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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 | Pei-Wei Lee | en |
dc.contributor.author | 李珮瑋 | zh_TW |
dc.date.accessioned | 2021-06-15T11:23:13Z | - |
dc.date.available | 2017-08-30 | |
dc.date.copyright | 2016-08-30 | |
dc.date.issued | 2016 | |
dc.date.submitted | 2016-08-17 | |
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/49312 | - |
dc.description.abstract | 在癌症的治療中,最被關注的議題就是藥物的專一性以及其抑制癌細胞生長的效果。圈套寡核苷酸為一具有專一性之癌症治療藥物,其子頡子頡會與目標轉錄因子的啟動子結合,透過阻擾轉錄因子信息傳導的途徑進而造成造成癌細胞凋亡。本研究設計並開發一創新的癌細胞治療藥物:雙圈套寡核苷酸藥物,透過混合STAT3及NF-kB圈套寡核苷酸使之產生協同作用提升藥物抑制效果。本研究指出在雙圈套寡核苷酸藥物中,各別的圈套寡核苷酸依舊穩定的維持B-form結構互不相干擾。此藥物透過同時阻斷STAT3及NF-kB的作用途徑成功的以低藥物濃度(=10 nM)抑制乳癌及前列腺癌細胞生長,而在雙圈套寡核苷酸藥物的作用下,癌細胞的生存率平均為維持在40%左右,與單獨轉染的組別有相當顯著的差異; 然而當雙圈套寡核苷酸藥物在正常細胞作用時,其細胞存活率仍能維持在90%以上,證明此藥物對於癌細胞是具有相當的專一性。此外,由於STAT3以及NF-kB轉錄因子互相有著複雜的交互作用,本研究透過Survivin、IkBa、Bcl-xL及Cylin-D1的基因表現進一步指出對於STAT3而言雙圈套寡核苷酸藥物會透過獨立以及依賴NF-kB的途徑(dual pathway intervention)加強癌細胞的抑制效果。本研究的最後也指出細胞中被活化的STAT3以及NF-kB表現量不同會造成雙圈套寡核苷酸抑制效果的差異,因此我們應以此作為最適化的依據,設計針對不同癌細胞作用的雙圈套寡核苷酸藥物以期達到最好的抑制效果。簡而言之,本研究由STAT3圈套寡核苷酸為出發點,利用雙途徑干擾的概念成功開發一更具治療效果的雙圈套寡核苷酸藥物,透過細胞中被活化的STAT3及NF-kB比例進行適化,成功提出一個簡單卻有效的癌症治療方法。 | zh_TW |
dc.description.abstract | Drug specificity and anti-tumor efficiency is the most concerned issue in cancer therapy. Decoy oligodeoxynucleotide (dODN) is a cancer drug which not only targets cancer cells but also has high specificity. dODNs can combine with the promoter of transcription factors (TFs) and interrupt the signal transduce pathway that lead cancer cells undergoing apoptosis. In this study, we designed and developed a novel caner treatment, the dual decoy oligonucleotide drug by transfecting STAT3 and NF-kB dODNs to the the cancer cells simutaneously that do the synergistic effect and enhance the inhibition efficiency. The study indicates that the individual dODNs of the dual dODN drugs maintain their B-form structures stably without interfering with each other. The resarch also shows that the dual dODN drugs are able to inhibit the growth of beast and prostate cancer cells by blocking both activation pathways of STAT3 and NF-kB even in a very low concentration (=10 nM). The survival rates of cancer cells caused by dual dODN drugs are around 40% which are siginificantly lower than single dODN drugs; however, the cells survival rates keep high in normal cells that refers to their high specificity to cancer cells. Besides, through the expression of Survivin, IkBa, Bcl-xL and Cylin-D1, we imply the dual dODN drugs can enhance their anti-cancer efficiency by dual pathway intervention (NF-kB dependent and indepentdent