-13T15:18:44Z (GMT). No. of bitstreams: 1 ntu-97-R95b43001-1.pdf: 9028147 bytes, checksum: 1b8bff46788e682dc00bbc0f0fc6cc36 (MD5) Previous issue date: 2008 | en |
| dc.description.tableofcontents | Abstract
1. Introduction 1 1.1 Leukemia 1 1.2 Treatment Strategies for Leukemia 2 1.2.1 Cell Death 2 1.2.2 Cell Differentiation 4 1.3 Ganoderma lucidum (Reishi or Lingzhi) 6 2. Specific Aim 8 3. Materials and Methods 9 3.1 Lab instruments and equipments 9 3.2 Antibodies and chemicals 11 3.3 Experimental procedures 14 4. Results 21 4.1 Microarray analysis of overall gene expression in F3-induced THP-1 cells 21 4.2 Apoptosis induction of F3 on THP-1 cells 22 4.3 Validation of death receptor signaling in F3-induced apoptosis 23 4.4 Significantly functional networks and biological pathways that contribute to F3-induced THP-1 cells 23 4.5 Potentiation of F3-induced THP-1 cells differentiation into macrophages 24 4.6 Expression of specific macrophage differentiation markers 25 4.7 Monocytic THP-1-derived macrophages association proteins after F3 induction 26 4.8 Regulation of F3-induced macrophage differentiation 27 5. Discussion 29 6. Conclusion 33 7. References 34 8. Figures 44 9. Appendices 64 Appendix I. Purification and main structure of G. lucidum polysaccharides F3. 64 Appendix II. Significant Gene Set Analysis by Ingenuity Pathway Analysis (IPA) web tool. 65 Appendix III. Significantly genetic networks affected by F3 69 Appendix IV. The protein-protein interaction network as a framework for cell death. 71 Appendix V. The protein-protein interaction network as a framework for cell differentiation. 73 Appendix VI. Selected significant pathways by Ingenuity Pathway Analysis (IPA) web tool. 75 Appendix VII. Selected significant pathways by ArrayXPath. 81 Appendix VIII. Total RNA quality check by agarose gel. 82 Appendix IX. Quantification and melting curve of gene expression by Q-PCR. 83 Appendix X. Raw data of Ct value in gene expression patterns by Q-PCR. 85 Appendix XI. Product check of the Q-PCR products using agarose gel. 86 Publication 87 | |
| dc.language.iso | en | |
| dc.subject | 分化 | zh_TW |
| dc.subject | 靈芝 | zh_TW |
| dc.subject | 多醣體 | zh_TW |
| dc.subject | 單核球白血病細胞 | zh_TW |
| dc.subject | 細胞凋亡 | zh_TW |
| dc.subject | apoptosis | en |
| dc.subject | Ganoderma lucidum | en |
| dc.subject | differentiation | en |
| dc.subject | polysaccharides | en |
| dc.subject | monocytic leukemia cells | en |
| dc.title | 靈芝多醣體誘導細胞凋亡及分化之系統生物學研究 | zh_TW |
| dc.title | Systems Analysis Reveals Apoptosis and Macrophage Differentiation Induced by Ganoderma lucidum Polysaccharides | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 96-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 黃宣誠(Hsuan-Cheng Huang),陳水田(Shui-Tein Chen) | |
| dc.subject.keyword | 靈芝,多醣體,單核球白血病細胞,細胞凋亡,分化, | zh_TW |
| dc.subject.keyword | Ganoderma lucidum,polysaccharides,monocytic leukemia cells,apoptosis,differentiation, | en |
| dc.relation.page | 42 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2008-07-25 | |
| dc.contributor.author-college | 生命科學院 | zh_TW |
| dc.contributor.author-dept | 分子與細胞生物學研究所 | zh_TW |
| 顯示於系所單位： | 分子與細胞生物學研究所 | |
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