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研究生:張鳳書
研究生(外文):Chang Feng Shu
論文名稱:Disabled-2在plakoglobin調控的Wnt訊號傳遞路徑中所扮演的角色
論文名稱(外文):The role of Disabled-2 in plakoglobin-mediated Wnt signaling pathway
指導教授:曾慶平
指導教授(外文):Tseng Ching Ping
學位類別:碩士
校院名稱:長庚大學
系所名稱:醫學生物技術研究所
學門:醫藥衛生學門
學類:醫學技術及檢驗學類
論文種類:學術論文
論文出版年:2005
畢業學年度:93
語文別:英文
論文頁數:51
中文關鍵詞:白血病多潛能性惡性造血性白血病細胞株訊號傳遞路徑
外文關鍵詞:Disabled-2Wnt signaling pathwayplakoglobinleukemia
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Wnt訊號傳遞路徑在調控生物的發育及血球前驅細胞的自我更新中扮演一個重要的角色。Armadillo家族成員中的plakoglobin,不僅可以調節細胞間的貼附,並且也可以參予並活化Wnt訊號傳遞路徑。最近的報導指出,plakoglobin與白血病的形成有關。我們初步發現,在血球分化過程中會高度表現的訊號傳遞分子:Disabled-2 (Dab2),可調控由plakoglobin所活化的Wnt訊號傳遞路徑。在此研究中,我們利用人類多潛能性惡性造血性白血病K562細胞株,探討Dab2調控由plakoglobin活化Wnt訊號傳遞路徑的分子機制。我們發現Dab2可藉由N端PTB domain和中間段與plakoglobin結合。由免疫螢光染色發現,當大量表現缺乏中間段DPF motif及C端proline rich domain的Dab2 (△B),或是模擬磷酸化的Dab2 (p82-S24D、p82-S24E),原本在膜上的plakoglobin會轉移到細胞質內。但是若大量表現不能磷酸化的Dab2 (p82-S24A)時,plakoglobin在細胞內的位置就不會受到改變。當大量表現缺乏PTB domain的Dab2:△N時,發現原本在膜上的plakoglobin會跑入細胞質中,甚至部份會進入細胞核內。這些發現指出, Dab2的Serine 24的磷酸化及DPF motif可能參予調控plakoglobin在細胞中的位置,另外,Dab2的PTB domain可以把跑到細胞質內的plakoglobin維持在細胞質中。綜合上述研究結果我們可以了解Dab2是如何調控由plakoglobin所活化的Wnt訊號傳遞路徑,並且更進一步了解Dab2在白血病發生的過程中可能扮演的角色。
Wnt signaling pathway controls many development processes and mediates self-renewal of hematopoietic progenitors. Recent study revealed that plakoglobin, a member of the Armadillo family with dual-function in cell adhesion and signaling, is an activator of Wnt signaling pathway and is involved in leukemogenesis. We further demonstrate that plakoglobin-mediated T cell factor transcriptional activity is modulated by Disabled-2 (Dab2), a signaling molecule that is up-regulated during hematopoietic differentiation. In this study, the molecular mechanism for Dab2 in the regulation of plakoglobin-mediated-Wnt signaling was investigated using human chronic myeloid leukemic cell line K562. We found that Dab2 interacts with plakoglobin through its N-terminal phosphotyrosine binding domain (PTB) and middle region. Immunofluorescent staining revealed that ectopic expression of Dab2, the deletion mutant △B lacking the DPF motif and proline rich domain (PRD), and the phosphorylation mimetic mutants p82-S24D and p82-S24E resulted in plakoglobin translocation from membrane to cytosol. However, the Ser 24 phosphorylation mutant p82-S24A lost its effect on plakoglobin translocation. A Dab2 mutant △N which has a PTB domain deletion resulted in a decrease in membrane-associated plakoglobin concomitantly with an increase of plakoglobin in both cytosolic and nuclear compartments. These findings indicate that serine 24 phosphorylation and DPF/PRD motifs are involved in Dab2-mediated cytosolic translocation of plakoglobin. In addition, the Dab2 PTB motif may be essential for retaining plakoglobin in the cytosol. Together, this study defines the molecular mechanism of Dab2 in plakoglobin-mediated Wnt signaling pathway and contributes to our understanding of Dab2 in the control of leukemogenesis.
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指導教授推薦書

口試委員會審定書

授權書 iii

Acknowledgements iv
Abstract v
Abbreviation vi
Introduction 1
Materials and Methods 13
Results 19
Discussion 24
References 29
Figures 38
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