請用此 Handle URI 來引用此文件:
http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/8840
完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 張富雄(Fu-Hsiung Chang) | |
dc.contributor.author | Wei-Ju Chen | en |
dc.contributor.author | 陳薇如 | zh_TW |
dc.date.accessioned | 2021-05-20T20:02:22Z | - |
dc.date.available | 2010-09-15 | |
dc.date.available | 2021-05-20T20:02:22Z | - |
dc.date.copyright | 2009-09-15 | |
dc.date.issued | 2009 | |
dc.date.submitted | 2009-08-19 | |
dc.identifier.citation | Alexander G. Tkachenko, Huan Xie, Yanli Liu, Donna Coleman, Joseph Ryan, Wilhelm R. Glomm, Mathew K. Shipton, Stefan Franzen, and Daniel L. Feldheim (2004) Cellular Trajectories of Peptide-Modified Gold Particle Complexes: Comparison of Nuclear Localization Signals and Peptide Transduction Domains. Bioconjug. Chem., 15, 482-490.
Berry, C.C., de la Fuente, J.M., Mullin, M., Chu, S.W.L. and Curtis, A.S.G (2007) Nuclear localization of HIV-1 tat functionalized gold nanoparticles. IEEE Trans. Nanobioscience, 6, 262-9. Cartiera, M.S., Johnson, K.M., Rajendran V., Caplan, M.J. and Saltzman, W.M. (2009) The uptake and intracellular fate of PLGA nanoparticles in epithelial cells. Biomaterials, 30, 2790-2798. De la Fuente, J.M. and Berry, C.C. (2005) Tat peptide as an efficient molecule to translocate gold nanoparticles into the cell nucleus. Bioconjug. Chem., 16, 1176-80. Maysinger D. (2007) Nanoparticles and cells: good companions and doomed partnerships. Org. Biomol. Chem., 5, 2335–2342. Edwige Gros, Sebastien Deshayes, May C. Morris, Gudrun Aldrian-Herrada, Julien Depollier, Frederic Heitz and Gilles Divita. (2006) A non-covalent peptide-based strategy for protein and peptide nucleic acid transduction. Biochimica. et Biophysica. Acta., 1758, 384–393. Funnell, W.R. and Maysinger D. (2006) Three-dimensional reconstruction of cell nuclei, internalized quantum dots and sites of lipid peroxidation. J. Nanobiotec., 20, 4-10. Garden, O.A., Reynolds, P.R., Yates J., Larkman, D.J., Marelli-Berg, F.M., Haskard, D.O., Edwards, A.D. and George, A.J. (2006) A rapid method for labelling CD4+ T cells with ultrasmall paramagnetic iron oxide nanoparticles for magnetic resonance imaging that preserves proliferative, regulatory and migratory behaviour in vitro. J. Immunol. Methods, 31, 123-33. Ho, Y.P., Chen, H.H., Leong, K.W. and Wang, T.H. (2006) Evaluating the intracellular stability and unpacking of DNA nanocomplexes by quantum dots-FRET. J. Control. Rel., 116, 83-89. Jae-Hyun Lee, Kyuri Lee, Seung Ho Moon, Yuhan Lee, Tae Gwan Park and Jinwoo Cheon. (2009) All-in-One Target-Cell-Specific Magnetic Nanoparticles for Simultaneous Molecular Imaging and siRNA Delivery. Angew. Chem. Int. Ed., 48, 4174 –4179. James, J. Lu, Robert Langer, and Jianzhu Chen. (2009) A Novel Mechanism Is Involved in Cationic Lipid-Mediated Functional siRNA Delivery. Mol. Pharm., 6, 763-771. Jui-Chih Chang, Hong-Lin Su and Shan-hui Hsu. (2008) The use of peptide-delivery to protect human adipose-derived adult stem cells from damage caused by the internalization of quantum dots. Biomaterials, 29, 925–936. Liu L., Guo K., Lu J., Venkatraman, S.S., Luo D., Ng, K.C., Ling, E.A., Moochhala S. and Yang, Y.Y. (2008) Biologically active core/shell nanoparticles self-assembled from cholesterol-terminated PEG-TAT for drug delivery across the