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dc.contributor.authorLiu, Yi-Shianen_US
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dc.description.abstract空氣懸浮微粒是具有健康危害效應的物質。文獻指出,空氣懸浮微粒上所吸附的脂溶性、水溶性及其他物質,是造成細胞胞內氧化壓力累積生成的重要因素之一。此外文獻亦提到,氧化壓力會造成人體老化、內分泌失調以及癌症等疾病。因此探討空氣懸浮微粒形成氧化壓力以及細胞毒性的機制,可以協助了解空氣懸浮微粒致細胞毒性及致癌作用機轉。本研究目的主要是去探討,曝露在空氣懸浮微粒之水溶性萃取樣本下,細胞內部是否為經由代謝活化效應誘發基因表現失衡,進而造成細胞胞內氧化壓力、DNA損害以及細胞死亡。本研究利用多種生物性指標,包括細胞存活率測試(SRB assay)、氧化壓力測定法(Fluorescence assay)、DNA單股斷裂分析法(COMET assay)以及西方墨點法(Western blot),針對懸浮微粒可能產生之細胞毒性、氧化壓力、基因損害以及基因表現進行研究與分析。根據實驗結果顯示,曝露在空氣懸浮微粒水溶性萃取物質下,不論是MDA-MB-231細胞還是MCF-7細胞均無明顯的細胞毒性效應生成。研究結果也顯示出,曝露於空氣懸浮微粒水溶性萃取物2小時下即有氧化壓力顯著上升。在懸浮微粒誘發DNA單股損害部分,三個採樣點的水溶性萃取液均能造成兩種乳癌細胞顯著之DNA單股斷裂。此外針對細胞之基因表現部分,MCF-7細胞曝露在東大以及草屯採樣站脂溶性樣本萃取物下,其p53蛋白有明顯上升;曝露於三個採樣站之脂溶性樣本則會造成CYP1A1代謝酵素明顯增加。綜合本研究之結果可知,空氣懸浮微粒的水溶性萃取物質,可以造成人類乳癌細胞胞內氧化壓力上升及基因單股斷裂。而脂溶性萃取物質則會造成MCF-7細胞p53以及CYP1A1基因大量表現,進而造成細胞凋亡以及胞內氧化壓力之生成。zh_TW
dc.description.tableofcontents摘要 I Abstract II Abbreviation IV 目錄 VI 圖目錄 IX 表目錄 XI 第一章 前言 1 1-1研究緣起 1 1-2研究目的 1 第二章 文獻回顧 3 2-1 懸浮微粒(PM) 3 2-1-1 空氣懸浮微粒之分類 3 2-1-2 PM的性質以及環境中分布的比例 4 2-1-3 懸浮微粒的來源、成分以及其健康危害 6 2-2 空氣懸浮微粒(PM)所誘發之健康危害 10 2-2-1 環境污染物對呼吸系統之影響 10 2-2-2 PM對人體健康之流行病學資料 11 2-2-3 PM對人類呼吸系統之影響 11 2-2-4 PM對人類乳癌及生殖系統之影響 12 2-2-5 PM對人類心血管疾病之影響 12 2-3 懸浮微粒(PM)對人類健康之危害效應及生物體之影響 12 2-3-1 PM誘發之細胞毒性(Cytotoxicity) 12 2-3-2 PM誘發之活性氧分子(Reactive oxygen species, ROS) 14 2-3-3 PM誘發之DNA 損壞(DNA damage) 16 2-3-4 PM致突變能力(Mutagenicity) 18 2-3-5 PM粒徑差異對健康之危害效應 18 2-3-6 PM之季節差異對健康之危害 18 第三章 實驗材料與方法 19 3-1 實驗材料 19 3-1-1 化學藥品 19 3-1-2 人類細胞株來源 20 3-1-3 實驗設備 20 3-2 實驗方法 20 3-2-1 空氣懸浮微粒樣本收集 20 3-2-2 濾紙樣本萃取及生物檢測前處裡 22 3-2-3 細胞培養 23 3-2-4 細胞毒性測試 24 3-2-5 細胞存活率測試(Sulforhodamine B assay, SRB) 24 3-2-6 螢光分析活潑性氧分子生成量(Fluorescence assay) 25 3-2-7 彗星試驗(Comet assay) 25 3-2-8 西方墨點法(Western Blot) 29 3-2-9 水溶性萃取樣本成分分析 30 3-2-10 數據分析 30 第四章 實驗架構 31 4-1 實驗架構 31 4-1-1懸浮微粒水溶性萃取物,對誘發人類乳癌細胞之細胞毒性、胞內氧化壓力、核酸損壞與基因表現之影響 31 4-1-2懸浮微粒脂溶性萃取物,對人類乳癌細胞誘發細胞凋亡、CYP1A1代謝酵素以及核酸修補之基因表現 