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http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/66046完整後設資料紀錄
| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 郭彥彬 | |
| dc.contributor.author | Tsai-Hua Chung | en |
| dc.contributor.author | 鍾采樺 | zh_TW |
| dc.date.accessioned | 2021-06-17T00:20:05Z | - |
| dc.date.available | 2016-09-17 | |
| dc.date.copyright | 2012-09-17 | |
| dc.date.issued | 2012 | |
| dc.date.submitted | 2012-06-25 | |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/66046 | - |
| dc.description.abstract | 奈米技術的快速發展是個備受重視的研究領域，尤其在醫藥生技（奈米醫學）的應用潛力更是無限。本論文有五個部份將分別陳述，主要是結合奈米醫學、幹細胞學與藥物生理學三大領域，並且發展以利用奈米粒子或結合幹細胞的模式應用於癌症治療，另瞭解抗癌藥物對幹細胞生理的影響，試圖發展奈米生物醫學應用之新策略。
（1）氧化鐵奈米粒子促進人類幹細胞表面生長因子受體大量表現以作為癌症治療模式之應用：氧化鐵奈米粒子在幹細胞治療應用中，具有可標定幹細胞且可透過MRI偵測及追蹤。然而，氧化鐵奈米粒子對於幹細胞的影響仍不明確，並且是個不容忽視的議題。在本論文，我們將氧化鐵奈米粒子標定幹細胞後，發現會促進幹細胞表面生長因子受體EGFR大量表現。而標定後的幹細胞趨向於腫瘤的能力以及抗腫瘤生長的情形均比未標定的幹細胞優。而這些現象起因於幹細胞大量表現EGFR而抑制了腫瘤因自泌作用產生的EGF所造成的腫瘤生長、血管新生以及因腫瘤的VEGF表現所造成的腫瘤癌化與發育。我們的結果顯示，氧化鐵奈米粒子對於幹細胞的影響將可提供一個全新的幹細胞治療之應用。 （2）探討基因轉殖於人類幹細胞以結合氧化鐵奈米粒子於癌症治療應用：我們藉由氧化鐵奈米粒子促進幹細胞移動的能力，試圖合成Dextran和CM-Dextran不同材質的氧化鐵奈米粒子，利用幹細胞來攜帶具有抑制腫瘤生長的基因，加強毒殺腫瘤的能力。根據初步結果顯示，Dextran和CM-Dextran氧化鐵奈米粒子會促進人類幹細胞EGFR的大量表現，亦提昇幹細胞本身具有趨向腫瘤的能力。我們預計，幹細胞的基因轉殖技術克服後，將進一步在動物模式中進行腫瘤毒殺測試。 （3）探討人類幹細胞傳遞磁性氧化鐵奈米粒子結合抗癌藥物釋放於腫瘤治療之應用：根據上述，利用幹細胞具有趨向腫瘤且具有毒殺腫瘤能力的應用，亦可利用幹細胞將具有抑制腫瘤生長能力的基因或藥物，攜帶到腫瘤的位置，提昇毒殺腫瘤的能力。本部份，試圖合成具有可控制藥物釋放的奈米粒子，再透過幹細胞具有移動的能力，攜帶藥物到腫瘤的位置。由於，具有抗癌藥物釋放能力的奈米粒子，在合成上具有挑戰性及困難度，因此，突破在合成上的瓶頸後，接著在後續的腫瘤毒殺作用便將可順利進行。 （4）可見光誘導之二氧化鈦奈米粒子於腫瘤毒殺作用：透過浸漬法（impregnation method）的方式改變二氧化鈦吸收光的光譜，從原本UV光增加到可見光（藍光）可吸收的範圍。改質後的二氧化鈦奈米粒子容易被黑色素瘤細胞吞入，且隨著培養的時間增加，吞入的二氧化鈦奈米粒子愈多。吞入二氧化鈦奈米粒子後的黑色素瘤細胞於照光1小時後會產生活性氧，進而攻擊細胞造成細胞凋亡的現象，因此可作為黑色素瘤細胞之癌症治療應用。 （5）YC-1可減緩惡質病現象藉由脂肪裂與脂肪生成之調控：脂肪組織的減少，主要是由於脂肪裂解速度增加以及脂肪生成能力降低所造成的現象，而這也是癌症惡質病造成體重減輕的主要特徴。有太多相關致病的訊息調控影響了脂肪組織的減少，因此有必要發展有效抑制惡質病造成脂肪減少的治療藥物。根據我們的結果證明，由於YC-1具有抗腫瘤生長等多功能的生理作用，因此，YC-1影 響了3T3-L1 preadipocytes分化時的脂肪生成、也參與了TNF-α與腫瘤細胞造成的脂肪裂解調控以及腸癌細胞CT26-WT動物模式造成的惡質病的現象。YC-1會活化Akt和ERK的調控機制，促進了3T3-L1 preadipocytes 提早分化為脂肪細胞的過程，其中也活化了PPARγ、IRα、IRS3和GLUT-4等與脂肪生成相關的蛋白質；在活體外的脂肪裂解模式中，YC-1具有抑制因TNF-α或腫瘤細胞所促進而活化ERK的訊息傳遞，使得下游PLIN的蛋白質表現佭低所造成脂肪裂解的情形。在腫瘤腸癌細胞動物模式中，YC-1能抑制因腫瘤造成血液中胰島素減少，使得能延緩體重減輕的能力。綜合上述，YC-1是個新的癌症治療藥物，不僅具有抗腫瘤生長，亦具有抗腫瘤惡質病的能力。 | zh_TW |
| dc.description.abstract | With the recent progress of nanotechnology in biomedical applications, nanomedicine is a rapidly evolving area of intensive research and has received considerable attention. In this thesis, the combination of nanotechnology or nanomedicine, stem cell biology and pharmacology is used to develop novel stem cell-based tumor therapy and new strategy for nanomedicial applications. There are five topics: (1) Iron oxide nanoparticles-induced epidermal growth factor receptor expression in human stem cells for tumor therapy. (2) The enhancement of stem cell-based gene delivery by iron oxide nanoparticles for tumor therapy. (3) Stem cell-targeting delivery of controlled drug released-nanoparticles for tumor therapy. (4) Toxic effects of titanium dioxide nanoparticles in melanoma. (5) YC-1 rescues cancer cachexia by affecting lipolysis and adipogenesis.