blood-brain barrier. Biomaterials, 29, 1509-17. Liu X., Wang Y., Nakamura K., Kubo A. and Hnatowich, D.J. (2008) Cell studies of a three-component antisense MORF/tat/Herceptin nanoparticle designed for tumor delivery. Cancer Gene Ther., 15, 26-32. Li, S.D. and Huang L. (2008) Pharmacokinetics and biodistribution of nanoparticles. Mol. Pharm., 5, 496-504. Maysinger D., Lovrić J., Eisenberg A. and Savić R. (2007) Fate of micelles and quantum dots in cells. Eur. J. Pharm. Biopharm., 65, 270-281. Mikhaylova M., Stasinopoulos I., Kato Y., Artemov D. and Bhujwalla, Z.M. (2009) Imaging of cationic multifunctional liposome-mediated delivery of COX-2 siRNA. Cancer Gene Ther., 16, 217–226. Nam, H.Y., Kwon, S.M., Chung H., Lee, S.Y., Kwon, S.H., Jeon H., Kim Y., Park, J.H., Kim J., Her S., Oh, Y.K., Kwon, I..C., Kim K. and Jeong, S.Y. (2009) Cellular uptake mechanism and intracellular fate of hydrophobically modified glycol chitosan nanoparticles. J. Controlled Release, 135, 259-67. Ruan G., Agrawal A., Marcus, A.I. and Nie S. (2007) Imaging and tracking of tat peptide-conjugated quantum dots in living cells: new insights into nanoparticle uptake, intracellular transport, and vesicle shedding. J. Am. Chem. Soc., 28, 14759-14766. Ryan, J.A., Overton, K.W., Speight, M.E., Oldenburg, C.N., Loo L., Robarge W., Franzen S. and Feldheim, D.L. (2007) Cellular uptake of gold nanoparticles passivated with BSA-SV40 large T antigen conjugates. Anal. Chem., 79, 9150-9159. Shu-Chen Hsieha, Fung-Fang Wangb, Ching-Shih Lina, Yu-Ju Chena, Shih-Chieh Hungc and Yng-Jiin Wanga. (2006) The inhibition of osteogenesis with human bone marrow mesenchymal stem cells by CdSe/ZnS quantum dot labels. Biomaterials, 27, 1656–1664. Tkachenko, A.G., Xie H., Liu Y., Coleman D., Ryan J., Glomm, W.R., Shipton, M.K., Franzen S. and Feldheim, D.L. (2004) Cellular trajectories of peptide-modified gold particle complexes: comparison of nuclear localization signals and peptide transduction domains. Bioconjug. Chem., 15, 482-90. Vives E., Richard, J.P., Rispal C. and Lebleu B. (2003) TAT peptide internalization: seeking the mechanism of entry. Curr. Protein Pept. Sci., 4, 125–132. Wadia, J.S., Stan, R.V. and Dowdy, S.F. (2004) Transducible TAT-HA fusogenic peptide enhances escape of TAT fusion proteins after lipid raft macropinocytosis. Nat. Med., 10, 310–315. Yanli Liu and Stefan Franzen. (2008) Factors Determining the Efficacy of Nuclear Delivery of Antisense Oligonucleotides by Gold Nanoparticles. Bioconjugate Chem., 19, 1009–1016. Ziegler A., Nervi P., Dürrenberger M. and Seelig J. (2005) The cationic cell-penetrating peptide CPP(TAT) derived from the HIV-1 protein TAT is rapidly transported into living fibroblasts: optical, biophysical, and metabolic evidence. Biochemistry, 44, 138-48. 劉逸祥. (2008) 結合 Tat 和 NLS 胜肽於磁性奈米粒子表面對細胞分佈之研究. 國立台灣大學醫學院生物化學暨分子生物學研究所碩士論文. | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/8840 | - |
dc.description.abstract | 基因治療提供了治療疾病的前景,一般來說,利用奈米粒子來攜帶藥物或治療性的分子,例如: DNA、RNA、胜肽、蛋白質等進入標的細胞,對於核酸分子而言,有兩大標的位置-細胞質與細胞核;因此如何有效地將分子穿過細胞膜以到達每個標的位置是重要的課題。由於細胞膜是個動態的結構,傾向於親脂性的分子通過,而限制了親水性或帶電荷性的大分子物質的穿透,所以核酸類的物質比較難以自由進入到細胞內,為此,發展了以非病毒型的微脂體遞送系統,配合在奈米粒子的表面修飾物,提高進入細胞的效率;本篇研究利用微脂體-氧化鐵奈米粒子表面修飾生物素與鏈黴親和素融合SV40-Tag核位訊號 (NLS) 建構之蛋白質,階段式輔以磁力將奈米磁性粒子與螢光標定之寡核苷酸送入細胞質內後,並透過NLS與細胞內分子將奈米粒子送入細胞核內。
