Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/5600
DC FieldValueLanguage
dc.contributor.advisor林伯雄zh_TW
dc.contributor.advisorPo-Hsiung Linen_US
dc.contributor.author潘文驥zh_TW
dc.contributor.authorPan, Wen-Chien_US
dc.date2003zh_TW
dc.date.accessioned2014-06-06T06:35:09Z-
dc.date.available2014-06-06T06:35:09Z-
dc.identifier.urihttp://hdl.handle.net/11455/5600-
dc.description.abstract本研究之主要目的為探討萘之醌類代謝物1,2-dihydroxynaphthalene(NCAT)及1,4-dihydroxynaphthalene(NHQ)能否經由細胞內之氧化還原循環,進而生成活性氧化物(reactive oxygen species, ROS)誘發人類乳癌細胞T-47D核酸醛基損害(aldehydic DNA lesion, ADL)及其生成之機制。研究結果顯示,NCAT可以誘發人類乳癌細胞T-47D ADL 之生成,且與細胞反應1.5小時後ADL生成量與控制組相較達到最大值(p<0.001),但延長反應時間(至8 — 16 小時),此一細胞核酸損壞作用即回覆正常(p>0.05)。但測試細胞移除NCAT所誘發之ADL之DNA修補效率實驗發現,即是經更新培養液之細胞,經過6.5小時再培養(post-incubation),其所誘發之ADL依舊存在(p<0.05)。此一研究結果顯示,可能為NCAT誘發氧化壓力進而促進核酸修補酵素之基因表現所致。若比較NCAT及NHQ誘發細胞之ADL能力的差異,研究結果顯示,NCAT (750μM及1.25 mM)可以誘發ADL之生成且為concentration-dependent,反之,NHQ於相同濃度下並無法有效誘發ADL的生成,但此一誘發細胞核酸損壞作用之研究結果,與SRB細胞存活率測試結果相異,亦即NHQ對人類T47D細胞之毒性較NCAT為強。而利用putrescine-cleavage assay確認由NCAT所誘發的ADL大部分為氧化壓力所致。進一步之研究結果發現,過渡性金屬離子Cu (Ⅰ)、Fe(Ⅱ)、及 過氧化氫參與NCAT誘發ADL的機制。zh_TW
dc.description.abstractThe primary purpose of this research is to examine the potentials of the induction aldehydic DNA lesions (ADL) by quinonoid derivatives of naphthalene, e.g., 1,2-dihydroxynaphthalene (NCAT), 1,4-dihydroxynaphthalene (NHQ) in human T-47D breast cancer cell line. Results indicated that when cells treated with NCAT (1 mM, 2 h), increases in the number of ADL was detected in T-47D cells exposed to NCAT for 1.5 h (p<0.001). When cells exposed to NCAT for additional 6 — 14h, no significant increases in the number of ADL was detected when compared to the corresponding control (p>0.05). In contrast, increases in the number of ADL were detected in cells treated with NCAT for 1.5 h followed by incubation with fresh medium for additional 6.5 h (p<0.05). These findings suggest that NCAT may induce ROS-mediated induction of the expression of BER genes to excise this type of DNA lesions with efficiency in cells. Although NHQ did not induce appreciable increases in the number of ADL when compared to control (p>0.05), NHQ is more toxic than NCAT toward human T47D cells as assayed by SRB assay. Further investigation indicated that the ADL induced by NCAT contain predominantly (100%) putrescine-excisable ADL in T-47D cells. Futher, iron (II) copper (I), hydrogen peroxide participate in the induction of ADL by NCAT in T-47D cells. Overall, this evidence confirmed that the ADL induced by NCAT in T-47D cells is derived from oxidative events.en_US
dc.description.tableofcontents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 多環芳香族碳氫化合物(PAHs)................................3 2-1-1 PAHs之簡介..............................................3 2-1-2 PAHs於環境中之光解作用(photodegradation)與DNA損害.......4 2-1-3 PAH之微生物分解.........................................6 2-1-4 PAHs、芳香烴基受體(aryl hydroarbon receptor, AhR)與雌性激素受體(estrogen receptor, ER).................................7 2-1-5 PAHs醌類衍生物之致基因毒害作用與活性氧化物(ROS)之生成...8 2-2 萘之代謝途徑與致基因毒害作用..............................9 2-2-1 萘之物化特性及應用......................................9 2-2-2 萘於環境中之流佈及人體暴露來源與濃度....................9 2-2-3 萘之健康危害影響.......................................11 2-2-3 萘於哺乳類動物之代謝途徑...............................13 2-2-4 萘之醌類衍生物之致基因毒害作用.........................15 2-3 氧化壓力造成的DNA 損害...................................16 2-3-1 