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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 阮雪芬(Hseuh-Fen Juan) | |
| dc.contributor.author | Yu-Wei Chang | en |
| dc.contributor.author | 張佑瑋 | zh_TW |
| dc.date.accessioned | 2021-06-13T00:07:31Z | - |
| dc.date.available | 2012-07-31 | |
| dc.date.copyright | 2007-07-31 | |
| dc.date.issued | 2007 | |
| dc.date.submitted | 2007-07-28 | |
| dc.identifier.citation | 參考文獻
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/28411 | - |
| dc.description.abstract | 在過去的幾十年問，HL一60這株細胞株是一種非常廣泛應用來研究分化治療應用在M3次分類的的急性原骨髓型白血病以及造血幹細胞分化成巨噬細胞的模式細胞。12一鄰一四葵酸一佛波醇一13一乙酸（TPA）是一種甘油二醋（DAG）的類似物，能f夠活化蛋白質激醃C進而促使HL一60細胞進行分化並失去分裂增生的能力。過去幾年有不少研究透過去氣核醣核普酸微陣列以及蛋白質體學來探討找尋什麼樣的機制牽涉到12一鄰一四葵酸一佛波醇一13一乙酸誘導HL一60細胞株分化的過程。在此，我們利用分離細胞膜與細胞質片段的蛋白質透過二維電泳及質譜儀的技術將之分離來研究蛋白質體學中非常重的次細胞區域學。我們在二維電泳膠上發現在細胞質片段有三個點其表現量是受到12一鄰一四葵酸一佛波醇一13一乙酸抑制，有四個在細胞質片段上以及八個在細胞膜片段上的點表現量受到12一鄰一四葵酸一佛波醇一13一乙酸誘導。利用穿膜區域預測軟體以及次細胞區域預測軟體進行預測，結果顯示有四種蛋白是位在粒線體、有一種蛋白質是位在內質網還有一種穿膜蛋白質則位在溶醃體。我們推測粒線體在HL一60分化的過程中扮演很重要的角色。一款micr0RNA預測軟體Milink也許能夠提供一些線索來探討這些蛋白表現變化量，透過即時定量聚合醃連鎖反應我們偵測到miR27b及miR13oa在HL一60分化過程中受到抑制。 | zh_TW |
| dc.description.abstract | The human myeloid leukemia cell line HL-60 is a useful model to study the molecular mechanisms of differentiation therapy in M3 subtype acute promyelocytic leukemia (APL) and hematopoietic development of macrophage in decades. 12-O-tetradecanoy-phorbol-13-acetate (TPA), acting via its cellular receptor protein kinase C (PKC), induce HL-60 cells to differentiate into macrophages and lose their proliferative activity. Many approaches such as DNA microarray and proteomics study have been used to find out what kinds of mechanisms are involved in the TPA-induced HL-60 differentiation process. Here we show the results of localizomics study, a very important part of proteomics study, by extracting cytoplasimc and membrane proteins from TPA-treated HL-60 cells and then following by 2DGE and mass spectrometry techniques. We found that three down-regulated and four up-regulated proteins localized in cytoplasm and eight up-regulated proteins localized in membrane. Using transmembrane domain prediction programs and subcellular localization prediction program, pWolf, 60% proteins are found in mitochondria as non-transmembrane proteins, and 40% proteins are found in lysosome and ER as non-transmembrane and transmembrane proteins, respectively. As the results showed above, we suggest that proteins in mitochondria may play an important role in the process of differentiation. Milink, a microRNA prediction program, may provide us some hints. We also show that miR27b and miR130a were down-regulated in TPA-induced HL-60 differentiation into macrophage. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-13T00:07:31Z (GMT). No. of bitstreams: 1 ntu-96-R94b43023-1.pdf: 3112581 bytes, checksum: e9a2176658b4a2a100577068b2b6c73f (MD5) Previous issue date: 2007 | en |
| dc.description.tableofcontents | 口試委員會審定書 I
