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系統識別號 U0026-2508201121015100
論文名稱(中文) 核苷酸切除修補系統之基因多型性、井水砷暴露與膀胱癌之相關性研究
論文名稱(英文) DNA repair gene polymorphisms in the necleotide excision repair pathway, arsenic exposure in well water, and bladder cancer
校院名稱 成功大學
系所名稱(中) 環境醫學研究所
系所名稱(英) Institute of Environmental and Occupational Health
學年度 99
學期 2
出版年 100
研究生(中文) 潘思錞
研究生(英文) Sih-Chun Pan
學號 s76984105
學位類別 碩士
語文別 中文
論文頁數 72頁
口試委員 指導教授-郭浩然
口試委員-王應然
口試委員-張志欽
口試委員-郭炤裕
中文關鍵字   DNA 修復基因  單一核甘酸基因多型性  膀胱癌  交互作用 
英文關鍵字 arsenic  DNA repair gene  polymorphism  bladder cancer  interaction 
學科別分類
中文摘要 核苷酸切除修復(necleotide excision repair;NER)是人體DNA修補系統中其中一種修復機制,主要是修補因外在環境因素所造成的損傷,以維持基因體的穩定性,在癌症預防上扮演著相當重要的角色。DNA修復基因的多型性可能會影響宿主修復DNA損傷的能力,因而與癌症的易感性有關。過去研究指出,砷會降低NER系統內基因的表現與修復功能,被認為可能是砷導致癌症的一種致癌機制。過去研究發現臺灣西南沿海一帶,尤其在嘉義、臺南地區的地下水含有高濃度的砷,也發現膀胱癌的發生率在含砷濃度最高的烏腳病地區為全臺之冠。因此本研究從NER系統中挑選出三個重要的修復基因(XPC、XPD、ERCC1),探討此三種基因的多型性與砷暴露對於膀胱癌之獨立作用及環境基因的交互作用。本研究採病例對照研究設計,膀胱癌組個案數有104位,對照組個案數有265位。我們以問卷收集基本人口學資料與生活習慣、相關危險因子,另用居住區域地下水中砷濃度及飲用地下水時間做個人砷暴露指標,並收集血液樣本萃取DNA利用聚合酵素連鎖反應、限制酵素片段長度多型性(PCR-RFLP)進行基因型分析。我們利用多變項邏輯斯迴歸分析,發現高年齡、男性、低教育程度、抽菸、及高砷暴露均顯著增加罹患膀胱癌的風險,且砷暴露與膀胱癌具有顯著的劑量效應關係。ERCC1 Asn118Asn(TC)基因多型性與膀胱癌無顯著相關性,但會修飾砷暴露與膀胱癌間的風險;在同樣為高砷暴露下的人,帶基因型(T/C+C/C)的勝算比較帶T/T基因型者高(校正後勝算比與95%信賴區間分別為5.4,2.7-11.0與2.3,1.2-4.6)。而XPD Lys751Gln及XPC Ala499Val基因多型性與膀胱癌有邊緣性的顯著關係,帶有變異型的基因會增加罹患膀胱癌的風險,但非砷暴露與膀胱癌相關性的修飾因子。利用THESIS3.1執行ERCC1 Asn118Asn、XPD Lys751Gln 的單套基因型分析,發現攜帶兩個變異對偶基因組的勝算比3.7 (95%信賴區間0.1-20.1)。評估環境暴露(抽菸、砷暴露)與危險性基因型對膀胱癌風險的共同作用,結果發現環境因子的作用強於基因的作用,並發現隨著危險因子越多,膀胱癌危險性就越高,呈顯著的劑量效應關係。本研究發現在西南沿海飲水含中、高濃度砷地區之居民ERCC1 Asn118Asn 基因型為T/C或C/C者其罹患膀胱癌的風險會比帶T/T基因型的人高;若抽菸又帶有較多的危險基因型者,其罹患膀胱癌的風險是飲水含低濃度砷、非抽菸且帶較少危險基因型者的15.1倍(校正後勝算比為15.1; 95%信賴區間2.5-93.1)。這也表示若減少這些危險因子,可降低膀胱癌的發生機率。本研究確定砷與膀胱癌的相關危險性,更發現若同時有抽菸及帶較多的危險基因型者其罹癌的風險最高,因此從事公共衛生相關人員應針對這些高危險群做戒菸的宣導,並對易感性族群更加注意與提高警覺。
英文摘要 DNA adducts caused by environmental and occupational chemical carcinogens are repaired primarily through the nucleotide excision repair (NER) pathway. Arsenic is well established as a human carcinogen, and previous research indicated that arsenic may act through the inhibition of DNA repair mechanisms. Some polymorphisms in DNA repair genes may result in decreases in DNA repair efficiency and thus be associated with cancer risk. The current study determines the prevalence of polymorphisms of ERCC1 Asn118Asn, XPD Lys751Gln and XPC Ala499Val in the NER pathway and their correlations with the risk of bladder cancer in Taiwan and evaluates whether these polymorphisms interact with arsenic exposure to increase susceptibility to bladder cancer. We recruited 104 bladder cancers cases and 265 cancer-free controls. Detailed data on demographic, life style, and environmental factors were collected using a questionnaire. Genotypes