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研究生:陳柏翰
研究生(外文):Po-Han Chen
論文名稱:人類類Ste20蛋白激酶Mst4生物功能之研究
論文名稱(外文):The research on the biological functions of a novel human Ste20-like protein kinase Mst4
指導教授:袁俊傑袁俊傑引用關係
學位類別:碩士
校院名稱:國立交通大學
系所名稱:生物科技研究所
學門:生命科學學門
學類:生物科技學類
論文種類:學術論文
論文出版年:2003
畢業學年度:92
語文別:中文
論文頁數:57
中文關鍵詞:蛋白激酶胚胎中心激酶細胞凋亡
外文關鍵詞:Mst4ste20JNKp38apoptosisHEK293MCF-7
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Mst4是屬於人類類Ste20蛋白激酶(human Ste20-like kinase)中的胚胎中心激酶家族(germinal center kinase),其蛋白質大小為46.5 kDa。雖然Mst4和Mst3的N端激酶區域彼此的相似度達到88%,但目前已知Mst4的生物功能卻具有爭議性。先前研究已然發現當Mst4在細胞中大量表現後會造成細胞的生長或死亡。這兩種截然不同的結果。為了釐清Mst4所扮演的角色並了解他的分子機轉我們設計了一些Mst4的突變株並觀察他們在MCF-7及HEK293這兩種細胞中的表現。在我們的實驗中,我們觀察到Mst4會造成MCF-7及HEK293細胞的死亡。經由TUNEL assay的結果,我們更驗證了經由Mst4的誘發可導致細胞產生DNA fragment的現象,這是一種細胞凋亡(apoptosis)的特徵。我們也發現了EGFP-tag會消減Mst4引起細胞凋亡的活性,這與我們實驗室之前所觀察到的現象相吻合。接著我們研究MAPK cascades在Mst4造成MCF-7及HEK293細胞凋亡中所扮演的角色。我們發現了Mst4在MCF-7及HEK293可能會經由不同的路徑造成細胞凋亡。在MCF-7細胞中,Mst4會經由活化JNK的路徑造成細胞的凋亡。但在HEK293細胞中Mst4則可能經由p38 MAPK的路徑導致細胞凋亡。至於這部分詳細的分子機轉,仍需要繼續研究加以解釋。

Mst4 is a novel member of the germinal center kinase subfamily of human Ste20-like kinase with a molecular mass of around 46.5 kDa. Although the kinase domain of Mst4 shares about 88% amino acid sequence homology with that of Mst3, the biological
function of Mst4 is controversial. Previous reports have shown that the overexpression of Mst4 in cells might lead to transformation, growth induction, or contrary, cell apoptosis. To clearify the role of Mst4 and to understand its molecular mechanism in cells, we generated several mutants of Mst4 and study their effect in two cell lines, human breast adenocarcinoma cell line MCF-7 and human embryonic kidney cell line HEK293. Interestingly, we found that Mst4 could induce cell death of MCF-7 and HEK293. By using TUNEL assay, we further demonstrated that Mst4 overexpressed cells exhibited DNA fragmentation, a characteristics of apoptosis. We also found that EGFP-tag may hinder the apoptotic activity of Mst4, which is cinsistent to the previous studies in our laboratory. Subsequently, we studied the role of MAPK cascades in the Mst4-induced apoptosis in MCF-7 and HEK293 cells. Interestingly, we found that Mst4 may induce apoptosis via different MAPK cascades in different cell lines. In MCF-7, a breast tumor cell line, Mst4 may induce apoptosis through JNK pathway, while p38 MAPK pathway may be activated in the Mst4-induced HEK293 apoptosis. The molecular mechanisms underlying these processes, however, are remained to be elucidated.

Abbreviation
1. Introduction 1
2. Materials and methods 7
3. Result 15
4. Discussion 20
5. Reference 23
Appendix 52

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