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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 阮雪芬(Hsueh-Fen Juan) | |
dc.contributor.author | Chun-Li Hou | en |
dc.contributor.author | 侯君里 | zh_TW |
dc.date.accessioned | 2021-06-15T12:40:51Z | - |
dc.date.available | 2021-08-02 | |
dc.date.copyright | 2016-08-02 | |
dc.date.issued | 2016 | |
dc.date.submitted | 2016-07-27 | |
dc.identifier.citation | Adhikary S, Eilers M (2005) Transcriptional regulation and transformation by Myc proteins. Nat Rev Mol Cell Biol 6: 635-645
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/50440 | - |
dc.description.abstract | 舊藥新用為較具有投資價值的藥物開發模式,能有效的提供治療疾病藥物的選擇。神經母細胞瘤是胚胎發育過程中因交感神經分化失敗而形成的惡性腫瘤,此癌症好發於五歲以下的兒童,是孩童常見的第三種癌症,且此癌症致死率高達五成。由於此癌症中高危險群的病患,目前可選擇使用的藥物非常少,因此尋找治療惡性神經母細胞瘤用藥,是一個急需的課題。本篇研究中我們藉由分析美國國家衛生研究院所提供的資料庫(Library of Integrated Network-based Cellular Signatures, LINCS),參考現今於臨床上,治療神經母細胞瘤的用藥,比對資料庫中上千種的小分子藥物,篩選高相似性的藥物,為後續生物實驗測試。於分析結果中發現,陶扎色替(Tozasertib)和臨床用藥截剋瘤(Crizotinib)具有高度的相似性。我們更進一步使用網路藥物資料庫(Genomics of Drug Sensitivity in Cancer database, GDSC),發現陶扎色替具有更低的IC50,及表示毒殺性效果更好。接下來我們藉由細胞實驗來驗證藥物的胞殺性,還有相關細胞生理功能的影響,包含導致細胞週期停滯、細胞聚落能力降低及細胞轉移侵襲能力變弱。再來使用兩種動物實驗模型,包含異體移植腫瘤老鼠模型(xenograft mouse model)及轉基因老鼠模型(transgenic mouse model),更再進一步確定藥物於活體上的藥效。於兩種實驗動物的結果可顯示,藥效作用均可有效抑制腫瘤生長的能力,並且對於動物存活能力也有顯著的提升。最後經由相對和絕對定量等量異位標籤技術(Isobaric Tags for Relative and Absolute Quantitation, iTRAQ),並以蛋白質體學,來探討陶扎色替的藥效作用機制為何。我們於異體移植腫瘤和轉基因老鼠模型分別定量了2,912個和2,255個蛋白質,並分別挑選出在異體移植腫瘤和轉基因老鼠模型有顯著表現差異的105個和150個蛋白質。於這兩種不同的老鼠模型中,我們使用生物資訊學方法分析表現量有顯著差異的蛋白質,均發現它們多與細胞骨架的調控有關,顯示陶扎色替會影響細胞轉移和侵襲能力。根據此研究結果,指出陶扎色替對於神經母細胞瘤有良好的抑制效果,因此我們希望此研究能夠提供未來高危險群Mycn-增生的神經母細胞瘤一個新的治療策略。 | zh_TW |
dc.description.provenance | Made available in DSpace on 2021-06-15T12:40:51Z (GMT). No. of bitstreams: 1 ntu-105-R03b43011-1.pdf: 10184859 bytes, checksum: 088b8be3336ce78ed5144b31a9023cf6 (MD5) Previous issue date: 2016 | en |
dc.description.tableofcontents | 謝辭 i
中文摘要 iii Abstract v Contents vii List of Figures x List of Tables xiii Abbreviation xiv Chapter 1 Introduction 1 1.1 Neuroblastoma 1 1.2 MYCN 2 1.3 Drug discovery 4 1.4 An Aurora kinase inhibitor: tozasertib 5 1.5 Proteomics 6 1.6 Motivation 7 Chapter 2 Materials and Methods 9 2.1 Experimental design 9 2.2 Perturbation similarity analysis 10 2.3 Cell culture 10 2.4 Drug treatment 11 2.5 Cell viability assay using MTS assay 11 2.6 Colony formation assay 11 2.7 DNA content analysis 12 2.8 Flow cytometric analysis of apoptosis using Annexin V-FITC/PI staining 13 2.9 Animal efficacy studies 13 2.10 Tumor protein preparation 15 2.11 Reduction, alkylation, and digestion of proteins 15 2.12 iTRAQ labeling of peptides 16 2.13 Strong cation exchange (SCX) chromatography 17 2.14 StageTip desalting 17 2.15 ZipTip desalting 18 2.16 LC-MS/MS analysis 18 2.17 Protein identification and quantification 20 2.18 Selection of differentially expressed proteins 21 2.19 Functional enrichment analysis and association network 22 2.20 Western blotting 22 2.21 Transwell migration and invasion assay 23 2.22 Statistical analysis 24 Chapter 3 Results 25 3.1 Drug repurposing via perturbation similarity analysis 25 3.2 Tozasertib causes cytotoxicity of neuroblastoma cell lines 25 3.3 Tozasertib reduces proliferation of neuroblastoma cell lines 26 3.4 Tozasertib induces G2/M phase arrest in neuroblastoma cell lines 26 3.5 Tozasertib induces apoptosis in neuroblastoma cell lines 27 3.6 Tozasertib has anti-cancer effect on neuroblastoma xenograft mice 28 3.7 Tozasertib has anti-cancer effect in neuroblastoma transgenic mice 29 3.8 Proteomic analysis identifies changes with tozasertib treatment in small-scale experiments 30 3.9 Proteomic analysis identifies changes with tozasertib treatment in large-scale experiments 32 3.10 Functional analysis of proteomic data 33 3.11 Tozasertib reduces metastasis of neuroblastoma cell lines 34 3.12 Pathway enrichment analysis of proteomic data 35 Chapter 4 Discussion 37 Chapter 5 Future work 46 Chapter 6 References 48 Figures 59 Tables 106 Appendix 131 | |
dc.language.iso | en | |
dc.title | 探討泛極光激酶抑制劑tozasertib於Mycn擴增之神經母細胞瘤的舊藥新用潛力 | zh_TW |
dc.title | The pan-Aurora kinase inhibitor tozasertib as a repurposed drug for Mycn-amplified neuroblastoma | en |
dc.type | Thesis | |
dc.date.schoolyear | 104-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 黃宣誠(Hsuan-Cheng Huang),黃敏銓(Min-Chuan Huang),許文明(Wen-Ming Hsu) | |
dc.subject.keyword | 惡性,Mycn基因擴增,神經母細胞瘤,陶扎色替,舊藥新用,蛋白質體學,相對和絕對定量等量異位標籤技術, | zh_TW |
dc.subject.keyword | high-risk,Mycn-amplified,neuroblastoma,tozasertib,drug repurposing,proteomics,iTRAQ, | en |
dc.relation.page | 135 | |
dc.identifier.doi | 10.6342/NTU201601400 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2016-07-28 | |
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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