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|Title: ||研究Mst3(mammalian ste20 kinase 3) 在腎臟分枝管狀形成過程的作用|
The Role of Mst3( Mammalian Ste20 Kinase 3) in Renal Tubulogenesis
|Issue Date: ||2010-09-23 13:56:36 (UTC+8)|
|Abstract: ||腎臟發育時，分枝管狀形成的受損，是幼童常見的腎臟衰竭的原因。分枝管狀之形成與細胞之爬行，凋亡及生長週期有關。以HGF 刺激腎臟細胞MDCK，可使其發育成分枝的管狀。當HGF 刺激MDCK，COX-2 蛋白質的表現明顯上升。在缺乏COX-2 的老鼠中，腎臟的分枝過程會受損。另外，腎臟受傷復原時，HGF 接受器，COX-2 以及fibronectin 均會上升。若MDCK 不分泌fibronectin 則管狀分枝無法形成。屬Ste 20 激酶家族的MST3，其蛋白的功能被報導 (1)可促進細胞凋亡 (2) MST3 能控制生長週期 (3) MST3 能抑制細胞移動。這3 項功能均與分化的過程有關。加上我們發現 (i) 在老鼠出生後的腎臟中，MST3 活性上升。(ii) MST3 只有表現在腎臟的細上升枝。(iii) MDCK 大量表現MST3 時， COX-2 明顯上升。 (iv) 必須將細胞培養在舖有fibronectin 之培養皿上，當MDCK 細胞表現大量 MST3 時，才會使細胞移動速度變慢，顯示MST3 與fibronectin 有關。故我們選擇研究MST3 對腎臟發育管狀分枝形成之影響。 In vivo，我們將比較表現活化態MST3 的轉殖性老鼠與野生型MST3 的老鼠，找出MST3 在不同發育時期，在分枝管狀形成的過程，是否有所改變。In vitro，我們將會利用大量表現野生型MST3 及dominant negative MST3 之MDCK 細胞株，研究MST3 是否可經由COX-2 上升，再配合MST3 可增加fibronectin 的分泌而使而促進腎臟分枝發育。再回到in vivo 中，解出MST3 否是經由COX-2 及 fibronectin，控制腎臟分枝管狀發育的詳細機轉。
Disruption of renal branching tubulogenesis during kidney development resulted in renal dysplasia, the major cause of renal failure in young children. The morphogenetic steps are controlled by apoptosis, cell cycle and cell motility. An in vitro model in which HGF induces branching morphogenesis in MDCK epithelial cell culture in three-dimensional collagen gels is established. The COX-2 mRNA and protein expression in MDCK cells are apparently increased after stimulation with HGF. Mouse mutation of COX-2 gene resulted in defects in renal morphogenesis. Besides HGF and COX-2, the interaction of extracelluar matrix (ECM) and cells was also the regulator of branching tubulogenesis. COX-2, HGF receptor, and fibronectin are all increased when mouse was administrated with renal injury. The secretion of fibronectin by MDCK cells promoted renal tubulogenesis. Without the secretion of fibronectin, the tubule could not be formed. The Ste20 family consists of MST1-4 (mammalian Ste20 kinase 1-4)。It was reported that (1) MST3 can promote apoptosis. (2) MST3 can regulate cell cycle and (3) MST3 can inhibit migration. Our results showed (i) the activity of MST3 was elevated in the kidney of 1-2 week old rat (ii) MST3 was exclusively expressed at the thin ascending limb of the kidney. (iii) COX-2 was upregulated in MST3 overexpressed MDCK. (iv) The migration inhibition function of MST3 was appeared when fibronectin was served as the matrix. In vivo, we will compare the active HA-MST3 expressing transgenic mice with wild type MST3 mice to find the effect of MST3 on renal tubulogenesis. In vitro, we will proof that MST3 can stimulate COX-2 to control HGF-induced renal tubulogenesis. Furthermore, HA-MST3 MDCK and dominant negative MST3 will be used to answer if MST3 can secrete fibronectin to lead the formation of renal tubulogenesis. Finally, in vivo and in vitro evidences regarding the relationship between COX-2, fibronectin and MST3 will be considered, rendering new insight into regulating roles for MST3 in renal tubulogenesis.
|Appears in Collections:||[醫學檢驗生物技術系碩士班] 研究計劃|
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