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系統識別號 U0026-0812200912054586
論文名稱(中文) 鉀氯離子共同輸送體促進腫瘤轉移的能力
論文名稱(英文) The potential role of potassium chloride cotransporter to increase metastasis
校院名稱 成功大學
系所名稱(中) 微生物及免疫學研究所
系所名稱(英) Department of Microbiology & Immunology
學年度 94
學期 2
出版年 95
研究生(中文) 翁子玉
研究生(英文) Tzu-yu Weng
學號 s4693409
學位類別 碩士
語文別 中文
論文頁數 45頁
口試委員 指導教授-周振陽
指導教授-劉校生
召集委員-戴明泓
中文關鍵字 鉀氯離子共同輸送體  轉移 
英文關鍵字 KCC  metastasis 
學科別分類
中文摘要 鉀氯離子共同輸送體 (KCl cotransporter, KCC) 在子宮頸癌及卵巢癌細胞的生長和侵犯過程中扮演一定重要的角色。在我們先前的研究中發現,KCC3 可以促進腫瘤的生成。因此,在這個研究中,我們想進ㄧ步探討 KCC 是否也可以促進腫瘤的轉移。我們在 NIH/3T3 系列的 7-4 細胞中轉殖 KCC,而 7-4 細胞中有ㄧ個 inducible Ha-rasV12 transgene 的系統可以在老鼠身上造成肺部轉移。我們利用此研究模式,從靜脈注射轉殖 KCC 的 7-4 細胞。研究結果發現,轉殖 7-4-hKCC3 和 7-4-mKCC4 細胞比 vector alone 轉殖的7-4 細胞能在老鼠身上產生更多的肺部轉移,而且似乎 KCC3 具有更強的促進轉移能力。我們後續探討 KCC 促進肺部轉移的可能機轉,實驗發現 KCC3 會增加細胞移行,細胞侵犯及細胞貼附的能力。我們也發現 KCC3 會增加 MMP9 的表現,但則不會增加 MMP2、FAK 397Y 磷酸化及 VEGF的表現。細胞移行、侵犯、貼附能力及 MMP9 的表現上升,都有可能是 KCC3 促進腫瘤轉移的機制,未來希望再進一步探討其中的訊息傳導路徑。
英文摘要 Potassium chloride cotransporter (KCC) is one of the important modulators of growth and invasion in human cervical and ovarian cancer cells. Previously our laboratory found that KCC3 can increase tumor formation in SCID/NOD mice. Thus we investigate whether KCC can enhance tumor metastasis. KCC3 or KCC4 was stably transfected into the NIH/3T3 derived 7-4 cells that contain an inducible Ha-rasV12 transgene, which has been shown to be able to cause metastatic phenotypes in mice. By using this model, we injected these KCC-stable transfected 7-4 cells into the tail veins of the BALB/C mice. The cells with KCC overexpression formed far more aggravated metastasis in the lung compared with the control cells, and no metastatic phenotype was detected in other organs. Our data showed that KCC3 and KCC4 could enhance Ha-rasV12-induced lung metastasis, and KCC3 seemed to play a more potential role in tumor metastasis. We next examined the molecular mechanisms underlying the KCC-enhanced metastasis. The results showed that KCC3 could enhance cell migration, invasion and adhesion. Additionally increased activity of MMP9, but not MMP2, FAK Y397 phosphorylation and VEGF activity were also observed in the 7-4-KCC3 cells. Further effort will focus on revealing the mechanism by which metastasis by KCC3 enhances tumor metastasis.
論文目次 Contents
中文摘要…………………………………………………………………I
Abstract………………………..………………………………………..II
Contents……………………………………………………………..…IV
Figure Index…………………………………………………………….V
Table Index……………………………………………………………..VI
Introduction……………………………………………………………..1
Materials and methods………………………………………………….4
Materials…………………………………………………………………5
Machinery…………………………………………………………..5
Materials for cell culture…………………………………………..5
Drugs………………………………………………………………..6
Antibodies…………………………………………………………..7
Methods………………………………………………………………….8
Cell culture…………………………………………………………8
Western blotting……………………………………………………8
Experimental metastasis assay……………………………………9
Hemotaxylin-Eosin staining……………………………………...10
Migration assay…………………………………………………...10
Transwell invasion assay…………………………………………11
Adesion assay……………………………………………………..11
Gelatin-zymography……………………………………………...12
Mouse VEGF ELISA……………………………………………..13
Buffers………………………………………………………………….14
Results………………………………………………………………….18
KCC3 enhances tumor formation in SCID mice.............………19
KCC increases lung metastasis in the mouse-xenograft model..19
KCC3 can increase cell migration, invasion, and adhesion……21
KCC3 can increase MMP9 secretion but not MMP2 and FAK Y397 phosphorylation…………………………………………….22
KCC3 can not augment VEGF secretion………………………..23
Exogenous Ras Val12 was higher in 7-4-hKCC3 cell……………24
Discussion………………………………………………………………25
References………………………………………………………………28
Figures………………………………………………………………….34
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