(1) Iron oxide nanoparticles-induced epidermal growth factor receptor expression in human stem cells for tumor therapy. Superparamagnetic iron oxide (SPIO) nanoparticles show promise as labels for cellular magnetic resonance imaging (MRI) in the application of stem cell-based therapy. However, the unaddressed concerns about the impact of SPIO nanoparticles on stem cell attributes make the feasibility of SPIO labeling uncertain. Here, we show that the labeling of human mesenchymal stem cells (hMSCs) with ferucarbotran can induce epidermal growth factor receptor (EGFR) overexpression. Labeled hMSCs with their overexpressed EGFR were attracted by tumorous EGF and more effectively migrated toward tumor than unlabeled cells, resulting in more potent intrinsic antitumor activity. Moreover, the captured binding of tumorous EGF by overexpressed EGFR of labeled hMSCs blocked EGF/EGFR signaling-derived tumor growth, tumorous angiogenesis, and tumorous VEGF expression also responsible for tumor progression and development. Our results show that the impact of SPIO nanoparticles on stem cell. attributes is not necessarily harmful but can be cleverly used to be beneficial to stem cell-based therapy. (2) The enhancement of stem cell-based gene delivery by iron oxide nanoparticles for tumor therapy. With the promotion stem cell migration toward tumor by iron oxide nanoparticles, we have been trying to synthesized dextran- or Carboxymethyl-dextran (CM-dextran) -coated SPIO nanoparticles to enhance stem cell-based gene delivery. We found that as-synthesized dextran-and CM-dextran-coated SPIO nanoparticles could induce the expression of EGFR and hence stimulate the tumor tropism of stem cells as demonstrated by in vitro migration assay. A stable transfection of stem cells with TNF-a gene is proceeding. We suggest that with the establishment of transgenic stem cells SPIO-stimulated tumor tropism would be benificical to stem cell-based gene therapy for tumor. (3) Stem cell-targeting delivery of controlled drug released-nanoparticles for tumor therapy. According to the above, SPIO-induced stem cells tropism toward tumor is also used to deliver drug-loading nanoparticles stem cells used for specific tumor targeting. Therefore, we would like to integrate the targeted delivery ability of stem cells with the carrying efficiency for anticancer drugs of nanoparticles to develop a novel drug delivery system for cancer therapy. Anticancer drugs will be encapsulated into a soft-material core (polyvinyl alcohol, PVA) and then coated with a biocompatible silica shell that can protectively construct the nanoparticles and can enhance the internalization of nanoparticles into stem cells. Moreover, a controlled release mechanism (high-frequency magnetic field, HFMF) will be installed in the nanoparticle core. After the targeted-migration of stem cells with intracellular internalized nanoparticles toward tumors, HFMF could induce the release of anticancer drugs from the