臺灣博碩士論文加值系統

中文摘要 I
Abstract II
縮寫檢索表 IV
第一章、緒論 1
第二章、文獻探討 2
2.1 頭頸部癌(Head and neck cancer) 2
2.1.1 簡介 2
2.1.2臨床治療及現況 2
2.1.3 頭頸部癌與多酚類(polyphenol)物質之相關性 2
2.2 順鉑(cisplatin) 3
2.2.1 cisplatin藥物簡介 3
2.2.2 cisplatin合併多酚類物質輔助癌症治療之相關研究 5
2.3蓮蓬(Lotus seedpod) 5
2.3.1 蓮植株介紹 5
2.3.2蓮蓬成分與功效 6
2.4細胞凋亡(Apoptosis) 6
2.4.1 細胞凋亡定義 6
2.4.2細胞凋亡的型態與特徵 7
2.4.3 細胞凋亡分子機制 7
2.5 細胞自噬(Autophagy) 8
2.5.1 細胞自噬簡介 8
2.5.2 細胞自噬與疾病之關係 9
2.5.3 細胞自噬分子機制 10
第三章、實驗架構與流程 12
3.1細胞與動物實驗架構 12
第四章、材料與方法 13
4.1實驗儀器與藥品試劑 13
4.1.1 各項實驗儀器與試劑 13
4.1.2 Cisplatin之配製 20
4.1.3 蓮蓬萃取物(lotus seedpod extracts, LSE)之製備 20
4.2細胞培養(Cell culture) 20
4.2.1 細胞株 20
4.2.2 實驗設計 20
4.2.3 培養條件 20
4.2.4 解凍細胞 21
4.2.5 繼代培養 21
4.2.6 細胞計數 21
4.2.7 冷凍細胞 21
4.3 細胞存活試驗(Cell viability) 22
4.4細胞凋亡分析(螢光染色法4’6-diamidino-2-phenylindole, DAPI stain) 22
4.5細胞凋亡分析-雙染色法(Annexin V/PI stain) 22
4.6 凋亡蛋白酶活性測試(cysteine aspartate protease activity assay) 23
4.7.細胞蛋白質萃取與定量 23
4.7.1. 蛋白質萃取 23
4.7.2 蛋白質定量 24
4.7.3 粒線體分離 24
4.8. 西方墨點法(Western blotting) 24
4.8.1 樣品(Sample)配置 24
4.8.2 架膠 (SDS-PAGE) 25
4.8.3 電泳法(electrophoresis) 25
4.8.4 轉漬(Transfer) 26
4.8.5 免疫墨點法(Immunoblot) 26
4.9 粒線體膜電位分析(Mitochondria membrane potentialassay, JC-1 staining) 27
4.10 細胞自噬分析(Acridine orange stain, AVO stain) 27
4.11 實驗動物 28
4.11.1 實驗設計與流程 28
4.11.2 飼料配製 28
4.11.3 Cisplatin 注射液配置 29
4.12.4 樣本採集與處理 29
4.12 統計方法 29
第五章、實驗結果 30
5.1探討不同濃度LSE單獨及合併cisplatin處理對於頭頸部癌FaDu細胞存活之影響 30
5.2探討LSE合併cisplatin處理對於FaDu細胞凋亡之影響 30
5.3 探討LSE合併cisplatin處理對於FaDu細胞內凋亡蛋白酶活性之影響 31
5.4 探討LSE合併cisplatin處理對於FaDu細胞內凋亡分子表現之影響 31
5.5 探討LSE合併cisplatin處理對於FaDu 細胞內粒線體膜電位改變之影響 31
5.6 探討LSE合併cisplatin處理對於調控凋亡內在因子cytochrome c表現量之影響 32
5.7探討LSE合併cisplatin處理對於調控凋亡內在因子Bcl-family蛋白之影響 32
5.8 探討LSE合併cisplatin處理對於自噬細胞之影響 33
5.9 探討LSE合併cisplatin處理對於細胞自噬相關蛋白之影響 33
5.10 探討預先處理細胞自噬抑制劑對於LSE合併cisplatin之細胞凋亡的影響 34
5.11 探討預先處理細胞自噬抑制劑對於LSE合併cisplatin之細胞自噬的影響 34
5.12 探討LSE合併cisplatin處理對於裸鼠移植性腫瘤大小以及生長速度之影響 35
5.13 探討LSE合併cisplatin處理對於體內腫瘤組織細胞凋亡相關蛋白表現之影響 35
5.14 探討LSE合併cisplatin處理對於體內腫瘤組織細胞自噬相關蛋白表現之影響 36
第六章、討論 37
第七章、結論 40
第八章、參考文獻 41

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