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研究生:朱書呈
研究生(外文):Shu-Cheng Chu
論文名稱:探討戴奧辛暴露與兒茶酚-O-甲基轉移酶 (COMT) 基因多型性與雌性激素醌類代謝物之蛋白質胼合物之相關性
論文名稱(外文):Investigation of the Associations of the Background Levels of Estrogen Quinone-Derived Protein Adducts with Dioxin Exposure and Catechol-O-Methyl Transferase (COMT) Genetic Polymorphisms
指導教授:林伯雄林伯雄引用關係
指導教授(外文):Po-Hsiung Lin
口試委員:李立安林嬪嬪
口試委員(外文):Lih-Ann LiPin-Pin Lin
口試日期:2015-07-10
學位類別:碩士
校院名稱:國立中興大學
系所名稱:環境工程學系所
學門:工程學門
學類:環境工程學類
論文種類:學術論文
論文出版年:2015
畢業學年度:103
語文別:中文
論文頁數:116
中文關鍵詞:戴奧辛基因多型性雌性激素
外文關鍵詞:dioxinGenetic Polymorphismestrogen
相關次數:
  • 被引用被引用:0
  • 點閱點閱:184
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  • 下載下載:10
  • 收藏至我的研究室書目清單書目收藏:1
本研究第一部分之主要目的為運用經穩定基因轉殖之人類肺腺癌細胞株CL1-5 (TO-AhR) 、CL1-5 (TO- AhR/ERα) 、CL1-5 (TO-ERα) 探討於四氯戴奧辛 (2,3,7,8-tetrachlorodibenzo-p-dioxin,TCDD) 之暴露下,對雌性激素之醌類活性代謝物 (E2-2,3-Q、E2-3,4-Q) ,所形成之蛋白質胼合物背景值之相關性。實驗結果顯示,共同添加TCDD (10 nM) 及E2之實驗中,各組細胞樣本 CL1-5 (TO-AhR)、CL1-5 (TO-AhR/ERα)、CL1-5 (TO-ERα) ,在雌性激素濃度為1 nM、反應時間為 6 h 條件下,CL1-5 (TO-AhR/ERα) 及CL1-5 (TO-ERα) 之雌性激素醌類細胞質蛋白質(cytoplasmic protei, Cp) 胼合物E2-3,4-Q-2-S-Cp背景值相對於CL1-5 (TO-AhR) 約高出1.5 倍及1.4 倍 (**p<0.01) 。 各細胞樣本 CL1-5 (TO-AhR) 、CL1-5 (TO-AhR/ERα)、CL1-5 (TO-ERα) 之數據,所形成之蛋白質胼合物與添加之E2濃度,均無劑量效應之相關性。
本研究第二部分研究則是探討台灣女性乳癌病人其雌性激素代謝基因兒茶酚-O-甲基轉移酶 (catechol-o-methyl transferase, COMT) 之基因多型性與蛋白質胼合物背景值之相關性,進而作為台灣女性罹患乳癌之風險預測指標。單一核苷酸基因多型性 (single nucleotide polymorphisms, SNPs) 實驗結果顯示,彰化基督教醫院乳癌病人 (n=146) 之雌性激素代謝基因 (COMT Val158Met) 之對偶基因 (variant alleles) 頻率為27.7% 。 經由勝算比 (Odd ratio)之計算,乳癌病人與健康婦女族群間並無統計上之差異顯著,亦即COMT Val158Met基因型之變異與台灣女性罹患乳癌之風險無相關性。進一步研究發現,此基因之變異型與人體內雌性激素醌類蛋白質胼合物背景值具相關性,其中原生型 (Val/Val)之E2-3,4-Q-2-S-Hb及E2-2,3-Q-4-S-Hb背景值分別高於攜帶變異型 (Val/Met+Met/Met) 1.2倍 (p<0.01) 及1.2倍 (p<0.05) 。
The objective of the first part of this study was to investigate the effect of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) exposure on the formation of estrogen quinone-derived cytoplasmic protein (Cp) adducts, including estrogen-2,3-quinones (E2-2,3-Q) and estrogen-3,4-quinones (E2-3,4-Q) , in human lung adenocarcinoma cell line CL1-5 (TO), including CL1-5 (TO-AhR) , CL1-5 (TO- AhR/ERα), and CL1-5 (TO-ERα). Results indicated that the background level of E2-3,4-Q-2-S-Cp was significantly increased in CL1-5 (TO-AhR/ERα) and CL1-5 (TO-ERα) cells treated with TCDD (10 nM) plus E2 (1 nM) when compared to CL1-5 (TO-AhR) cells (~1.5-fold, **p<0.01) and (~1.4-fold, **p<0.01), respectively. All of the sample (CL1-5) did not display time- and concentration-dependent production of estrogen quinone-derived protein adduct. .
