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Title: 半乳糖凝集素誘導醣化作用在肺癌惡性轉移之功能性蛋白質體學研究
Authors: 陳泰源;楊崑德;陳水田;余忠仁
Contributors: 國立臺灣海洋大學:食品科學系
Date: 2009-08
Issue Date: 2014-10-02T08:30:32Z
Publisher: 行政院國家科學委員會
Abstract: 摘要:從臨床統計資料發現,第四型岩藻糖基轉移酶大量表現的肺癌病人的五年存活率比一般的肺癌病人更差,探究其原因可能為大量肺癌細胞遠端轉移至其他器官導致。因此,本實驗設計先將第四型岩藻糖基轉移酶過量表現至肺癌細胞(A549F),再將對照組(A549M)與實驗組(A549F)進行癌細胞惡性轉移外顯因子的特性分析。以差異性蛋白質體技術分析第四型岩藻糖基轉移酶大量表現於肺癌細胞後之蛋白質的綜觀表現。結果發現半乳糖凝集素-1 (galectin-1, Gal-1),熱休克蛋白質B1 (heat-shock protein 27 kD, HSPB1)和 peroxiredoxin-1 (PRDX-1)表現量明顯增加。進一步以半乳糖凝集素-1的RNA干擾片段對半乳糖凝集素-1進行基因默化。目前已完成最佳實驗條件,接下來後續將將探討半乳糖凝集素-1低量表現後對肺癌細胞惡性轉移外顯因子的特性,以釐清因第四型岩藻糖基轉移酶大量表現的肺癌細胞中半乳糖凝集素-1大量表現的生理意義。
Abstract:The molecular mechanism of metastasis remains largely unknown presently. Fucosylation is highly involved in invasion, metastasis, and poor prognosis in several types of cancers. In the present study, A549 lung cancer cells with highly (A549F) and poorly (A549M) metastatic ability were used to identify lung cancer metastasis-related proteins. The A549F was created by overexpression of α-1,3-fucosyltransferase IV (FUT4) in A549 cells. The potent migration and adhesion power verified metastatic phenotypes in A549F cells. The detergent-based extraction achieved membrane protein lineage. The 2D fluorescent differential gel electrophoresis (DIGE) approach was selected to overview the global membrane proteome change. The significantly up-regulated proteins in highly metastatic A549F cell were related to the angiogenesis and metastasis-associated galectin-1 (Gal-1), the molecular chaperon, heat-shock protein 27 kD (HSPB1), and the stress-related protein, peroxiredoxin-1 (PRDX-1). All proteins were validated by western blots and indicated a good correlation to DIGE. The decreased of invasion ability in FUT4-transfectant A549 cells were exhibited through galectin-1 knock-down experiment. This study can therefore assist in defining the membrane proteome changes that occur in FUT4 overexpressed A549 cells, and implying galectin-1 as possible target for treating lung cancer metastasis.
Relation: NSC98-2311-B019-004-MY2
Appears in Collections:[Department of Food Science] Research Reports

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