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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 | Kuo-Chin Ni | en |
dc.contributor.author | 倪國欽 | zh_TW |
dc.date.accessioned | 2021-06-13T00:12:00Z | - |
dc.date.available | 2012-07-30 | |
dc.date.copyright | 2007-07-30 | |
dc.date.issued | 2007 | |
dc.date.submitted | 2007-07-26 | |
dc.identifier.citation | [1] Yamaguchi, H., Haranaga, S., Widen, R., Friedman, H., Yamamoto, Y., Chlamydia pneumoniae infection induces differentiation of monocytes into macrophages. Infect Immun 2002, 70, 2392-2398.
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/28554 | - |
dc.description.abstract | 癌症促進劑12-鄰-四葵酸-佛波醇-13-乙酸(TPA)是人類前骨髓血癌(APL)細胞的潛在分化誘導劑。我們整合了cDNA微陣列技術與計算生物學來研究由TPA所誘導的HL-60細胞分化。近年來的研究指出微型核糖核酸(miRNA)能夠控制細胞分化、細胞生長、細胞發育,及細胞凋亡。而微型核糖核酸是一種大約22個核苷酸長度的內生性核糖核酸,可與對應基因的mRNA作用,導致mRNA的切除或造成轉錄抑制,進而抑制基因的表現。因此,為了了解這個調節機制,很重要的是必須找出與表現量差異的基因相關的miRNA。而我們開發的軟體,miLink,即能夠建立miRNA與基因表現的關係,而預測的結果,則可透過Q-PCR進一步驗證。藉由這個研究,我們連結了在人類前骨髓性細胞分化為巨噬細胞的過程中,miRNA與基因表現的關係,並提出一個在血癌治療中,具有重要性與深遠的影響。 | zh_TW |
dc.description.abstract | The tumor promoter 12-O-tetradecanoylphorbol-13-acetate (TPA) is a potent inducer in differentiation of human acute promyelocytic leukemia (APL) cells. We applied an approach integrating cDNA microarray and computational biology to study the differentiation of HL-60 cells induced by TPA. Recent evidences have shown that MciroRNAs (miRNAs) control differentiation, cell growth, development and apoptosis. MiRNAs are endogenous ~22-nt RNAs that repress gene expression by targeting mRNA for cleavage or translational repression. Hence, for understanding the regulatory mechanism, it’s critical to identify miRNAs correlated to the differential expressed genes. Our developed software, miLink, can provide the function in building the correlation between miRNAs and gene expression. The expression results of miRNAs were confirmed by real-time Q-PCR. This study provides a connection between gene expression and microRNA of human myeloid leukemia cells differentiation to macrophages and set forth a constructive far-reaching impact on leukemia therapy. | en |
dc.description.provenance | Made available in DSpace on 2021-06-13T00:12:00Z (GMT). No. of bitstreams: 1 ntu-96-R94b43011-1.pdf: 13363766 bytes, checksum: 9383a12219c2bd8d16eefc5ae7edd6fa (MD5) Previous issue date: 2007 | en |
dc.description.tableofcontents | 口試委員審定書 i
誌謝 ii 英文摘要 iv 第一章 文獻探討及研究動機 1 1.1 血癌(Leukemia) 1 1.2 HL-60細胞株(HL-60 cell line) 4 1.3 12-鄰-四葵酸-佛波醇-13-乙酸(12-O-Tetradecanoylphorbol-13-acetate,TPA) 6 1.4 微型核糖核酸(microRNA) 8 1.5 微陣列晶片(microarray) 11 1.6 即時定量聚合酶鏈鎖反應(Real-Time PCR) 15 1.6 微陣列資料分析 22 1.6.1 GeneSpring GX 22 1.6.2 EDGE 23 第二章 實驗流程 25 第三章 實驗藥品與儀器 27 3.1 藥品及材料 27 3.1.1 細胞培養 27 3.1.2萃取總核糖核酸實驗 28 3.1.3 反轉錄cDNA並標定螢光顏色 28 3.1.4 微陣列雜交實驗 28 3.1.5微型核糖核酸實驗 28 3.2 儀器 29 3.2.1 細胞培養 29 3.2.3 微陣列雜交實驗 29 3.2.2微型核糖核酸實驗 29 第四章 實驗方法 30 4.1 細胞培養 30 4.1.1解凍HL-60細胞並培養 30 4.1.2細胞密度估計 30 4.1.3細胞繼代培養並計數 31 4.1.4 細胞倍增時間測試 32 4.1.5 HL-60控制組 32 4.1.6 HL-60實驗組 33 4.2 萃取總核糖核酸 34 4.3 將RNA轉成cDNA並標定螢光 35 4.4 微陣列雜交實驗 36 4.5 生物資訊軟體分析 39 4.5.1 GeneSpring GX微陣列資料分析 39 4.5.2 EDGE時間序列統計分析 41 4.5.3 miLink微型核糖核酸預測 42 4.6 萃取總核糖核酸(含微型核糖核酸) 42 4.7 反轉錄聚合酶鏈鎖反應 43 4.8 即時定量聚合酶鏈鎖反應 44 5.1 細胞培養 47 5.2 細胞倍增時間測試 47 5.3 微陣列實驗 48 5.3.1 RNA品質檢驗圖 48 5.3.2 RNA品質檢驗表 49 5.3.3 微陣列雜交結果圖 50 5.3.4微陣列雜交結果分析 51 5.4 GeneSpring GX分析 52 5.4.1 懸浮細胞 52 5.4.2 貼附細胞 55 5.5 總核糖核酸(含微型核糖核酸)實驗 58 5.5.1 RNA品質檢驗圖 58 5.5.2 RNA品質檢驗表 58 5.6 即時定量聚合酶鏈鎖反應 59 5.6.1 Internal Control – U6增幅曲線 59 5.6.2 miR-18a增幅曲線 60 5.6.3 miR-21增幅曲線 61 5.6.4 miR-93增幅曲線 62 5.6.5 miR-27b增幅曲線 63 5.6.6 miR-107增幅曲線 64 5.6.7 microRNA序列 65 5.7 預測之基因表現量分析 65 5.8 預測之保守基因表現量分析 68 5.9 分化功能之保守基因表現量分析 71 5.10 預測之保守基因種子區域與表現量分析 73 第六章 討論 78 第七章 結論 81 參考文獻 83 附錄 93 | |
dc.language.iso | zh-TW | |
dc.title | 探討TPA誘導HL-60細胞分化的基因表現與微型核醣核酸之關係 | zh_TW |
dc.title | Relationship between Gene Expression and microRNA in TPA-Induced HL-60 Cell Differentiation | en |
dc.type | Thesis | |
dc.date.schoolyear | 95-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 黃宣誠(Hsuan-Cheng Huang),陳水田(Shui-Tein Chen) | |
dc.subject.keyword | TPA,急性前骨髓性血癌,HL-60,微型核糖核酸, | zh_TW |
dc.subject.keyword | TPA,acute promyelocytic leukemia (APL ),HL-60,microRNA, | en |
dc.relation.page | 115 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2007-07-29 | |
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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