活性氧化物.............................................16 2-3-2 超氧自由基.............................................16 2-3-3 過氧化氫...............................................16 2-3-4 氫氧自由基.............................................17 2-3-5 氧化壓力所誘發之DNA損害................................17 2-3-6 生物體之抗氧化防禦系統.................................18 2-4 DNA損害修補..............................................18 2-4-1 Base Excision Repair (BER).............................19 2-4-2 哺乳類細胞利用BER修補AP-sites及sugar damage之機制......21 第三章 實驗材料與方法........................................22 3-1 實驗材料.................................................22 3-1-1 水.....................................................22 3-1-2 化學藥品...............................................22 3-1-3 細胞培養藥品...........................................22 3-1-4 Sulforhodamine B 分析藥品..............................22 3-1-5 DNA萃取藥品............................................22 3-1-6 核酸醛基損害分析藥品...................................23 3-1-7 細胞凋亡分析法.........................................23 3-1-8 實驗設備...............................................23 3-2 實驗方法.................................................23 3-2-1 細胞培養條件...........................................23 3-2-2 細胞毒性測試(Trypan blue dye exclusion)................24 3-2-3 Sulforhodamine B(SRB) 分析法...........................24 3-2-4 細胞反應條件...........................................25 3-2-4 細胞DNA萃取方法........................................25 3-2-5 核酸醛基損害分析法(aldehyde reactive probe-slot-blot, ASB assay)...................................................26 3-2-6 Putrescine cleavage assay..............................26 3-2-7 細胞凋亡分析法.........................................26 3-3 數據分析.................................................27 第四章 實驗結果..............................................28 4-1 細胞毒性及存活率測試.....................................28 4-2 萘醌誘發DNA醛基損害之能力................................29 4-2-1 1,2-Dihydroxynaphthalene(NCAT)誘發醛基損害的能力.......29 4-2-2 不同之萘醌種類及濃度對誘發醛基損害的能力之影響.........29 4-2-3 DNA修補效率與萘醌誘發之醛基損害之關係..................29 4-3 萘醌誘發DNA醛基損害之作用機制............................30 4-3-1 添加putrescine確認誘發醛基損害之型態...................30 4-3-2 添加Cu (Ⅰ)及Fe(Ⅱ)抑制劑的影響........................30 4-3-3 添加catalase的影響.....................................31 4-3-4 添加ROS移除劑、過渡性金屬螯合劑對細胞存活率之影響......31 4-3-5 細胞凋亡分析...........................................31 第五章 討論..................................................42 第六章 結論與建議............................................49 6-1 結論.....................................................49 6-2 未來研究方向與建議.......................................50 第七章 參考文獻..............................................52zh_TW
dc.language.isoen_USzh_TW
dc.publisher環境工程學系zh_TW
dc.subjectnaphthaleneen_US
dc.subjectzh_TW
dc.subjectADLen_US
dc.subjectcatecholen_US
dc.subjecthydroquinoneen_US
dc.subjectoxidative stressen_US
dc.subject核酸醛基損害zh_TW
dc.subject兒茶酚zh_TW
dc.subject對苯二酚zh_TW
dc.subjectzh_TW
dc.title萘之醌類衍生物誘發人類乳癌細胞T-47D核酸醛基損害之研究zh_TW
dc.titleInduction of aldehydic DNA lesions in human breast cancer cell line T-47D by quinonoid derivitives of naphthaleneen_US
dc.typeThesis and Dissertationzh_TW
item.fulltextno fulltext-
item.languageiso639-1en_US-
item.openairetypeThesis and Dissertation-
item.cerifentitytypePublications-
item.openairecristypehttp://purl.org/coar/resource_type/c_18cf-
item.grantfulltextnone-
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