32 4-1-3不同粒徑範圍懸浮微粒脂溶性萃取物質,對誘發人類乳癌細胞之細胞毒性、胞內氧化壓力及核酸氧化損害之影響 33 第五章 實驗結果 34 5-1 總懸浮微粒(TSP)之細胞實驗結果 34 5-1-1 總懸浮微粒(TSP)水溶性萃取液之細胞毒性試驗 (Sulforhodamine B assay, SRB assay) 34 5-1-1-1總懸浮微粒(TSP)水溶性萃取液誘發細胞毒性 34 5-1-1-2 TSP水溶性萃取液之暴露濃度(concentrate-dependent) 及暴露時間(time-dependent)對於MDA-MB-231細胞之毒性測試之結果 34 5-1-1-3 TSP水溶性萃取液之暴露濃度(concentrate-dependent) 及暴露時間(time-dependent)對於MCF-7細胞之毒性測試之結果 35 5-1-1-4 TSP水溶性萃取樣本對兩種細胞之細胞毒性之影響 35 5-1-2 螢光分析活潑性氧分子生成量(Fluorescence assay) 36 5-1-2-1 TSP水溶性萃取液之暴露濃度(concentrate-dependent) 及暴露時間(time-dependent)對於MDA-MB-231細胞生成ROS之影響 36 5-1-2-2 TSP水溶性萃取液之暴露濃度(concentrate- dependent)及暴露時間(time-dependent)對於MCF-7細胞生成ROS之影響 36 5-1-3 TSP 水溶性萃取物對細胞DNA 之單股斷鏈作用(Comet assay) 37 5-1-3-1 TSP 水溶性萃取物於MDA-MB-231 細胞DNA 單股斷鏈作用 37 5-1-3-2 TSP 水溶性萃取物於MCF-7 細胞DNA 單股斷鏈作用 37 5-1-4 西方墨點法(Western blot assay) 38 5-1-4-1 TSP 脂溶性萃取物對於細胞毒性之影響 38 5-1-4-2 TSP 脂溶性萃取物對於代謝酵素CYP1A1 之影響 38 5-1-4-3 TSP 脂溶性萃取物對基因修補酵素之影響 38 5-1-4-4 TSP 水溶性萃取物對於細胞毒性之影響 39 5-1-4-5 TSP 水溶性萃取物對於代謝酵素CYP1A1 之影響 39 5-2 TSP、PM10 及PM2.5 細胞毒性、氧化壓力及基因損害之結果 70 5-2-1 不同粒徑相同重量下之細胞毒性效應 70 5-2-2 不同粒徑相同重量下之氧化壓力生成效應 70 5-3 TSP 金屬成分分析之結果 73 5-3-1 不同時間地點之TSP 金屬成分分析 73 第六章 討論 74 6-1 懸浮微粒細胞毒性之誘發(cytotoxicity) 74 6-2 懸浮微粒氧化壓力之生成(Oxidative stress) 78 6-3 懸浮微粒誘發基因單股斷裂(Single strand break) 81 6-4 懸浮微粒誘發乳癌細胞之基因表現 81 6-4-1 脂溶性萃取物誘發乳癌細胞之基因表現 81 6-4-2 水溶性萃取物誘發乳癌細胞之基因表現 82 6-5 不同粒徑懸浮微粒之比較 82 6-5-1 不同粒徑對細胞毒性之誘發(Cytotoxicity) 82 6-5-2 不同粒徑對細胞氧化壓力之誘發 82 6-6 懸浮微粒金屬成分分析 83 6-6-1 TSP 濾紙之金屬成分分析 83 6-7 結論 84 第七章 未來建議 88 第八章 參考文獻 89zh_TW
dc.subjectParticulate matteren_US
dc.subjectOxidative stressen_US
dc.subjectGene expressionen_US
dc.titleInduction of cytotoxic response and oxidative DNA damage in human breast cancer cell lines by water- and solvent-extractable particulate matter in central Taiwanen_US
dc.typeThesis and Dissertationzh_TW
item.openairetypeThesis and Dissertation-
item.fulltextno fulltext-
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