nanoparticle core to kill the tumor cells. (4) Visible-light inducible carbon-modified titanium dioxide nanoparticles for tumor therapy. Visible-light inducible of carbon-modified titanium dioxide were synthesized by impregnation method, and therefore these particles are called TiO2-200 nanoparticles. The photocatalytic activity that under the irradiation of visible-light was ROS produced and was measured by reduction methyl orange assay. TiO2-200 nanoparticles are easily uptaken by melanoma cells in a time-dependent manner. By irradiation with visible-light (400-500 nm) the decrease of cell viability was measured by trypan blue and induced apoptosis was measured by TUNEL assay. Intracellular ROS were generated dependent on uptake of TiO2-200 nanoparticles in treated melanoma cells. Taken together, we suggest that the carbon-modified titanium dioxide would be ideality beneficial to therapy for tumor. (5) YC-1 rescues cancer cachexia by affecting lipolysis and adipogenesis. Loss of adipose tissue, primarily due to increased lipolysis but also to an impairment of adipogenesis, is a key feature of weight loss in cancer cachexia. Because of the myriad pathogenic signaling pathways essential for atrophy of adipose tissue, effective therapeutic agents for cachectic adipose loss are lacking and urgently needed. The authors evaluated the effects of YC-1 on adipogenesis of 3T3-L1 preadipocytes, TNF-α- and tumor-cell-induced lipolysis in 3T3-L1 adipocytes, and cachectic weight loss in colon-26 adenocarcinoma-bearing mice because YC-1 has been shown to possess versatile pharmacological actions, including anticancer activity. It was found that YC-1 promotes the differentiation of 3T3-L1 preadipocytes into adipocytes through activation of Akt and extracellular signal-regulated kinase (ERK) signaling pathways as well as activation of several adipogenic mediators, such as peroxisome proliferator-activated receptor γ(PPARγ), insulin receptor α (IRα), insulin receptor substrate-3 (IRS-3) and glucose transporter-4 (GLUT-4). In the in vitro lipolysis models, YC-1 attenuates TNF-α induced lipolysis of adipocytes by antagonizing TNF-a-mediated activation of ERK and downregulation of perilipin (PLIN). It was also found that YC-1 inhibits colon-26 adenocarcinoma cell-induced lipolysis of 3T3-L1 adipocytes. Moreover, YC-1 effectively rescues cachectic weight loss in colon-26 adenocarcinoma-bearing mice by blocking lipolysis, involving insulin. Taken together the results show that YC-1 with its anticancer and anticachexia talents is highly worth developing as a novel agent for cancer therapy. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-17T00:20:05Z (GMT). No. of bitstreams: 1 ntu-101-D97422004-1.pdf: 1959780 bytes, checksum: 9ea62673c6934c5078d8700fd8ab10df (MD5) Previous issue date: 2012 | en |
| dc.description.tableofcontents | 口試委員會審定書……………………………………………………………………VI
