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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
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dc.contributor.advisor | 繆希椿(Shi-Chuen Miaw) | |
dc.contributor.author | Min Chen | en |
dc.contributor.author | 陳名 | zh_TW |
dc.date.accessioned | 2021-06-17T06:36:12Z | - |
dc.date.available | 2028-08-15 | |
dc.date.copyright | 2018-09-04 | |
dc.date.issued | 2018 | |
dc.date.submitted | 2018-08-16 | |
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/72337 | - |
dc.description.abstract | 目前已知癌症細胞分泌出的胞外小體(exosomes)會幫助血管新生、凝血機制、調控免疫系統,以及改造週邊的薄壁組織(parenchymal tissue),綜合上述功能,這些胞外小體就具有幫助癌症發展的能力。故在此研究中,我們要探討同為黑色素瘤(melanoma)細胞株的B16-F1以及B16-F10兩者之間的癌症轉移能力是否與胞外小體內容物有關。另一方面,TH17細胞的主要功能為引導更多的免疫細胞到癌症細胞或是病原體所在處,以及事後的組織修復。然而,過去研究已知TH17細胞的可塑性(plasticity)可使其轉變(reprogramming)成其他的輔助型T細胞,甚至能轉變成幫助癌症生長的Treg細胞。因此,進一步我們要證實由B16-F1以及B16-F10產生的胞外小體是否導致TH17細胞轉變成Treg細胞。在本研究中,我們首先分離出B16-F1以及B16-F10產生的胞外小體,並且確認他們的大小和表面蛋白符合胞外小體。再進一步,我們將分離出的B16-F1以及B16-F10胞外小體加入正在分化為TH17細胞的naïve T細胞中,我們發現兩種來源的exosomes隨著施加的exosomes蛋白濃度提高至20 g/ml,皆不影響TH17細胞的可塑性。 | zh_TW |
dc.description.abstract | Tumor-derived exosomes can promote angiogenesis and coagulation, modulate the immune system, and remodel surrounding parenchymal tissue, which together support tumor progression. Therefore, we aimed to characterize the content within the tumor-derived exosomes from B16-F1 and B16-F10, which are known to have different metastasis potential. Furthermore, TH17 cells are responsible for recruiting immune cells against cancer cells or pathogens, and tissue repairing after pathogen clearance. However, TH17 cells are known to have the plasticity to reprogram into other T helper subtypes, for example, Treg cells, which are pro-tumor growth cells. Therefore, we further examined whether B16-F1/F10 exosomes reprogram TH17 cells into Treg cells. Hence, we first isolated and characterized the size and surface proteins of exosomes by Nanoparticle tracking analysis (NTA) and Western blot, respectively. Furthermore, we cultured TH17 cells with isolated B16-F1- and B16-F10-derived exosomes, and identified the TH17 plasticity by quantitative PCR analysis. The results showed that both B16-F1- and B16-F10-derived exosomes do not affect the expression of TH17 related genes with the concentration of exosome protein up to 20 μg/ml. | en |
