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研究生:林翠娟
研究生(外文):Cuei-Jyuan
論文名稱:XRCC1之基因多形性與台灣肺癌發生與預後之相關性
論文名稱(外文):The association of XRCC1 genetic polymorphisms and its prognostic value of lung cancer in Taiwan
指導教授:李輝李輝引用關係鄭雅文鄭雅文引用關係
學位類別:碩士
校院名稱:中山醫學大學
系所名稱:醫學分子毒理學研究所
學門:醫藥衛生學門
學類:其他醫藥衛生學類
論文種類:學術論文
論文出版年:2007
畢業學年度:95
語文別:中文
論文頁數:90
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XRCC1是一種鹼基刪除DNA修補(base excision repair)基因。已知該基因有十七個基因多形性,其中位於促進子之T-77C、表現序列6 (Exon 6)之Arg194Trp及表現序列10 (Exon 10)之Arg399Gln等三個位置之基因多形性與人類癌症發生之相關性較常被研究。XRCC1之T-77C基因型中具有T/C 或C/C基因型者較T/T者有較低轉錄活性,會增加罹患肺癌之風險。XRCC1之Arg194Trp與Arg399Gln基因型主要影響與其他蛋白間之結合能力,進而影響XRCC1之DNA修補活性。已知Arg194Trp具有Arg/Trp + Trp/Trp者罹患肺癌之風險較Arg/Arg者為低,但是Arg399Gln基因多形性與罹患肺癌之風險,至今並沒有一致的結論。但已有研究指出具有Gln/Gln基因型者之p53發生突變之危險性較Arg/Arg基因型者為高。因此本研究擬以病例-對照研究探討XRCC1上述三種基因多形性是否與台灣肺癌之發生有關?是否是透過較易引起p53發生突變而形成肺癌?另外,本研究又在病例組探討這三種XRCC1基因多形性是否影響化療藥物的敏感性而影響肺癌患者之預後?
本研究共收集296位肺癌病患之腫瘤周邊正常肺組織與288位非癌症者之血液DNA,以PCR-RFLP方式分析T-77C、Arg194Trp及Arg399Gln之基因多形性,並利用邏輯式回歸分析不同基因型之XRCC1與肺癌發生之相關性。本研究又以Kaplan-Meier方法統計分析不同基因型之XRCC1與肺癌患者預後的相關性。結果僅發現在男性具有TC + CC同時為Arg/Trp + Trp/Trp基因型者,罹患肺癌之風險率是TT同時為Arg/Trp + Trp/Trp基因者的3.51倍(95% CI = 1.028-11.961,p = 0.045),其餘不論何種分層分析皆沒有達到統計上的相關性。在基因多形性與p53突變的相關性分析得到的結果顯示,僅在女性可觀察到具有Gln/Gln基因型者,是Arg/Arg者的4.11倍(95% CI = 1.050-16.117,p = 0.042),而男性僅在TC + CC基因型者p53基因突變之危險性是TT基因型者的1.99倍(95% CI = 0.906-4.353,p = 0.087),但未達到統計上之顯著差異。本研究在分析基因多形性與肺癌患者預後之相關性時發現,具TC + CC基因型的肺癌患者之預後較TT基因型的患者為佳(p = 0.042),因此未來需進一步探討XRCC1之T-77C基因多形性,是否可用來評估肺癌患者之預後。總之,本研究之結果顯示XRCC1之T-77C和Arg194Trp之基因型組合可能與台灣肺癌發生有關。同時分別於女性患者Arg399Gln及男性患者T-77C部分觀察到與p53突變有關,因此推測XRCC1基因多形性可能經由引起p53突變而與肺癌發生有關。


