Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/5080
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dc.contributor.advisor林伯雄zh_TW
dc.contributor.advisorPo-hssiung Linen_US
dc.contributor.author李美質zh_TW
dc.contributor.authorLee, Mei-chihen_US
dc.date2005zh_TW
dc.date.accessioned2014-06-06T06:33:59Z-
dc.date.available2014-06-06T06:33:59Z-
dc.identifier.urihttp://hdl.handle.net/11455/5080-
dc.description.abstract多環芳香族碳氫化合物 (polyaromatic hydrocarbons, PAHs) 為常見的環境污染物質,且為已知之人類之致癌物質。累積研究證據顯示,PAHs亦為荷爾蒙干擾物質,分別具有促進雌性激素 (estrogenic) 及抗拮雌性激素 (anti-estrogenic) 之特性,本研究目的為探討PAHs物質,包括naphthalene (Nap)、7,12-dimethylbenz[a]anthracene (DMBA)、以及鹵化芳香族碳氫化合物,2,3,7,8-tertachlorodibenzo-p-dioxin (TCDD),於人類乳癌細胞T47D細胞中對芳香族碳氫化物受體 (aryl hydrocarbon receptor, AhR) 與雌性激素受體 (estrogen receptor, ERα) 間之訊息交叉 (cross-talk) 之影響進行探討,利用細胞增生及基因表現分析,來比較各種PAHs及TCDD於不同之環境曝露濃度對其雌激素干擾效應之影響。由研究結果顯示,Nap、DMBA、與TCDD於10-13 ~ 10-7 M濃度範圍內單獨存在下,均不會誘發人類乳癌細胞T47D增生,而於同時處理雌性激素E2作用下,與AhR低鍵結力之Nap皆無雌性激素干擾效應,而DMBA與TCDD則於高濃度下具有抑制雌性激素效應,透過與AhR鍵結進而產生抑制雌性激素效應,且DMBA雌性激素干擾效應,均可由同時添加AhR agonist/antagonist, α-naphthoflavone (α-NF) 及CYP1A1抑制劑resveratrol而被抑制,此一結果顯示,DMBA之雌性激素干擾效應會因AhR所調控。由半定量RT-PCR結果顯示,細胞同時曝露於低濃度之DMBA及E2情況下,並不會改變CYP1A1基因之表現量,而PR基因表現量則有明顯增加之情形,若再同時添加AhR agonist, α-NF (10-5 M) ,其CYP1A1基因表現量明顯地被誘發;相反地,細胞曝露於高濃度DMBA及E2情況下,CYP1A1之基因表現量明顯地增加,但PR基因表現量則有被抑制之情形,若同時添加AhR antagonist, α-NF (10-8 M),其CYP1A1基因表現量明顯地被抑制,且PR基因表現量則有誘發之現象,此一結果顯示,DMBA之高低濃度所造成之雌性激素干擾效應,透過ERE-AhR訊息交叉所造成。此外,DMBA於低濃度與E2同時存在下,ERα基因表現量亦明顯地被抑制,於同時添加AhR agonist, α-NF (10-5 M) 下,其抑制ERα基因表現量則消失;反之DMBA於高濃度與E2同時存在下,ERα基因表現量相較於控制組則無改變,但BRCA1基因則明顯被抑制,若再額外添加AhR antagonist, α-NF (10-8 M) ,其ERα基因表現量將被抑制。綜合以上研究結果可確認,於本研究中所使用的PAHs物質與TCDD,皆於10-13 ~ 10-7 M濃度暴露範圍內,單獨存在下並無促進T47D細胞增生作用。而於與雌性激素同時存在下,與AhR親和力較大之物質,能於較高暴露濃度下有抑制雌性激素之作用。本研究亦確認DMBA於高濃度下所造成之抗雌性激素作用,應為透過ERE-AhR訊息交叉而導致增加雌性激素之代謝,以及透過BRCA1基因表現量改變所導致。zh_TW
dc.description.abstractThe primary purpose of this research is to examine the potential of polycyclic aromatic hydrocarbons (PAHs), including naphthalene (Nap), 7,12-dimethylbenz[a]anthracene (DMBA), and the halogenated aromatic hydrocarbons, 2,3,7,8-tertachlorodibenzo-p-dioxin (TCDD), to mediate the estrogen receptor-dependent induction of abnormal cell proliferation in human T-47D breast cancer cells and