pathway). In the last part of this research, we show that the efficiency of dual dODN drugs is relavent to the amount of STAT3 and NF-kB in cancer cells which can be the foundation of optimization. In sum, the study developed and optimized a dual dODN drug on the basis of STAT3-hpdODN to improve the inhibition efficiency of cancer cells through dual pathway intervention which has the potential to be a simple and efficient cancer therapy. | en |
dc.description.provenance | Made available in DSpace on 2021-06-15T11:23:13Z (GMT). No. of bitstreams: 1 ntu-105-R03631016-1.pdf: 4148256 bytes, checksum: c9934802eb6a6a0aa0cfd8ab0565fcb2 (MD5) Previous issue date: 2016 | en |
dc.description.tableofcontents | 口試委員審定書 i
誌謝 ii 摘要 iv Abstract v 目錄 vii 圖表目錄 x 圖目錄 x 表目錄 xi 第一章 緒論 1 1.1 前言 1 1.2 研究動機 3 1.3 研究目的 7 1.4 研究架構 9 第二章 文獻回顧 10 2.1 轉錄因子STAT3與NF-kB在癌症細胞中的表現 10 2.1.1 轉錄因子STAT3在癌症細胞中的表現 10 2.1.2 轉錄因子NF-kB在癌症細胞中的表現 13 2.1.3 轉錄因子STAT3與NF-kB 的交互作用 15 2.2 抑制STAT3及NF-kB蛋白活化之圈套寡核苷酸 18 2.2.1 抑制STAT3以及NF-kB活化之策略 18 2.2.2 圈套寡核苷酸之設計概念 20 2.2.3 STAT3及NF-kB圈套寡核苷酸 24 2.3 藥物於癌症治療的協同作用及應用 26 2.3.1 藥物的制放與共運輸系統 26 2.3.2 雙途徑抑制策略 27 第三章 材料與實驗方法 31 3.1 本研究使用之材料與儀器 31 3.1.1 實驗使用之材料 31 3.1.2 儀器 34 3.2 實驗步驟 35 3.2.1 細胞培養 35 3.2.2 細胞繼代培養 35 3.2.3 細胞計數 35 3.2.4 圈套寡核苷酸之轉染 35 3.2.5 細胞存活率分析(MTT assay) 36 3.2.6 基因表現定量分析 36 3.2.7 免疫螢光染色(Immunocytochemistry) 37 3.2.8 蛋白質萃取 38 3.2.9 蛋白質定量分析 38 3.2.10 圓二色光譜分析(CD characterization)以及Tm值量測 40 3.2.11 高分辨率熔解分析(HRM analysis) 40 第四章 結果與討論 41 4.1 STAT3圈套寡核苷酸於癌細胞之作用 41 4.1.1 STAT3圈套寡核苷酸之癌細胞抑制效率 41 4.1.2 STAT3圈套寡核苷酸之細胞選擇性 45 4.2 STAT3/NF-kB雙圈套寡核苷酸藥物之開發 47 4.2.1 雙圈套寡核苷酸藥物之結構分析 47 4.2.2 雙圈套寡核苷酸之共轉染效率 52 4.3 雙圈套寡核苷酸之作用 54 4.3.1 雙圈套寡核苷酸之癌細胞抑制效果 54 4.3.2 雙圈套寡核苷酸之細胞凋亡試驗 60 4.3.3 雙圈套寡核苷酸之作用機制 64 4.3.4 雙圈套寡核苷酸之調控 68 4.4 STAT3/NF-kB雙圈套寡核苷酸藥物之最適化 71 4.4.1 STAT3及NF-kB混合比例對於細胞存活率之影響 71 4.4.2 細胞中STAT3/NF-kB之含量與最適化 75 第五章 結論細胞脂蛋白 77 5.1 研究成果 77 5.2 未來展望 78 參考文獻 79 附錄 96 | |
dc.language.iso | zh-TW | |
dc.title | STAT3/ NF-kB雙圈套寡核苷酸藥物設計與癌細胞抑制研究 | zh_TW |
dc.title | On the design of STAT3/ NF-kB dual decoy oligonucleotide drug for cancer cell inhibition | en |
dc.type | Thesis | |
dc.date.schoolyear | 104-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 徐駿森,侯詠德,廖泰慶 | |
dc.subject.keyword | 圈套寡核?酸藥物,STAT3,NF-kB,雙途徑干擾,乳癌,前列腺癌, | zh_TW |
dc.subject.keyword | decoy oligonucleotide drug,dODN,STAT3,NF-kB,dual pathway intervention,breast cancer,prostate cancer, | en |
dc.relation.page | 98 | |
dc.identifier.doi | 10.6342/NTU201602608 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2016-08-18 | |
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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