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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 陳林祈 | |
dc.contributor.author | Chun-Shen Chen | en |
dc.contributor.author | 陳俊昇 | zh_TW |
dc.date.accessioned | 2021-05-15T17:54:02Z | - |
dc.date.available | 2019-08-14 | |
dc.date.available | 2021-05-15T17:54:02Z | - |
dc.date.copyright | 2014-08-14 | |
dc.date.issued | 2014 | |
dc.date.submitted | 2014-07-29 | |
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/5233 | - |
dc.description.abstract | 信息傳遞及轉錄激活蛋白3(signal transducer and activator of transcription 3, STAT3)為細胞當中重要的一個轉錄因子,STAT3的活化時促使許多的細胞激素及生長因子表現。STAT3的磷酸化為其活化的原因,磷酸化後的STAT3會以二聚體之方式結合並藉由輸入蛋白(importin)送至細胞核中。進入細胞核之STAT3會與DNA結合進而造成下游基因的表現。STAT3及其所調控的基因皆在細胞的生長及存活上扮演重要的角色,而STAT3在許多腫瘤當中有持續過度表現的現象,故此調控STAT3在癌細胞中的表現及具有研究的價值。在許多的調控方法中,誘導寡核苷酸STAT3-decoy oligonucleotide(STAT3-decoy ODN)為一段能與STAT3的DNA鍵結區域結合之短序列核苷酸,與STAT3-decoy ODN結合後的STAT3無法與輸入蛋白結合進入細胞核中,藉此抑制STAT3下游基因的表現。在本論文中,我們選擇利用帶正電微脂體DOTAP/DOPE作為輸送STAT3-decoy ODN的載體,微脂體與細胞膜皆為脂質所構成,故可藉由融合或是胞吞作用將藥物送至細胞當中,而帶正電之脂質有助於將帶負電的STAT3-decoy ODN吸附於表面,增加輸送的效率。我們以乳癌細胞株MCF-7為輸送目標,探討STAT3-decoy ODN以帶正電微脂體送入細胞後在基因表現、細胞凋亡及細胞存活率的影響。我們發現,當我們利用4 μg至16 μg的STAT3-decoy ODN與32 μg微脂體所形成的脂質複合物溶於100 μl的培養液中能夠降低STAT3下游基因cyclin-D1的基因表現量並且誘導細胞的死亡,並且在六小時內即有藥性的反應且此藥性更能維持長達三天之久。 | zh_TW |
dc.description.abstract | Signal transducer and activator of transcription 3 (STAT3) is an important transcription factor, many cytokine and growth factor will be induced when STAT3 is activated. Activation of STAT3 is through enzymatic phosphorylation. Phosphorylated STAT3 trends to form a dimer structure and be sent to nucleus by importin then binds to specific DNA motif to regulate expression of downstream gene, which play a key role in cell-proliferation, metastasis and apoptosis. To regulate STAT3, STAT3-decoy oligonucleotide (STAT3-decoy ODN) is a consensus sequence with STAT3 binding motif and could trap STAT3 to restrict it transported into nucleus and silence downstream gene expression. In this thesis, cationic liposome DOTAP/DOPE is used to transport STAT3-decoy ODN into cell. Liposome is composed by lipid, which also the formation of the cell membrane. Liposome can be uptaken by cell through fusion or endocytosis. With cationic lipid, liposome can attract anionic STAT3-decoy ODN to form a lipoplex increasing gene delivery efficiency. Using MCF-7 as the target cell to study the effect of the STAT3-decoy ODN transported by cationic liposome from cell viability and gene expression. Liposome formed by 4 μg to16 μg STAT3-decoy ODN with 32 μg liposome in 100 μl medium could down-regulate gene expression of cyclin-D1, which is the downstream gene regulated by STAT3 and induce cell death during 6 hours after treatment, and the effect could last at least 3 days long. | en |
dc.description.provenance | Made available in DSpace on 2021-05-15T17:54:02Z (GMT). No. of bitstreams: 1 ntu-103-R01631020-1.pdf: 6855400 bytes, checksum: d7dbc4d2fd793142b30405667af72362 (MD5) Previous issue date: 2014 | en |
dc.description.tableofcontents | 致謝 1
中文摘要 2 Abstract 3 目錄 4 圖目錄 7 表目錄 9 第一章 緒論 10 1.1. 背景 10 1.2. 動機 14 1.3. 目的 15 1.4. 實驗架構 16 第二章 文獻回顧 17 2.1. 訊息傳遞及轉錄激活蛋白3與乳癌的關係 17 2.2. 使用decoy-ODN調控訊息傳遞及轉錄激活蛋白3的表現量 20 2.3. 以微脂體作為藥物輸送的載體 23 2.4. 不同比例的微脂體與藥物所形成之脂質複合物的輸送效率探討 25 第三章 材料與方法 28 3.1. 使用儀器與藥劑 28 3.1.1. 材料 28 3.1.2. 儀器 30 3.2. STAT3-dODN與核酸引子 32 3.3. 微脂體製備 33 3.4. 表面電位及粒徑大小 34 3.5. 細胞培養 34 3.6. 細胞繼代培養 34 3.7. 細胞計數 35 3.8. MTT assay 35 3.9. RNA萃取 35 3.10. 逆轉錄聚合酶鏈式反應 (reverse transcription-PCR, RT-PCR) 36 3.11. 即時聚合酶鏈鎖反應 (quantitative real time polymerase chain reaction , RT-qPCR)… 37 3.12. 流式細胞術 (flow cytometry) 39 3.13. 共軛焦顯微鏡 (confocal microscope) 39 第四章 實驗結果 40 4.1. 微脂體的物理性質 - 粒徑與DNA最大可攜帶量 40 4.1.1. 微脂體粒徑 40 4.1.2. 脂質複合物的DNA最大攜帶量 42 4.2. 不同比例之脂質複合物對於細胞存活率之影響 43 4.3. 時間對於脂質複合物毒殺細胞的影響 47 4.4. 以流式細胞儀鑑定脂質複合物毒殺細胞之特性 48 4.5. 由共軛焦顯微鏡觀察脂質複合物對於細胞的輸送效率 52 4.6. 以脂質複合物投藥與MCF-7後對於其基因表現的影響 54 第五章 結論與未來展望 55 5.1. 結論 55 5.2. 展望 57 第六章 參考文獻 58 | |
dc.language.iso | zh-TW | |
dc.title | 應用陽離子型微脂體基因輸送系統進行誘餌寡核苷酸誘導乳癌細胞MCF -7凋亡之研究 | zh_TW |
dc.title | A Study of Decoy Oligonucleotide-induced Breast Cancer Cell MCF-7 Apoptosis Using a Cationic Liposome Gene Delivery System | en |
dc.type | Thesis | |
dc.date.schoolyear | 102-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 劉?睿,吳嘉文,侯詠德 | |
dc.subject.keyword | 微脂體,訊息傳遞及轉錄激活蛋白3,STAT3-誘導寡核?酸,細胞死亡, | zh_TW |
dc.subject.keyword | liposome,Signal transducer and activator of transcription 3,STAT3-decoy oligonucleotide,cell death, | en |
dc.relation.page | 65 | |
dc.rights.note | 同意授權(全球公開) | |
dc.date.accepted | 2014-07-29 | |
dc.contributor.author-college | 生物資源暨農學院 | zh_TW |
dc.contributor.author-dept | 生物產業機電工程學研究所 | zh_TW |
顯示於系所單位: | 生物機電工程學系 |
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