TGF-β活化激酶1（TAK1）是一個屬於MAPKKK家族的蛋白激酶，研究顯示其參與了JNK、p38及NFκB相關的訊息傳遞路徑。TAK1結合蛋白TAB1可以增進TAK1的激酶活性。此外，TAK1也與發炎、先天性免疫反應及許多類型癌症的惡化有關。近來的研究顯示趨化激素(chemokine)和其受器(receptor)間的趨化作用與癌症的轉移有關，而我們先前的研究結果也指出趨化激素19及21（CCL19及CCL21）的受器第七型趨化激素受器(chemokine receptor7, CCR7)在乳癌細胞中會高度表現並在乳癌細胞轉移至淋巴結中扮演著重要的角色。此外，在乳癌病人中，CCR7也與原位腫瘤轉移至淋巴系統有著密切的關係。在本研究中發現TAK1在MDA-MB-231乳癌細胞中會高度活化，當我們使用TAK1 shRNA及TAB1 shRNA去抑制TAK1的訊息路徑後發現CCR7的表現也隨之減少，而在使用TAK1激酶活性抑制劑5Z-7-oxozeaenol（5Z-O）後也發現同樣的現象。而經由Promoter deletion assay結果顯示TAK1調控CCR7的重要位置是落在CCR7轉錄起始點前223到500 bps的位置。進而，利用染色質免疫沉澱（Chromatin immunoprecipitation）分析顯示TAK1是藉由c-Jun及NFκB兩個轉錄因子共同調控CCR7的表現。另外，我們也發現在乳癌組織中p-TAK1和CCR7的表現也呈現正相關性，這與在細胞株中的結果是一致的。

Transforming growth factor-β (TGF-β)-activated kinase 1(TAK1), a member of the MAPKKK family, was identified as a protein kinase that involved in c-Jun N-terminal kinase(JNK)/p38 MAPKs and NF-κB signaling pathways. This kinase also participated in inflammatory and innate immune responses as well as the aggressiveness in several types of cancer. Association with its specific activator protein, TAK1 binding protein1 (TAB1), triggers TAK1 kinase activity. Recent evidences suggest that the interaction between chemokines and their cognate receptors involved in cancer metastasis. Our previous results demonstrated that CC chemokine receptor 7 (CCR7), the receptor for the two major chemokines CCL19 and CCL21, is up-regulated in breast cancer cells and may play an important role in the trafficking and homing of breast cancer cells to lymph nodes. In addition, CCR7 expression is positively correlated with lymphatic metastasis in the primary tumors of patients with breast cancer. Here, we show that TAK1 is constitutively activated in MDA-MB-231 breast cancer cells and suppression of TAK1 signaling by using TAK1 shRNA or TAB1 shRNA significantly reduces expression of CCR7 in these cells. In addition, the TAK1 inhibitor 5Z-7-oxozeaenol, a resorcylic lactone of fungal origin, also inhibited TAK1 kinase activity and CCR7 expression. Promoter deletion assay identified a responsive element localized between -500/-223 bp from transcription start site is critical for TAK1 to activate CCR7 transcription. Furthermore, Chromatin immunoprecipitation assay(ChIP assay)identified two transcription factors, c-Jun and NFκB participate in TAK1 regulating CCR7 expression. In addition, our study in breast tumor tissues indicated that CCR7 was significantly associated with p-TAK1 expression as found in breast cancer cell lines.

縮寫語-------------------------------------5
中文摘要----------------------------------6
英文摘要----------------------------------7
前言----------------------------------------8
研究目的----------------------------------10
實驗材料----------------------------------11
實驗方法及步驟-------------------------14
實驗結果----------------------------------25
討論----------------------------------------41
結論圖表----------------------------------43
Supplementary Data------------------44
附錄----------------------------------------47
參考文獻----------------------------------57

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