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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 陳志毅(Jyh-Yih Chen) | |
dc.contributor.author | Yi-Chun Chen | en |
dc.contributor.author | 陳儀君 | zh_TW |
dc.date.accessioned | 2021-06-08T00:49:23Z | - |
dc.date.copyright | 2015-07-21 | |
dc.date.issued | 2015 | |
dc.date.submitted | 2015-07-13 | |
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/18042 | - |
dc.description.abstract | 傳統外科手術、荷爾蒙及標靶藥物均難以有效治療三陰性 (ER/PR/HER2) 的轉移性惡性乳癌。三陰性乳癌細胞雖然對於傳統化療藥物有良好的反應,但是化療藥物對於正常細胞與癌細胞之間並無區別性,反而會造成癌細胞產生抗藥性並且具有極大副作用。因此尋找化療藥物的分子標的將有助於開發對於三陰性乳癌細胞具有專一性毒性之藥物。陽離子抗菌胜肽對於乳癌細胞與正常細胞之間具有選擇性的毒殺作用。Tilapia piscidin 4 (TP4) 為Nile tilapia(Oreochromis niloticus)所分泌的一種陽離子抗菌胜肽。人工合成之TP4能有效抑制革蘭氏陽性及陰性菌的生長,但對於是否具有毒殺癌細胞之能力仍未進一步評估。本研究使用包括三陰性惡性乳癌細胞與其它具有不同分子表達趨勢之惡性乳癌細胞來評估TP4抗腫瘤能力;並進一步利用基因體學分析TP4可能在乳癌細胞中誘導之潛力治療因子。此外,我們利用斑馬魚異體移植模式進一步探討TP4的治療潛力。
經由細胞存活分析實驗顯示,人工合成之TP4能夠選擇性的毒殺惡性乳癌細胞株 (MDA-MB-231/MDA-MB-453/MCF-7) 而對於正常細胞 (M10/HDF) 的毒性相對輕微。利用基因微陣列分析 (Microarray analysis) 發現,TP4可以調控MDA-MB-231細胞內FOS家族轉錄因子表現量。使用細胞轉染實驗證實大量表達FOS家族轉錄因子之一的FOS可以引導惡性乳癌細胞死亡。此外,我們發現MDA-MB-231異體移植對斑馬魚造成顯著死亡;而在不影響斑馬魚胚胎發育,且不傷害魚體表皮及較不影響生存的安全劑量下,我們發現TP4可以延長MDA-MB-231異體移植斑馬魚之壽命。 本論文證實TP4對於三陰性乳癌細胞具選擇性毒殺能力並具有治療三陰性乳癌之潛力。此外FOSB亦可作為有效治療藥物的預後因子。 | zh_TW |
dc.description.abstract | Among the breast cancer subtypes, triple negative breast cancer (TNBC) (ER/PR/HER2) is particularly challenging and difficult-to-treat. Combinations of surgery, hormone, irradiation, and targeted therapies are not usually effective to TNBC patients but chemotherapy was shown good therapeutic efficacy. However, failure in TNBCs treatment by chemotherapy is the cause of multidrug-resistant (MDR) cancer cell developed and healthy cells are disrupted, causing adverse side-effects. Therefore, identifying the molecular targets underlying chemotherapy could contribute to the development of novel cytotoxic agents as well as the improvement of TNBC killing selectivity. Cationic antimicrobial peptides (CAPs) are cytotoxic to certain cancer types, but are less toxic to normal cells. Nile Tilapia piscidin 4 (TP4) is a CAP derived from Nile tilapia (Oreochromis niloticus). Synthetic TP4 was shown to inhibit gram-positive/negative bacteria; however, its role in BC toxicity has not been investigated. In this study, three BC cell lines were used for evaluating the therapeutic efficacy of TP4 in vitro. Transcriptome study was carried out to explore the potential therapeutic factors induced by TP4 in TNBC cell. In addition, zebrafish xenograft model was developed to investigate the therapeutic efficacy of TP4 in vivo.