X-ray cross-complementing group 1 (XRCC1) is a base excision repair (BER) gene. There are 17 polymorphisms of XRCC1 been reported. Among these, the association of cancer risk of three polymorphisms of XRCC1 including 5’UTR T-77C, Exon 6 Arg194Trp and Exon 10 Arg399Gln has been extensively investigated. 5’UTR T-77C was association with increased lung cancer risk by reduced transcriptional activity of XRCC1. XRCC1 Arg194Trp and Arg399Gln could interact with some proteins involved in BER to decrease DNA repair activity. It has been shown that subjects with Arg/Trp + Trp/Trp at 194 codon had higher risk of lung cancer than those with Arg/Arg. However, no consistent result was obtained from previous reports regarding Arg399Gln polymorphism. Interestingly, subjects with Gln/Gln at 399 codon exhibited higher p53 mutation frequency than those with Arg/Arg genotype. Therefore, we will conduct a case-control study to understand whether the three genetic polymorphisms of XRCC1 were associated with lung cancer risk? Whether the lung cancer risk caused by these genetic polymorphisms was mediated through the increase of p53 mutation? Additionally, whether patient’s survival could be influenced by these genetic polymorphisms of XRCC1?
To answer those questions, 296 lung cancer cases and 288 non-cancer hospital controls were enrolled to determine the genotypes of XRCC1 by PCR-RFLP. Multiple logistic regression was performed to statistically analyzed understanding which genotype of XRCC1 was associated with lung cancer risk. The association of XRCC1 genotypes with patient’s survival was statistically analyzed by Kaplan-Meier method. Our data showed that male subjects with combination of TC + CC and Arg/Trp + Trp/Trp genotype had 3.51-fold lung cancer risk of those with the combination of TT and Arg/Trp + Trp/Trp genotype (95% CI = 1.028-11.961, p = 0.045). We also observed that female cases with Gln/Gln genotype exhibited a significantly higher p53 mutation frequency than those with Arg/Arg genotype (OR = 4.11, 95% CI = 1.050-16.117, p = 0.042). On the other hand, male cases with T/C + C/C genotype had more common to have p53 mutation than those with T/T genotype (OR = 1.99, 95% CI = 0.906-4.353, p = 0.087), but it did not reach statistically significant. Among the prognostic influence of XRCC1 polymorphisms, we only observed that patients with TC + CC genotype had longer survival time than those with T/T genotype (p = 0.042). Taken together, these results suggest that the combined T-77C and Arg194Trp genotype of XRCC1 could increase the risk of lung cancer in Taiwanese population. The association of XRCC1 genetic polymorphisms with lung cancer risk could partly mediate through the increase of p53 mutation.


壹、中文摘要..............................................1
貳、英文摘要..............................................3
參、文獻綜論..............................................5
一、肺癌之流行病學及形成機制..............................5
1. 肺癌流行病學特徵......................................5
2. 肺癌的組織學型態......................................5
3. 肺癌之危險因子........................................6
4. 性別差異與肺癌發生之相關性...........................11
二、反應性氧(reactive oxygen species,ROS)...............12
1. ROS簡介..............................................12
2. ROS與腫瘤形成之相關性................................14
三、鹼基刪除修復(Base excision repair,BER)..............15
四、XRCC1 (X-ray repair cross-complementing group 1).....16
1. 基本的分子結構與蛋白特性.............................16
2. XRCC1基因多形性與癌症相關性..........................18
五、p53..................................................20
1. 基本的分子結構與蛋白特性.............................20
2. p53在肺癌扮演之角色..................................21
六、基因多形性與預後之相關研究...........................23
肆、研究動機.............................................25
伍、實驗材料.............................................26
一、檢體收集.............................................26
二、材料與藥品...........................................26
陸、實驗方法.............................................28
一、肺臟組織DNA萃取與純化................................28
二、血液DNA之萃取與純化..................................29
三、聚合酶連鎖反應-限制片段長度多形性(PCR-RFLP)..........30
四、DNA定序分析..........................................33
五、統計分析.............................................35
柒、結果與討論...........................................36
一、肺癌個案與非癌症對照組之年齡、性別、抽菸配對分佈.....36
二、XRCC1個別基因型與肺癌發生之危險性評估................36
三、XRCC1基因型組合與肺癌發生之危險性評估................42
四、XRCC1基因型與肺癌p53發生突變之危險性評估.............46
五、XRCC1個別基因型與肺癌預後之相關性分析................50
捌、參考文獻.............................................53
玖、表與圖...............................................63
壹拾、附錄...............................................89

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