本論文提出利用磁力代替穿透膜胜肽的方法,觀察奈米粒子在癌細胞與老鼠骨髓間質幹細胞之細胞內分佈。實驗結果為1. 施予磁力後可有效增加細胞對奈米粒子的吸收;2. 自然處理細胞後其NLS-ST蛋白質本身並不能進入細胞質,唯有加入磁力後可顯著加強細胞質中之奈米粒子進入細胞核;3. 奈米粒子進入細胞後可能脫離溶體路徑,有助於標靶奈米粒子進入細胞核。 | zh_TW |
dc.description.abstract | Gene therapy offers the promise of treating disease. In general, two target sites inside cells are cytoplasm or nucleus in case of delivering therapeutic molecules such as DNA, RNA, peptides and proteins by nanoparticles. There are complicated problems in targeting these molecules. It was difficult for the large charge molecules such as DNA to pass across plasma and nucleic membrane on their own. For these reasons, I developed a non-viral micelle delivery system using the surface-modified lipid nanoparticles. It was composed one positive charge lipid, called GEC-cholesterol with cholesterol, PEG lipid and biotinylated lipid. The surface-exposed biotin moiety could interact with the bifunctioned fusion protein containing streptavidin. Adding magnetic field sequentially could force the lipid nanoparticles complexed with oligonucleotides into the cytosol; meanwhile NLS associated with intracellular binding proteins then transported into the nucleus of cancer cells and rat bone marrow stem cells. The results indicated that manipulating the magnetic force could efficiently enhance nanoparrticles transported into cells. Besides, NLS-Streptavidin fusion proteins themselves couldn’t enter into the cytoplasm. But magnetic nanoparticles inside the lipid micelles were entered significantly into the nucleus with the assistant of magnetic fields. Thus, magnetic field might be benefit for targeting nanoparticles with cargos into nucleus. | en |
dc.description.provenance | Made available in DSpace on 2021-05-20T20:02:22Z (GMT). No. of bitstreams: 1 ntu-98-R96442012-1.pdf: 798271 bytes, checksum: e595fcf3a00936caca423572e5c9c998 (MD5) Previous issue date: 2009 | en |
dc.description.tableofcontents | 口試委員會審定書 i
謝 誌 ii 中文摘要 iii Abstract iv 圖表目錄 vii 第一章 緒 論 1 1.1 物質進出細胞的機制 1 1.2 細胞穿透性胜肽 1 1.3 細胞核運輸機制與核位訊號 2 1.4 細胞核的標靶遞送 3 1.5 奈米粒子的細胞內命運 4 1.6 研究動機 5 第二章 實驗材料和方法 6 2.1 實驗材料 6 2.1.1 化學藥品 6 2.1.2 儀器 7 2.1.3 細胞株 8 2.1.4 質體與寡核苷酸 8 2.1.5 脂質 8 2.1.6 奈米粒子 9 2.2 實驗方法 9 2.2.1 氧化鐵奈米粒子之製備 9 2.2.2 TAT-PAST與NLS-ST 的蛋白質抽取 11 2.2.3 蛋白質的定量與定性 12 2.2.4 包覆奈米磁性粒子與蛋白質混和的製備 13 2.2.5 寡核苷酸吸附於奈米粒子之盤式試驗比例分析 15 2.2.6 細胞毒性試驗 (MTT assay) 15 2.2.7 雷射共軛焦顯微鏡與數據分析 16 第三章 實驗結果 17 3.1 脂質奈米粒子之特性分析: 粒徑大小與表面電位 17 3.2 細胞生長與奈米粒子劑量毒性測試 17 3.3 奈米粒子於各個時間點在細胞內分布情形 18 3.4 磁力可以加強連接 NLS-ST 蛋白質之奈米磁性粒子遞送至細胞核 18 第四章 討 論 20 第五章 圖表說明 22 第六章 參考文獻 26 | |
dc.language.iso | zh-TW | |
dc.title | 正價微脂體攜帶寡核苷酸複合物與穿膜促進劑在哺乳類細胞內之吸收與分佈分析 | zh_TW |
dc.title | Analysis of Cellular Uptake and Distribution of Cationic Micelles Complexed with Oligonucleotides and Transmenbrane Enhancers in Mammalian Cells | en |
dc.type | Thesis | |
dc.date.schoolyear | 97-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 莊榮輝,許金玉 | |
dc.subject.keyword | 磁性奈米粒子,寡核苷,酸,細胞核遞送, | zh_TW |
dc.subject.keyword | magnetic nanoparticles,oligonucleotides,nuclear targeting, | en |
dc.relation.page | 29 | |
dc.rights.note | 同意授權(全球公開) | |
dc.date.accepted | 2009-08-19 | |
dc.contributor.author-college | 醫學院 | zh_TW |
dc.contributor.author-dept | 生物化學暨分子生物學研究所 | zh_TW |
顯示於系所單位: | 生物化學暨分子生物學科研究所 |
文件中的檔案:
檔案 | 大小 | 格式 | |
---|---|---|---|
ntu-98-1.pdf | 779.56 kB | Adobe PDF | 檢視/開啟 |
系統中的文件,除了特別指名其著作權條款之外,均受到著作權保護,並且保留所有的權利。