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姓名	林政賢(Cheng-Hsien Lin)  查詢紙本館藏	畢業系所	化學工程與材料工程學系
論文名稱	利用聚己內酯/褐藻酸鈉之複合電紡絲擴增癌症幹細胞 (Electrospinning of Polycaprolactone/Alginate Composite Nanofibers for Cancer Stem Cell Enrichment)
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摘要(中)	雖然褐藻酸鈉製成的電紡絲支架可以用來篩選出具有較佳抗藥性的癌細胞，但是其細胞貼附率和細胞增生的能力都較差，因此我們加入聚己內酯纖維進行混合紡絲。透過改變高分子進料的流率，我們得以控制複合纖維的組成。之後將複合纖維篩選出的癌細胞進行抗藥性實驗，發現與褐藻酸鈉沒有明顯的差別，但是在細胞的貼附與增生上有上升的現象。為了瞭解篩選出來之癌細胞之細胞特性，我們利用qPCR去分析細胞的基因表現，發現褐藻酸鈉和複合材料篩選出的細胞具有較佳的幹性、侵入性、化學抗性、上皮間質轉換和血管新生的基因表現，代表我們所篩選出來的是癌症幹細胞。透過傷口癒合實驗、群落形成實驗和細胞侵入性實驗發現，利用褐藻酸鈉和複合材料篩選出來的癌症幹細胞表現都比聚己內酯纖維來的好，此結果與qPCR基因表現可相互驗証。這些結論都顯示了利用複合纖維支架來進行癌症幹細胞的篩選，在癌症藥物的研究和治療上都可以有很大的助益。
摘要(英)	Although electrospun alginate scaffolds can be used to enrich cancer cells with high drug resistance, the low cell adhesion and proliferation restricts its application. Therefore, we incorporated polycaprolactone (PCL) for coelectrospinning to prepare composite nanofibrous scaffolds. The composition of nanofibers can be regulated through adjusting the perfusion rates of polymer jets. These composite scaffolds were applied to culture cancer cell lines. The enriched cells were treated anti-cancer drug to evaluate their drug resistances. The results suggested that the drug resistance of cells selected from composite scaffolds was similar to those from alginate fibers. However, their adhesion and proliferation were highly improved. We applied qPCR to analyzed gene expression of selected cells, and the results demonstrated that cell collected from alginate and composite fibers all exhibited high stemness, invasion, chemical resistance, epithelial-mesenchymal transition, and angiogenesis compared to those on PCL fibers, suggesting these cells should be cancer stem cells. Furthermore, the data of wound healing, colony forming, and cell invasion experiments were also consistence to the qPCR results that alginate and composite fibers can enrich cells exhibiting high cancer-stem-cell properties. These results indicated that cancer stem cell enrichment through composite nanofibrous scaffolds should be beneficial to anti-cancer drug development and the research of cancer therapy.
關鍵字(中)	★ 奈米電紡絲 ★ 癌症幹細胞 ★ 擴增	關鍵字(英)	★ Electrospun nanofiber ★ Cancer stem cell ★ Enrichment
論文目次	摘要 i Abstract ii 致謝 iii 目錄 viii 圖目錄 xi 表目錄 xiv 第一章 緒論 1 1.1 背景 1 1.2 實驗目的 2 第二章 文獻回顧 3 2.1 癌症 3 2.1.1 癌症幹細胞 3 2.1.2 癌症幹細胞特性 5 2.1.3 癌症幹細胞篩選 6 2.1.4 癌症幹細胞相關基因 9 2.2 電紡絲 13 2.2.1 電紡絲原理 13 2.2.2 電紡絲於癌症幹細胞的應用 15 2.3 褐藻酸鈉 18 2.3.1 褐藻酸鈉之來源 18 2.3.2 褐藻酸鈉之性質 18 2.3.3 褐藻酸鈉於癌症幹細胞的應用 19 2.4 聚己內酯 22 2.4.1 聚己內酯之來源 22 2.4.2 聚己內酯之性質 22 2.4.3 聚己內酯於癌症幹細胞的應用 23 2.5 複合材料 25 2.5.1 複合材料之簡介 25 2.5.2 複合材料於癌症幹細胞的應用 25 2.6電紡絲製備複合材料 28 2.6.1 電紡絲製備複合材料的應用 29 第三章 實驗與方法 31 3.1 實驗藥品 31 3.2 實驗儀器 35 3.3 實驗方法 37 3.3.1 電紡絲溶液製備 37 3.3.2 電紡絲纖維製備 38 3.3.3 收集量量測 38 3.3.4 纖維材料表面分析 39 3.3.5 生物實驗 39 第四章 結果與討論 54 4.1 褐藻酸鈉/聚己內酯電紡絲抗藥性 54 4.1.1 篩選之細胞於支架外抗藥性 54 4.1.2 支架上之抗藥性表現 60 4.2 褐藻酸鈉/聚己內酯之複合纖維性質 65 4.2.1 纖維之收集量 65 4.2.2 複合纖維之收集 69 4.3 支架對細胞形態的影響 70 4.3.1 細胞於纖維上之螢光圖 70 4.3.2細胞於纖維上之SEM圖 75 4.4 複合材料之藥物抗性 77 4.5 癌症幹細胞相關基因表現 82 4.5.1 癌細胞之幹性相關基因表現 82 4.5.2 癌症幹細胞之上皮間質轉換相關基因 85 4.5.3 癌症幹細胞之侵入性相關基因 88 4.5.4 癌症幹細胞之血管新生相關基因 90 4.5.5 癌症幹細胞之化學抗性相關基因 92 4.6 癌症幹細胞之群落形成 94 4.7 癌症幹細胞之細胞移動能力 98 4.8 癌症幹細胞之細胞侵入能力 103 第五章 結論 105 第六章 參考資料 107
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指導教授	胡威文(Wei-Wen Hu)	審核日期	2016-8-30
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