Cell viability assay showed that synthetic TP4 selectively damages to BC cells (MDA-MB-231/MDA-MB-453/MCF-7) but is less toxic to normal cells (M10/HDF). Microarray study identified FOS family proteins were affected by TP4 treatment in TNBC cells and that overexpression of a FOS family protein, FOSB, caused dramatically TNBC cell death. In addition, we observed TNBC xenotransplantation caused significant zebrafish death in vivo; however, TP4 prolongs the survival in TNBC xenograft zebrafish under a safe dosage. In this study, we confirmed that synthetic TP4 shows selective cytotoxicity to TNBC cell and good therapeutic efficacy. We also identified FOSB as a probable prognostic factor among effective TNBC treatment. | en |
dc.description.provenance | Made available in DSpace on 2021-06-08T00:49:23Z (GMT). No. of bitstreams: 1 ntu-104-R02B45010-1.pdf: 2944820 bytes, checksum: 774a7e34366f4cc651adbf058d5d0a7d (MD5) Previous issue date: 2015 | en |
dc.description.tableofcontents | 目錄 i
摘要 iv Abstract v 第一章 前言 1 1.1乳癌 (Breast cancer) 1 1.2乳癌的異質性 (Heterogeneity) 1 1.3乳癌治療方法 2 1.3.1外科手術 (Surgery) 2 1.3.2放射線治療 (Radiation therapy) 2 1.3.3化學治療 (Chemotherapy) 2 1.3.4賀爾蒙治療 (Hormone therapy) 3 1.3.5標靶治療 (Molecular targeted therapy) 3 1.4三陰性乳癌細胞 (Triple negative breast cancer/TNBC) 3 1.5 Activator protein-1 (AP-1)家族表現影響乳癌細胞生長 4 1.6抗菌胜肽 (Anti-Microbial Peptides/AMP) 4 1.7抗菌胜肽之活性 5 (a)桶狀穿孔模式 (The barrel-stave model) 5 (b)螺旋穿孔模式 (The toroidal model) 5 (c)穿透模式 (The carpet model) 5 1.8 Tilapia piscidin 4 (TP4) 6 1.9抗菌胜肽的抗癌潛力 6 1.10斑馬魚異體移植模式 (Zebrafish xenograft model) 6 1.10.1斑馬魚異體移植模式的優劣 7 1.11斑馬魚的免疫調控 8 1.12研究目的 9 第二章、實驗材料與方法 10 2.1 實驗材料 10 2.1.1 細胞株 10 2.1.2 實驗動物 10 2.1.3 Tilapia piscidin 4 (TP4) 人工合成 10 2.1.4 去氧核醣核酸質體 (plasmid DNA) 11 2.1.5 專一性引子 (primer) 11 2.1.6 試劑 12 2.1.7 藥品 13 2.1.8 耗材 14 2.1.9 抗體 14 2.1.11 儀器設備 15 2.1.12 使用軟體 16 2.2 實驗方法 17 2.2.1 細胞培養 (Cell culture) 17 2.2.2 細胞存活分析 (Cell viability assay/MTS assay) 17 2.2.3 核糖核酸萃取與純化 (RNA extraction and purification) 17 2.2.4 基因微陣列分析 (Microarray study) 18 2.2.5 西方墨點法 (Western Blot) 18 2.2.6 質體DNA的轉染與穩定細胞株篩選 (Transfection and stable clone selection) 19 2.2.7 去氧核醣核酸片段化分析 (DNA laddering analysis) 19 2.2.8 乳酸脫氫酶測定法 (Lactate dehydrogenase analysis/LDH analysis) 20 2.2.9 斑馬魚異體移植分析 (Zebrafish xenograft assay) 20 2.2.10 TP4在斑馬魚之毒性測試 (TP4 toxicity evaluation in zebrafish) 20 2.2.11 斑馬魚固定與高內涵影像分析 (Zebrafish immobilization and High content imaging) 20 2.2.12 斑馬魚免疫螢光染色與共軛焦顯微鏡分析 (Whole-mount immunohistochemical staining and confocal microscopic studies) 21 2.2.13 斑馬魚核醣核酸萃取與純化 (Zebrafish RNA extraction and purification) 22 2.2.14 反轉錄作用與定量聚合酶鏈反應 (Reverse transcription and quantitative PCR) 22 2.2.15 統計分析: 23 第三章、實驗結果 24 3.1 TP4對於乳癌細胞株具有選擇性的毒殺效力 24 3.2 基因微陣列資訊分析顯示FOS家族轉錄因子會受到TP4之誘導 25 3.3 惡性乳癌細胞株存活率會受到FOSB轉錄因子的影響 25 3.4 TP4造成MDA-MB-231細胞死亡之機制 26 3.5 MDA-MB-231異體移植對斑馬魚幼魚存活率之影響 26 3.6 TP4治療能夠延長MDA-MB-231異體移植斑馬魚壽命 27 3.7探討TP4在異體移植斑馬魚模式造成腫瘤細胞自主性與非自主性之治療效益 28 3.8 MDA-MB-231異體移植造成斑馬魚死亡之機制 29 第四章 討論 30 4.1 TP4造成乳癌細死亡之機制探討 30 4.2 FOS家族轉錄因子的表現與乳癌細胞死亡之機制 31 4.3三陰性乳癌細胞造成斑馬魚死亡之機制 32 4.4 TP4在三陰性乳癌異體移植斑馬魚之治療機制 33 (1) TP4誘導異體移植斑馬魚體內的MDA-MB-231細胞自主性毒殺作用 33 (2) TP4可調節異體移植斑馬魚之先天免疫反應 34 第五章 結論與展望 35 第六章 參考文獻 36 圖 42 附錄 68 | |
dc.language.iso | zh-TW | |
dc.title | 建立斑馬魚異體移植模式探討陽離子抗菌蛋白TP4治療三陰性乳癌的功效 | zh_TW |
dc.title | Generation of a zebrafish xenograft model of triple-negative breast cancer for evaluating the therapeutic efficacy of Tilapia piscidin 4 (TP4) | en |
dc.type | Thesis | |
dc.date.schoolyear | 103-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 吳金洌(Jen-Leih Wu),潘婕玉(Chieh-Yu Pan) | |
dc.subject.keyword | TP4,三陰性乳癌,FOSB,異體移植斑馬魚, | zh_TW |
dc.subject.keyword | TP4,triple negative breast cancer (TNBC),FOSB,xenograft zebrafish, | en |
dc.relation.page | 69 | |
dc.rights.note | 未授權 | |
dc.date.accepted | 2015-07-13 | |
dc.contributor.author-college | 生命科學院 | zh_TW |
dc.contributor.author-dept | 漁業科學研究所 | zh_TW |
顯示於系所單位: | 漁業科學研究所 |
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