The second part of this study was to investigate the associations of the genetic polymorphisms of catechol-o-methyl transferase (COMT) with the background level of estrogen quinone-derived protein adducts and the risk of developing breast cancer in Taiwanese women. Results from analysis of single nucleotide polymorphisms indicated that the frequency of variant allels of estrogen metaboism gene COMT Val158Met were estimated to be 27.7% for breast cancer patients (n=146). Similar observation was also detected in the corresponding healthy controls. This finding suggest that variant allels of COMT Val158Met do not associate with the risk of developing breast cancer in Taiwanese women. However, we did observe the levels of estrogen quinone-derived adducts of E2-3,4-Q-2-S-Hb and E2-2,3-Q-4-S-Hb in breast cancer patients with wild-type COMT (Val/Val) were about 1.2-fold (p<0.05) greater than those with variant alleles (Val/Met+Met/Met).
摘要 I
Abstract II
縮寫表 III
縮寫表 IV
目次 V
表目次 VII
圖目次 X
第一章 前言 1
1.1. 研究緣起 1
1.2. 研究目的 2
第二章 文獻回顧 3
2.1. 戴奧辛 (Dioxins) 3
2.2. AhR與ER之交互作用與基因之調控 19
2.3. 雌性激素 (Estrogen) 25
2.4. 蛋白質胼合物 (protein adducts) 33
第三章 實驗架構與研究假設 37
3.1. 實驗設計架構 37
3.2. 實驗假設 41
第四章 實驗材料與方法 44
4.1. 實驗材料 44
4.2. 實驗方法 47
4.3. 氣相層析質譜儀分析條件 51
4.4. 聚合酵素鏈鎖反應 (Polymerase Chain Reaction, PCR) 產物之定序確認 52
4.5. 統計分析 54
第五章 實驗結果 55
5.1. 分析人類肺腺癌細胞株CL1-5 (TO) 共同添加TCDD及E2後所生成之雌性激素醌類蛋白質胼合物之背景值 (n=70) 59
5.2. 分析人類肺腺癌細胞株CL1-5 (TO) 未添加E2後所生成之雌性激素醌類蛋白質胼合物背景值 (n=3) 62
5.3. 分析人類肺腺癌細胞株CL1-5 (TO) 預處理TCDD並單獨添加E2後所生成之雌性激素醌類蛋白質胼合物背景值 (n=4) 63
5.4. 分析彰化基督教醫院提供乳癌病人之單一核甘酸多型性 (SNPs) –COMT Val158Met之背景值 64
第六章 結果與討論 66
6.1. 人類肺腺癌細胞株CL1-5 (TO) 共同添加TCDD及E2後所生成之雌性激素醌類蛋白質胼合物之背景值 66
6.2. COMT Val158Met之基因多型性 72
6.3. 彰化基督教醫院乳癌病人體內雌性激素 (E2) 蛋白質胼合物背景值與SNPs之相關性 75
6.4. 單一核苷酸基因多型性 (SNPs – COMT Val158Met、CYP1A1 T3801C、CYP1B1 Leu432Val、NQO1 C609T) 與蛋白質胼合物之相關性分析 90
第七章 結論與建議 96
參考文獻 99
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11. 王讚源〈思考的盲點-思想方法探討之一〉,《中華文化復興月刊》,民國七十六年八月,二十二卷八期。
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13. 李震〈王充與無神主義〉,《哲學與文化》,民國七十八年五月,第一八○期,頁四十至五一。
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