研究生: |
陸一麟 LU, YI-LIN |
---|---|
論文名稱: |
GAS7基因於肺癌之分子變異及臨床相關性研究 Molecular Alteration Analysis of GAS7 Gene and Its Clinical Significance in Lung Cancer |
指導教授: |
王憶卿
Wang, Yi-Ching 李桂楨 Lee, Guey-Jen |
學位類別: |
碩士 Master |
系所名稱: |
生命科學系 Department of Life Science |
論文出版年: | 2008 |
畢業學年度: | 96 |
語文別: | 中文 |
論文頁數: | 69 |
中文關鍵詞: | 肺癌 、抑癌基因 、生長抑制基因 |
英文關鍵詞: | lung cancer, tumor supressor gene, GAS7 gene |
論文種類: | 學術論文 |
相關次數: | 點閱:146 下載:1 |
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自1982年起,癌症即為台灣地區十大死亡原因之第一位,其中肺癌不論在女性或男性都高居癌症死亡率的首位,儘管目前醫學已相當進步,但對於分子致癌機制仍未完全釐清。目前所知,癌症形成的原因,是由於多重基因發生變異所造成,其中大家所熟知和癌症有關的為抑癌基因 (tumor suppressor gene) 及致癌基因 (oncogene)。因此,抑癌基因變異的研究有助於了解癌症形成的機制。 研究目的:Growth arrest-specific 7 (GAS7) 這個蛋白質為GAS這個基因家族的其中一個成員,主要功能可能與調控細胞週期有關;前人研究報導顯示,GAS7可能在細胞中扮演一抑癌基因的角色。因此,本研究目的在探討GAS7基因在台灣地區非小細胞肺癌 (non-small cell lung cancer) 病人細胞中之變異情形:利用免疫組織化學染色法,觀察病人組織切片中GAS7蛋白表現情形,再以反轉錄—聚合酵素鏈反應分析組織細胞中mRNA轉錄是否異常,續以甲基化為基礎的定序反應以及微衛星基因座鑑定法分別偵測GAS7基因的促進子過度甲基化頻率和基因座缺失頻率。 結果: 本研究以免疫組織化學染色法發現75位NSCLC病人中GAS7蛋白低表達頻率為57.3% (43/75),以31位病人之組織進行西方轉漬法發現三種主要的蛋白形式,分別依分子量大至小為GAS7C、7B和7A,其GAS7C蛋白低表現之比例為48.4% (15/31),GAS7B蛋白低表現之比例為40.7% (11/27),而GAS7A蛋白幾乎不表現;GAS7 mRNA isoform: GAS7C和GAS7B低表達頻率分別為20.9% (19/91)和32.6% (30/92),而GAS7C和7B啟動子甲基化頻率分別為16.7% (6/36) 和65.6% (21/32)。GAS7基因座缺失的頻率在位於基因上、下游兩微衛星序列取聯集後為20.7% (18/87)。GAS7蛋白質/mRNA、GAS7C和7B之mRNA/啟動子甲基化的數據彼此間都呈現統計上的顯著相關性 (P<0.05)。利用西方轉漬膠片中相對於GAS7C分子量之蛋白進行膠體內水解、trypsin反應及質譜分析,本研究證實其為GAS7C蛋白。同時,我們藉由不同肺細胞株一系列的基因及蛋白質實驗,我們發現GAS7蛋白不同isoforms如GAS7C和7B,表達的量與細胞內的位置也都不同。 結論:本研究證實GAS7基因在台灣地區肺癌形成過程中扮演一個類似抑癌基因的角色,其中GAS7C為主要的變異蛋白,其變異主要機制為啟動子過度甲基化及基因座缺失,而 GAS7C 蛋白則可以在生長較為緩慢之正常肺細胞表現,此研究也是首篇證實GAS7C isoform可以在人類細胞中表現之論文。
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