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研究生:白竣翔
研究生(外文):Chun-Hsiang Pai
論文名稱:探討兒童腦部生殖細胞腫瘤的MGMT啟動子甲基化狀況及蛋白質表現程度
論文名稱(外文):Promoter methylation status and expression of O6-Methylguanine-DNA-Methyltransferase (MGMT) in pediatric brain germ cell tumors
指導教授:陳燕彰陳燕彰引用關係
指導教授(外文):Yann-Jang Chen
學位類別:碩士
校院名稱:國立陽明大學
系所名稱:生命科學暨基因體科學研究所
學門:生命科學學門
學類:生物學類
論文種類:學術論文
論文出版年:2009
畢業學年度:97
語文別:英文
論文頁數:38
中文關鍵詞:腦部生殖細胞腫瘤啟動子甲基化
外文關鍵詞:brain germ cell tumorMGMTPromoter methylation
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一些烷基化藥物如temozolomide被利用在治療癌症,尤其在治療腦瘤有很大的功用。但這些烷基化藥物對癌細胞的毒殺效用卻會因細胞中O6-methylguanine-DNA methyltransferase (MGMT)的修補作用而被減弱。MGMT是細胞中的DNA修補酵素, 它能移除經由烷基化藥劑作用而在鳥嘌呤的O6部位形成具致突變性的烷基加合物。先前研究顯示, MGMT基因啟動子部位的CpG islands如發生異常的甲基化現象,將使得此基因無法正常行使轉錄功能而造成MGMT蛋白質表現量降低。而在許多癌症細胞中都有發現到此一情形。所以能夠了解癌症病患其癌細胞中MGMT蛋白質的表現及其基因啟動子甲基化狀況,對於患者診療及用藥都有很重要的助益。腦部生殖細胞腫瘤形成原因是因為生殖細胞在細胞發育階段時不正常的遷移,造成該細胞坐落在腦部而並非在正確的睪丸和卵巢等部位。此疾病大約有一半的比率發生在十到二十歲的兒童和青少年之間,而目前對此疾病的治療是以放射性治療和化學療法為主。經由台北榮總所提供的21個兒童腦部生殖細胞腫瘤標本,我們進行亞硫酸氫鹽甲基化專一性聚合酵素鏈鎖反應和亞硫酸氫鹽定序法以了解其MGMT 基因啟動子的CpG islands 甲基化狀況,並由免疫組織化學染色法的結果去觀察其MGMT蛋白質的表現情形, 希望從中了解此疾病是否適合使用烷基化藥物治療。經由研究顯示,在聚合酵素鏈鎖反應中,二十一個患者有五位(25%)在啟動子部位出現甲基化反應。從亞硫酸氫鹽定序法中觀察到,在104個CpGs中,含有第1到第26個CpGs的區域有較高比率的甲基化現象,最低為12%,最高則達到87%,其餘部分大概都在10%以下。在免疫組織化學染色法的結果發現僅有一位完全失去MGMT蛋白質表現,其餘患者則有高低不等的蛋白質表現。而在這些患者之中, MGMT 基因啟動子的甲基化狀況和蛋白質表現並沒有存在顯著的相關性。因此,除了啟動子甲基化,或許還有其他機制調控MGMT蛋白質的表現,¬而這些都需要進一步的舉證。
Alkylating drugs such as temozolomide is used in the treatment of several human cancers, but the cytotoxicity of these anticancer drugs results from the alkylation of DNA is strongly attenuated by the repair ability of O6-methylguanine-DNA methyltransferase (MGMT). The cellular DNA repair protein MGMT, a DNA repair enzyme that removes the mutagenic alkyl-adducts from the O6-position of guanine and thereby causes resistance to alkylating agents. Aberrant methylation of CpG islands located in the promoter region of MGMT gene is associated with transcriptional inactivation of this DNA repair gene and result in low expression of the DNA repair enzyme. In some cancer cells, the expression of MGMT is silenced due to abnormal promoter methylation. Therefore, to realize the MGMT expression and its promoter methylation status is important for advancing benefit of these chemotherapy drugs in cancer patients. Brain germ cell tumor arises from the abnormal migration of germ cells which should be located on testis or ovarian in embryo development stage. Around half of these tumors occur in young people between 10 and 20 years old. The major treatments for brain germ cell tumors are radiation and Chemotherapy. In order to realize whether it is suitable to treat brain germ cell tumor patients with alkylating drugs, we performed immunohistochemistry (IHC), bisulfite methylation specific PCR (MSP) and bisulfite sequence on 21 pediatric brain germ cell tumors from Taipei Veterans General Hospital to investigate the relation between the expression of MGMT and the CpG methylation within their promoters in brain germ cell tumors. In our study, hypermethylation of MGMT was detected in five out of twenty-one (25%) cases in MSP. Sequence-1(CpG1~26) has the highest methylation frequency from 12 to 87% in each case, and the other regions are almost under 10%. In IHC, just one case lost the expression of MGMT, and the others had the different-grade expression of MGMT. The relation between MGMT expression and promoter methylation is not concordant. In addition to promoter methylation, some mechanisms may regulate the expression of MGMT, and this needs further evaluation.
中文摘要………………………………………………2
Abstract....................................4
Introduction…………………………………………6
Study aims………………………………………….11
Materials and methods……………………………12
Results………………………………………………16
Discussion………………………………………….19
Reference……………………………………………23
Tables……………………………………………….28
Figures………………………………………………35
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