to investigate the effects of aryl hydrocarbon receptor (AhR) and estrogen receptor (ERα) cross-talk on the ER-dependent altered gene expression. Results indicated that Nap, DMBA, and TCDD did not induce cell proliferation in T47D cells at concentrations ranging from 10-13 to 10-7 M. Co-treatment of Nap (lower binding affinity with AhR) and E2 did not alter the estrogen-induced cell proliferation in T47D cells. In contrast, co-treatment of DMBA, TCDD, and E2 (higher binding affinity with AhR) exhibited antiestrogenic effects in T47D cells. The (anti)estrogenic effects of these compounds correlate with their respective AhR binding affinity. In addition, the antiestrogenic effects of DMBA in T47D cells were completely blocked with the addition of an AhR antagonist, α-naphthoflavone (α-NF) and a CYP1A1 inhibitor, resveratrol. Results from the semi-quantitative RT-PCR analyses confirmed that exposure to DMBA (10-12 M) plus E2 (10-8 M) did not change the expression of CYP1A1 whereas increases in the expression of progesterone receptor (PR) gene were detected in cells. No significant induction of the expression of CYP1A1 gene was detected in cells exposed to DMBA (10-12 M) plus E2 (10-8 M). In contrast, DMBA (10-7 M) plus E2 (10-8 M) induced increases in the expression of CYP1A1 gene and inhibited the expression of PR gene whereas inclusion of α-NF (10-8 M) in the medium abolished these effects. Additionally, co-treatment of DMBA (10-7 M) and E2 did not alter the expression of ERα gene whereas depression of BRCA-1 gene expression was detected. In conclusions, the anti-estrogenic effect of the model compounds selected in this study correlates with their respective AhR binding affinity. The mechanisms by which DMBA exerts its anti-estrogenic effects in human T47D breast cancer cells are likely to mediate via ER-AhR cross-talk and deregulation of BRCA-1 gene expression.en_US
dc.description.tableofcontents摘要 I Abstract III 目錄 VI 圖目錄 X 表目錄 XIII 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 3 2-1-1 PAHS之研究源起與特性 3 2-1-2 PAHs之環境暴露量 4 2-1-3 PAHs與人類癌症之相關性 5 2-2 環境中之污染物 11 2-2-1 naphthalene於環境中流佈與健康危害影響 12 2-2-2 7,12-dimethylbenz[a]anthracene於環境中流佈代謝與健康危害影響 16 2-2-3 四氯戴奧辛於環境中流佈與健康危害影響 19 2-3 AhR、ER之cross-talk與基因之調控 21 2-3-1 