Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/5505
DC FieldValueLanguage
dc.contributor林嬪嬪zh_TW
dc.contributor王應然zh_TW
dc.contributor.advisor林伯雄zh_TW
dc.contributor.author劉懿賢zh_TW
dc.contributor.authorLiu, Yi-Shianen_US
dc.contributor.other中興大學zh_TW
dc.date2009zh_TW
dc.date.accessioned2014-06-06T06:34:55Z-
dc.date.available2014-06-06T06:34:55Z-
dc.identifierU0005-2508200814241100zh_TW
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dc.identifier.urihttp://hdl.handle.net/11455/5505-
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.language.isoen_USzh_TW
dc.publisher環境工程學系所zh_TW
dc.relation.urihttp://www.airitilibrary.com/Publication/alDetailedMesh1?DocID=U0005-2508200814241100en_US
dc.subjectParticulate matteren_US
dc.subject懸浮微粒zh_TW
dc.subjectOxidative stressen_US
dc.subjectGene expressionen_US
dc.subject氧化壓力zh_TW
dc.subject基因表現zh_TW
dc.title台灣中部地區空氣懸浮微粒之水溶性及脂溶性萃取物誘發人類乳癌細胞之細胞毒性和DNA氧化損害zh_TW
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.openairecristypehttp://purl.org/coar/resource_type/c_18cf-
item.openairetypeThesis and Dissertation-
item.cerifentitytypePublications-
item.fulltextno fulltext-
item.languageiso639-1en_US-
item.grantfulltextnone-
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