誌謝………………………………………………………………………………...…VII 中文摘要……………………………………………………………………………..VIII 英文摘要………………………………………………………………………………..X 圖目錄 XIII 第一部份、緒論 1 第二部份、氧化鐵奈米粒子促進人類幹細胞表皮生長因子受體大量表現以作為癌症治療模式之應用 8 第一章、研究背景與動機 8 第二章、實驗方法 9 2-1實驗材料 9 2-1-1抗體 9 2-1-2實驗藥品 9 2-2 細胞培養 9 2-2-1 人類骨髓間質幹細胞 9 2-2-2 腫瘤細胞：大腸癌細胞株 10 2-3 製備標定氧化鐵奈米粒子之幹細胞 10 2-4 西方墨點法 10 2-5 螢光物質標定幹細胞 11 2-6動物受損模式建立 11 2-7病理組織切片 11 2-8普魯士藍染色 11 2-9 觀察幹細胞存在位置 11 2-10 幹細胞移動能力試驗 12 2-11 幹細胞的細胞安全性及生長能力測定 12 2-12 血管新生測定 12 2-12-1 樣品製備 12 2-12-2 老鼠組別 13 2-12-3 血紅素測定 13 2-12-4 血管內皮細胞粘著分子染色 13 第三章、結果與討論 15 3-1活體內證明幹細胞能趨向到腫瘤細胞之能力 15 3-2活體外證明氧化鐵奈米粒子促進幹細胞趨性到癌細胞的能力 15 3-3證明氧化鐵奈米粒子標定之幹細胞能抑制癌細胞造成的血管新生 15 3-4證明氧化鐵奈米粒子標定之幹細胞能抑制癌細胞之生長 16 3-5證明氧化鐵奈米粒子標定之幹細胞能抑制癌細胞VEGF表現 17 第四章、結論與未來展望 18 第三部份、探討基因轉殖於人類幹細胞以結合氧化鐵奈米粒子於癌症治療應用 19 第一章 研究背景與動機 19 第二章、實驗方法 21 2-1實驗藥品 21 2-2氧化鐵奈米粒子之製備 21 2-3 粒徑分析與表面電位之測定 21 2-4 穿透式顯微鏡觀察 21 2-5細胞培養 21 2-5-1 人類骨髓間質幹細胞 21 2-5-2 腫瘤細胞：大腸癌細胞株 22 2-6 幹細胞毒性測試 22 2-7 西方墨點法 22 2-8 幹細胞移動能力試驗 22 第三章、結果與討論 24 3-1 粒徑及表面電位 24 3-2奈米粒子對細胞的毒性影響 24 3-3 氧化鐵奈米粒子於幹細胞之功能性分析 25 第四章、結論與未來展望 26 第四部份、探討人類幹細胞傳遞磁性氧化鐵奈米粒子結合抗癌藥物釋放於腫瘤治療之應用 27 第一章 研究背景與動機 27 第二章、實驗方法 30 2-1 實驗藥品 30 2-2 製備具有藥物的磁性氧化鐵奈米粒子 30 2-3 穿透式顯微鏡觀察 30 2-4 UV-Visible光譜儀之偵測 30 2-5 腫瘤細胞：大腸癌細胞株 30 2-6 腫瘤細胞對藥物CPT的敏感性測試 30 第三章、結果與討論 31 3-1 TEM觀察裝載藥物後氧化鐵奈米粒子之構型 31 3-2 TEM觀察裝載藥物氧化鐵奈米粒子在酸性條件之構型 31 第四章、結論與未來展望 33 第五部份、可見光誘發之二氧化鈦奈米粒子於黑色素腫瘤毒殺作用 34 第一章、研究背景與動機 34 第二章、實驗方法 35 2-1實驗材料 35 2-1-1 實驗藥品 35 2-1-2實驗設備 35 2-2 奈米粒子製備與鑑定 35 2-3 表面電位之測定 35 2-4 穿透式顯微鏡觀察 35 2-5 黑色素癌細胞培養 35 2-6 活體外之處理條件 36 2-7 TEM觀察細胞之製備 36 2-7-1 TEM細胞專用slide之製備 36 2-7-2 細胞包埋 36 2-7-3 細胞切片 36 2-8 離子體質譜分析 36 2-9 細胞型態之觀察 37 2-10 細胞存活率的測量 37 2-11 細胞凋亡偵測：利用TUNEL分析法 37 2-12 細胞內活性氧的測定 37 第三章、結果與討論 39 3-1 奈米粒子製備與鑑定 39 3-2 細胞吞噬能力 39 3-3 細胞毒性評估 39 3-4 細胞凋亡評估 39 3-5 證明TiO2-200奈米粒子可於照光後產生ROS 40 第四章、結論與未來展望 41 第六部份、YC-1可減緩惡質病現象藉由脂肪裂解與脂肪生成之調控 42 第一章 研究背景與動機 42 第二章、實驗方法 44 2-1實驗藥品 44 2-2 細胞培養 44 2-3 脂肪分化實驗 44 2-4 脂肪細胞染色分析 44 2-5 脂肪裂解實驗 45 2-6 葡萄糖攝取實驗 45 2-7 偵測細胞內三酸甘油酯含量 45 2-8 細胞毒性測定 45 2-9免疫螢光實驗分析 45 2-10 西方墨點法 46 2-11 腫瘤惡質病之動物模式 46 2-12 酵素連結免疫吸附法之偵測 46 第三章、結果與討論 47 3-1 探討YC-1於3T3-L1 preadipocytes分化為脂肪細胞之影響 47 3-2探討YC-1於TNF- | |
| dc.language.iso | zh-TW | |
| dc.subject | 奈米粒子 | zh_TW |
| dc.subject | YC-1 | zh_TW |
| dc.subject | 3T3-L1 | zh_TW |
| dc.subject | 氧化鐵 | zh_TW |
| dc.subject | 幹細胞 | zh_TW |
| dc.subject | 惡質病 | zh_TW |
| dc.subject | iron oxide | en |
| dc.subject | nanoparticles | en |
| dc.subject | stem cell | en |
| dc.subject | cachexia | en |
| dc.subject | YC-1 | en |
| dc.subject | 3T3-L1 | en |
| dc.title | 奈米生物醫學新策略開發 | zh_TW |
| dc.title | New strategy development for nanomedicine | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 100-2 | |
| dc.description.degree | 博士 | |
| dc.contributor.coadvisor | 黃東明 | |
| dc.contributor.oralexamcommittee | 顧記華,林淑宜,羅履維,陳令儀,蕭仲凱 | |
| dc.subject.keyword | 氧化鐵,奈米粒子,幹細胞,惡質病,YC-1,3T3-L1, | zh_TW |
| dc.subject.keyword | iron oxide,nanoparticles,stem cell,cachexia,YC-1,3T3-L1, | en |
| dc.relation.page | 88 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2012-06-25 | |
| dc.contributor.author-college | 牙醫專業學院 | zh_TW |
| dc.contributor.author-dept | 臨床牙醫學研究所 | zh_TW |
| 顯示於系所單位： | 臨床牙醫學研究所 | |
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