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    Please use this identifier to cite or link to this item: http://ir.lib.ncu.edu.tw/handle/987654321/6318

    Title: Galectin-1蛋白對細胞生長於幾丁聚醣膜上的影響及調控機制之研究;The study of mechanism of galectin-1 regulates cells proliferation on chitosan membrane
    Authors: 張鈺媖;Yu-Ying Chang
    Contributors: 生命科學研究所
    Keywords: 幾丁聚醣膜;Galectin-1;Galectin-1;Chitosan membrane
    Date: 2003-07-04
    Issue Date: 2009-09-22 10:17:30 (UTC+8)
    Publisher: 國立中央大學圖書館
    Abstract: 幾丁聚醣是由螃蟹、龍蝦、蝦子這類甲殼類動物外殼中的幾丁質所衍生而來的。在臨床醫學上,幾丁聚醣膜已被使用當作具有半滲透性的生物敷料。雖然幾丁聚醣膜是不具有細胞毒性的,但有些細胞種類 (如: 3T3纖維母細胞)就是無法於幾丁聚醣膜表面進行貼附及增生的作用。近年有許多的研究已指出galectin-1 (Gal-1, 是一種?-半乳糖苷結合的蛋白)對於加速細胞貼附於細胞外基質及誘導細胞的增生而言,扮演重要的角色。因此,在本篇的研究中,我們將探討Gal-1蛋白影響3T3細胞在幾丁聚醣膜上生長的情形。發現當3T3細胞中的Gal-1蛋白被大量表現之下,無法促進細胞生長於幾丁聚醣膜上。然而,將Gal-1蛋白吸附在幾丁聚醣膜上時,則能明顯的加強細胞在幾丁聚醣膜上增生的效率。若處理Gal-1單株抗體及Thiodigalactoside (TDG, lectin的抑制劑),則能抑制Gal-1對細胞增生的影響,此說明了確實是幾丁聚醣膜上所吸附的Gal-1蛋白對細胞的增生所產生的特殊影響。另外,利用免疫標定的方式也偵測到當3T3或CHOA細胞培養在Gal-1吸附的幾丁聚醣膜上時,會明顯的降低細胞內某些蛋白質酪胺酸磷酸化的表現;當使用NaF (絲胺酸/息寧胺酸去磷酸酶抑制劑)處理細胞,NaF明顯的抑制3T3細胞在Gal-1蛋白吸附的幾丁聚醣膜上的增生。此結果表示由Gal-1所誘導的細胞貼附及增生,主要是藉由改變蛋白質磷酸化所產生的訊號傳遞,以進而促進細胞在幾丁聚醣膜上的生長。 Chitosan is a derivative of chitin extracted from the exoskeleton of lobsters, crabs and shrimps. In clinical medicine, chitosan membrane (a tissue-engineered chitin membrane) has been used as a semi-permeable biological dressing. Although the chitosan membranes showed no cytotoxicity, some cell types (e.g. 3T3 cell) were unable to attach and proliferate on the surface of chitosan membrane. A number of studies suggested that galectin-1 (Gal-1, a b-galactoside-binding protein) plays an important role in enhancing the adhesion of cells to extracellular matrix and inducing cell proliferation. Therefore, we here explored the applicability of Gal-1 on enhancing the growth of 3T3 cell on the surface of chitosan membrane. The results showed that over-expression of Gal-1 could not enhance 3T3 cell proliferation on chitosan membrane. However, coating the chitosan membrane with recombinant Gal-1 proteins significantly enhanced the efficiency of 3T3 cell proliferation. However, this effect was inhibited by Gal-1 monoclonal antibody and thiodigalactoside (TDG, a lectin inhibitor), suggesting the specific effect of Gal-1 on the proliferation of 3T3 cell. Moreover, immunoblotting detected a significant reduction in tyrosine phosphorylation of several proteins in 3T3 fibroblast and CHOA cell growing on Gal-1-coated chitosan membrane. The results also showed that NaF (a serine/threonine phosphatase inhibitor) dose-dependently inhibits 3T3 cell proliferation on Gal-1-coated chitosan membrane. These findings support a proposed role for altered levels of protein phosphorylation in Gal-1-mediated cell attachment and proliferation.
    Appears in Collections:[生命科學研究所 ] 博碩士論文

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