誌謝 II 中文摘要 III 英文摘要 IV 第一章 緒論 1 1.1 白血病的介紹與治療現況及HL-60細胞株 1 1.2 TPA與PKC pathway 7 1.3 二維電泳(two-dimensional gel electrophoresis) 11 1.4 次細胞位置(subcellular location) 13 5 生物資訊軟體預測. . . . . . . . . ． … … 15 6 微型核醣核普酸（ microRNA ) . . . . . . ．16 7 即時定量聚合嗨連鎖反應（ real 一 time PCR ) . ． … … 17 8 動機與目的． . . . . . . . . . . ． … … 17 第二章實臉流程 . . . . . . . . ． … … 19 第三章實臉藥品與儀器. . . . . . . . . . . . . 20 3 . 1 藥品及材料. . . . . . . . . . . . . ． … … 20 3 . 1 . 1 細胞培春 . . . . . . . . . . . . . . . . . 20 3 . 1 . 2 蛋白質體學． . . . . . . .. . . . . . . . . .21 3 . 1 . 3 萃取總核糖核酸實臉 . . . . . . . ． … … 22 3 1 . 5 微型核糖核酸實臉 . . . . . . . . . . . ． … … 22 3 . 2 儀器 . . . . . . . . . . . . . . . . . . 22 3 2 . 1 細胞培春． . . . . . . 22 3 . 2 . 2 蛋白質體學 .. . . . . . . . . . ． … … 23 3 . 2 . 3 微型核糖核酸實臉． . . . . . . ． … … 23 第四章實臉方法．. . . . . . . . . . ． … … 24 4 . 1 細胞培春.. . . . .. . . . . ． … … 24 4 . 1 . 1 HL60 解凍 . . . . . . . . ． … … 24 4 . 1 . 2 細胞計數．. . . . ． … … 24 4 . 1 . 3 細胞繼代．. . . . . . . . . ． … … 24 4 . 1 . 4 細胞的實臉操作（控制對照組） ． … … 24 4 . 1 . 5 細胞的實臉操作（加藥實臉組） . . . . . . ..24 4 . 2 蛋白質體學技術．. . . . . . . ． … … 25 4 . 2 . 1 蛋白質萃取（ Protein Extraction 4）25 4 . 2 . 2 蛋白質濃度定量（ Protein Assay )25 4 . 2 . 3 二維電泳技術（黝。一 Dimensional ElectrophoresiS 26 一維等電點聚焦（ First Dimensional Isoelectric focusing , IEF ) . 26 4 . 3 微型核糖核酸即時定量聚合嗨連鎖反應 . . . . … 29 4 . 3 . 1 萃取總核糖核酸. . . . . . . . . . . . ． … … 29 4 . 3 . 2 反轉錄聚合嗨鏈鎖反應 . . . . . . . . . . . . 30 4 . 3 . 3 即時定量聚合嗨鏈鎖反應 . . . . .. . . . . . . . . . . . . . . . 31 第五章結果與討論.. . . . .. . . . . . . . . . . ． 33 5 . 1 12 一鄰一四葵酸一佛波醇一 13 一乙酸（ TPA ）誘導 HL 一 60 分化並擁有類似巨噬細胞的外表型．. . . . . . . . . . . . . . . . ． … … 33 5 . 2 次細胞層次蛋白萃取並用二維電泳分離．. . . . . . . . . . . ． … … 33 5 . 3 次細胞位置、穿膜區域與訊號肚肚軟體預測結果． . . . . . . . . . . . 40 5 . 4 次細胞層次蛋白表現變化量與互補去氣核醣核普酸微陣列相比較之結果. . . . . . . . . . . . . . . . . . 66 5 . 5 微小核醣核普酸預測軟體 milink 預測結果與即時定量聚合嗨聯鎖反應結果. .. . . . . . . . . . . . . . . . . . 74 第六章討論. . .. . . . . . . . . . . . . . . . . . . 76 6 . 1 雙硫鍵異構嗨與 HL-60 細胞秣分化後外表型改變關係的探討76 6 . 2 丙峒酸激嗨與 HL 一 60 細胞秣分化後能量代謝關係的探討 … … 76 6 . 3 角蛋白與 HL 一 60 細胞秣分化外表形、發炎反應問關係的探討 . . . . . . . . . . . . . . . . ． … … 77 6 . 4 過氣化骨髓嗨與 HL 一 60 細胞秣分化後獲得殺菌功能關係的探討. . . . . . . . . . . . ． … … 79 6 . 5 抑制素與 HL 一 60 細胞秣分化後細胞生長受到抑制的探討 … … 80 6 . 6 硫氣化還原蛋白與 HL 一 60 細胞秣分化後細胞生長受到抑制的探討 . . . . . . . . . . . . . . . . . . ． … … 81 6 . 7 熱休克蛋白 60 與 HL 一 60 細胞秣分化後細胞凋亡的探討． . ． … … 81 6 . 8 應激蛋白 70 與 HL 一 60 細胞秣分化後細胞生長受到抑制的探討 . . . . . . . . . . . . . . . . . . . ． … … 82 第七章結論．. . . . . . . . . . . . ． … … 84 參考文獻． . . . . . . . . . . . ． … … 85 | |
| dc.language.iso | zh-TW | |
| dc.title | 探討佛波酯誘導人類骨髓癌細胞HL-60分化之次細胞區域蛋白變化以及微型核醣核酸miR103a與miR27b表現的作用 | zh_TW |
| dc.title | Localizomics study in TPA-induced HL-60 differentiation into macrophages and the effects of microRNAs miR130a and miR27b. | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 95-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 黃宣誠,陳水田 | |
| dc.subject.keyword | HL60,TPA,二維電泳,微小核醣核,磷酸化蛋白, | zh_TW |
| dc.subject.keyword | HL-60,TPA,Two-dimension gel electrophoresis,microRNA,subcellular location, | en |
| dc.relation.page | 89 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2007-07-30 | |
| 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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