were determined using PCR-RFLP analyses. The data were analyzed using logistic regression models, adjusting for relevant covariates. We found older age, male gender, lower education level, and cigarette smoking were significantly risk factors of bladder cancer. Moreover, arsenic exposure was significantly associated with bladder cancer, with a dose-response relationship (p for trend <0.05). The three polymorphisms were in Hardy-Weinberg disequilibrium. We found no associations between ERCC1 Asn118Asn and bladder cancer were found, but the genetic polymorphisms may modify the associations between arsenic and bladder cancer (adjusted odds ratio [OR]=5.4; 95% confidence interval [CI], 2.7-11.0 comparing to homozygous wild type) in the high arsenic exposure group. We found that the C allele of XPD Lys751Gln(AC) and T allele of Ala499Val (CT) were associated with increased risks of bladder cancer (OR=1.6; 95% CI, 0.9-2.8 and OR=1.2; 95% CI, 0.9-1.7, respectively), but we did not observe a significant gene-environment interaction between these polymorphisms and arsenic exposure. The highest bladder cancer risk of 15.1 was observed in those with all environmental risk factors (cigarette smoking and arsenic exposure) and two or more high risk genotypes of ERCC1, XPD and XPC. Our findings suggest that the T/C+C/C of ERCC1 Asn118Asn is a high risk genotype of bladder cancer among residents of arsenic endemic area in southwestern Taiwan and that cigarette smoking, arsenic and high-risk genotypes of ERCC1, XPD and XPC have a significant joint effect on bladder cancer.

論文目次 目 錄
壹、前言..1
第一節 研究背景..1
第二節 研究目的..2
第三節 研究架構..2
第四節 本研究重要性..2
貳、文獻探討..4
第一節 砷的特性..4
第二節 砷暴露與代謝途徑..4
第三節 砷引起的健康效應..5
一、砷對身體系統之非致癌性影響..5
二、砷之致癌性..7
第四節 膀胱癌流行病學研究..8
第五節 無機砷抑制DNA修復能力..10
第六節 DNA 修補系統酵素..11
一、ERCC1(excision repair cross-complementing group 1)..11
二、ERCC2/XPD(xeroderma pigmentosum complementation group D) ..13
三、XPC(xeroderma pigmentosum complementation group C)..14
叁、材料與方法..16
第一節 研究設計與架構..16
第二節 研究對象的來源與選取條件..17
一、病例組..17
二、對照組..17
第三節 資料收集方法與步驟..17
一、問卷內容..17
二、血液檢體收集..18
第四節 砷暴露指標評估..18
一、依地理區域與飲用水時間、年數劃分砷暴露..19
第五節 實驗方法..20
一、萃取DNA..20
二、聚合酶連鎖反應 (polymerase chain reaction, PCR)..20
三、限制片段長度多型性分析(restriction fragment length polymorphism, RFLP)..21
四、ERCC1 Asn118Asn 基因型判定..22
五、XPD Lys751Gln基因型判定..23
六、XPC Ala499Val基因型判定..24
七、定序分析..25
八、降低訊息誤差之方法..26
第六節 統計分析..26
肆、研究結果..27
第一節 基本人口學變項..27
第二節 危險因子與膀胱癌關係..28
第三節 基因多形性與膀胱癌間相關性..28
第四節 砷暴露與基因多形性之交互作用..30
第五節 單套基因型(haplotype)與膀胱癌相關性..31
第六節 環境暴露與危險性基因型對膀胱癌風險的聯合效應..32
伍、討論..34
第一節 病例組與對照組描述..34
第二節 膀胱癌危險因子..35
第三節 基因型、砷暴露與膀胱癌的風險探討..36
第四節 單套基因型、砷暴露與膀胱癌風險 ..39
第五節 綜合環境因子與危險基因型對膀胱癌之風險評估..39
陸、結論與建議..41
第一節 結論..41
第二節 限制與建議..41
柒、參考文獻..43

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