dc.description.provenance | Made available in DSpace on 2021-06-17T06:36:12Z (GMT). No. of bitstreams: 1 ntu-107-R05449011-1.pdf: 2013537 bytes, checksum: cce48512248c096fdd7fd94408a47585 (MD5) Previous issue date: 2018 | en |
dc.description.tableofcontents | 致謝 I
中文摘要 II 英文摘要 III 目錄 IV 第一章、緒論 1 1.1 癌症轉移 1 1.2 黑色素瘤 1 1.2.1 目前針對黑色素瘤的治療方法 2 1.3 過繼性細胞免疫療法 (adoptive cell transfer, ACT) 2 1.4 CD4+輔助型T細胞 3 1.4.1 TH1輔助細胞 3 1.4.2 TH2輔助細胞 4 1.4.3 TH17輔助細胞 4 1.4.4 調節型T細胞(Treg) 5 1.5 TH17細胞在過繼性細胞免疫療法(ACT)的角色 6 1.6 胞外小體(Exosomes) 7 第二章、材料與方法 10 一、 實驗材料 10 1.1 實驗小鼠(Mice) 10 1.2 抗體和細胞激素(Antibodies and cytokines) 10 1.3 緩衝液和培養液配方(Buffers and Mediums) 12 1.4 化學藥品及試劑 (Chemicals and Reagents) 18 1.5 引子 (Primers) 20 二、 實驗方法 21 1.1 B16-F1和B16-F10細胞株的培養 21 1.2 胞外小體的純化 (exosome isolation) 21 1.3 奈米粒子追蹤分析儀 (nanoparticle tracking analysis, NTA) 22 1.4 西方墨點法 (Western blot) 23 1.5 流式細胞分選 (cell sorting) 23 1.6 流式細胞分析 (flow cytometry) 24 1.7 定量即時聚合酶連鎖分析 (quantitative real-time polymerase chain reaction, RT-QPCR) 24 1.8 酵素結合免疫吸附分析法 (enzyme-linked immunosorbent assay, ELISA) 24 1.9 共同培養輔助型TH17細胞與B16-F1或B16-F10胞外小體 25 1.10 黴漿菌 (mycoplasma) PCR 檢測 25 1.11 統計分析 26 第三章、實驗結果 27 1. B16-F1和B16-F10細胞與未處理的胎牛血清或剔除胎牛胞外小體的胎牛血清培養後的存活率並無顯著差異。 27 2. B16-F1、B16-F10以及細胞培養液DMEM沒有黴漿菌的汙染。 27 3. 檢驗B16-F1和B16-F10衍生的胞外小體。 27 4. 以ELISA和流式細胞儀確認TH17細胞的分化。 28 5. B16-F1和B16-F10的胞外小體濃度在高達20 μg/ml,仍無影響TH17細胞的可塑性。 29 6. B16-F1和B16-F10的胞外小體濃度在高達60 μg/ml,仍無影響TH0細胞的分化。 30 第四章、實驗討論 31 1. 黑色素瘤細胞衍生的胞外小體可能不影響TH17的可塑性和TH0細胞的分化。 31 2. 黑色素瘤細胞衍生的胞外小體內容物檢驗。 31 3. 黑色素瘤細胞衍生的胞外小體產量低。 32 4. 可更精準的分離出黑色素瘤細胞衍生的胞外小體。 33 第五章、圖表 34 圖一、B16-F1和B16-F10細胞與未處理的FBS或剔除胎牛胞外小體的FBS培養後的存活率並無顯著差異。 36 圖二、B16-F1、B16-F10以及細胞培養液DMEM沒有黴漿菌的汙染。 38 圖三、檢驗B16-F1和B16-F10衍生的胞外小體。 42 圖四、以ELISA和流式細胞儀確認TH17細胞的分化。 45 圖五、B16-F1和B16-F10的胞外小體濃度在高達20 μg/ml,仍無影響TH17細胞的可塑性。 47 圖六、B16-F1和B16-F10的胞外小體濃度在高達60 μg/ml,仍無影響TH0細胞的分化。 49 第六章、參考文獻 50 | |
dc.language.iso | zh-TW | |
dc.title | 癌症衍生之胞外小體對於TH17細胞的作用 | zh_TW |
dc.title | The effect of Tumor-derived Exosomes on TH17 cells | en |
dc.type | Thesis | |
dc.date.schoolyear | 106-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 李建國(Chien-Kuo Lee),賴逸儒 | |
dc.subject.keyword | 癌症衍生之胞外小體,黑色素瘤,癌症轉移,TH17輔助型細胞,可塑性, | zh_TW |
dc.subject.keyword | Tumor-derived exosomes,melanoma,metastasis,TH17,plasticity, | en |
dc.relation.page | 60 | |
dc.identifier.doi | 10.6342/NTU201803397 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2018-08-16 | |
dc.contributor.author-college | 醫學院 | zh_TW |
dc.contributor.author-dept | 免疫學研究所 | zh_TW |
顯示於系所單位: | 免疫學研究所 |
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