芳香烴基受體 (aryl hydrocarbon receptor, AhR) 22 2-3-2 芳香烴基受體之配體 ( AhR ligand) 23 2-3-3 芳香烴基受體之擷抗劑與促進劑 26 2-3-4 雌性激素受體 (estrogen receptor, ER) 29 2-3-5 雌性激素受體之擷抗劑 (ER antagonist) 31 2-3-6 其他基因之調控 32 2-4 PAHs與荷爾蒙干擾作用 39 2-4-1環境荷爾蒙之檢測方式 39 2-4-2 PAHs之雌性激素作用 41 2-4-3 PAHs之抗雌性激素作用 43 2-5 PAHs於細胞週期中之調控機制 49 2-6 研究假設 50 第三章 實驗材料與方法 51 3-1 實驗材料 51 3-1-1 水 51 3-1-2 化學藥品 51 3-1-3 細胞珠來源 52 3-1-4 實驗設備 52 3-2 實驗方法 53 3-2-1細胞培養 53 3-2-2 E-screen分析法 53 3-2-3 Sulforhodamine B assay, SRB 54 3-2-4 反轉錄酶鏈鎖反應分析法(RT-PCR assay) 54 3-2-5 數據分析 57 第四章 實驗結果 58 4-1 E-Screen 分析法之改良 58 4-2 雌性激素干擾效應 59 4-2-1 天然雌激素雌酯二醇誘發細胞增生作用 59 4-2-2 4-hydroxytamoxifen抑制E2誘發細胞增生作用 59 4-2-3 PAHs與HAH物質單獨存在下之細胞增生作用 59 4-2-4 Nap及1-NT與E2共同存在下之雌性激素效應 60 4-2-5 DMBA與E2共同存在下之雌性激素效應 61 4-2-6 TCDD (10-13~10-8 M) 與E2共同存在下之雌性激素效應 61 4-2-7 AhR agonist, α-Naphthoflavone 10-5 M干擾由DMBA導致之雌激素效應 61 4-2-8 AhR antagonist, α-Naphthoflavone 10-8 M干擾由DMBA所導致之抗雌激素效應 62 4-2-9 Reservatrol抑制由DMBA所誘發之細胞增生 63 4-3 PAHs物質對AhR與ER所調控基因之表現 63 4-3-1 DMBA於低濃度(10-12 M)暴露下與同時處理E2 (10-8 M)下之CYP1A1與PR基因表現情況 63 4-3-2 DMBA於高濃度(10-7 M)暴露下與同時處理E2 (10-8 M)下之CYP1A1與PR基因表現情況 64 4-3-3 DMBA於低濃度(10-12 M)暴露下與α-NF (10-5 M)及E2 (10-8 M)同時存在下CYP1A1與PR基因表現情況 64 4-3-4 DMBA於高濃度(10-7 M)暴露下與α-NF (10-8 M)及E2 (10-8 M)同時存在下CYP1A1與PR基因表現情況 65 4-3-5 DMBA於低濃度(10-12 M)暴露下與同時處理E2 (10-8 M)下之ERα基因表現情況 66 4-3-6 DMBA於高濃度(10-7 M)暴露下與同時處理E2 (10-8 M) 下之ERα基因表現情況 66 4-3-7 DMBA於低濃度(10-12 M)暴露下與AhR agonist,α-NF (10-5 M)及E2 (10-8 M)同時存在下ERα基因表現情況 66 4-3-8 DMBA於高濃度(10-7 M)暴露下與α-NF (10-8 M)及E2 (10-8 M)同時存在下ERα基因表現情況 67 4-3-9 DMBA於高濃度(10-7 M)暴露下與同時處理E2 (10-8 M) 下之BRCA1基因表現情況 67 第五章 討論 93 5-1 雌性激素與其受體於人類乳癌細胞之作用 93 5-2 PAHs物質之雌性激素之干擾效應 93 第六章 結論 101 第七章 參考文獻 103zh_TW
dc.language.isoen_USzh_TW
dc.publisher環境工程學系zh_TW
dc.subjectpolyaromatic hydrocarbonsen_US
dc.subject多環芳香族碳氫化合物zh_TW
dc.subjectestrogenen_US
dc.subjectaryl hydrocarbon receptoren_US
dc.subjectestrogen receptoren_US
dc.subject雌性激素zh_TW
dc.subject芳香族碳氫化物受體zh_TW
dc.subject人類乳癌細胞T47Dzh_TW
dc.subject雌性激素受體zh_TW
dc.title以人類乳癌細胞研究多環與鹵化芳香族碳氫化合物之雌性激素干擾效應zh_TW
dc.titleInvestigation of the (anti)estrogenicity induced by polycyclic and halogenated aromatic hydrocarbons in human breast carcinoma cellsen_US
dc.typeThesis and Dissertationzh_TW
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