研究生: |
陳永富 Yung-Fu Chen |
---|---|
論文名稱: |
Wnt-EGFR訊息傳遞途徑動力學計算研究暨抑制劑效應模擬 Modeling of Kinetics of the Wnt-EGFR Signaling Pathway and Inhibitor Effects on its Kinetic Behaviors |
指導教授: |
孫英傑
Sun, Ying-Chieh |
學位類別: |
碩士 Master |
系所名稱: |
化學系 Department of Chemistry |
論文出版年: | 2012 |
畢業學年度: | 100 |
語文別: | 中文 |
論文頁數: | 91 |
中文關鍵詞: | 訊息傳遞路徑 、抑制劑 、模擬 |
英文關鍵詞: | signaling pathway, inhibitor, modeling |
論文種類: | 學術論文 |
相關次數: | 點閱:87 下載:3 |
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Wnt-EGFR 訊息傳遞途徑是已知與細胞增殖、分化和凋亡有很大的相
關性。在許多癌症裡,發現這些訊息傳遞路徑的異常情形。我們利用電
腦模擬的方法,計算此兩個訊息傳遞路徑的動力學模型,來幫助我們了
解這些訊息傳遞路徑的幾個效應。
以現有的模型為基礎,根據此訊息傳遞途徑的動力學效應,以及相
關的實驗數據來擴充模型。首先,我們增加一條強度合理的負回饋路徑
到兩個不同模型的EGFR 路徑中,當ERKpp 對Raf-1 抑制方程式中K1
值在0.01-10 的區間時,計算結果顯示,此路徑對兩個不同模型中的
ERKpp 濃度有不同效應。路徑中含有Braf 的模型,此負回饋路徑在模型
中無法扮演有效抑制ERKpp 濃度的角色。反之路徑中沒有Braf 的模型,
此負回饋路徑可以有效使ERKpp 濃度表現降低。第二,加入EGFR 與
Wnt 路徑之間的交談(crosstalk)反應路徑,若為正回饋路徑,當β-catenin
無過度表達時,其促使未知因子Y 的濃度大小對於ERKpp 活化濃度扮演
開關效應(switch-like)的行為。若為負回饋路徑,因較強的負回饋效應,
使得Wnt 訊號刺激期間的ERKpp 活化濃度有大幅度震盪的行為,Wnt
訊號結束後ERKpp 濃度迅速降回低點。
iii
此外,我們添加蛋白激酶抑制劑,探討其對磷酸化ERK 表現的效應。
以Wnt 訊號短暫刺激後,以及β-catenin 過度表達造成的ERKpp 濃度失
調時,其濃度被抑制的差異,比較抑制效果。加入單一抑制劑對多個蛋
白激酶同時抑制,其模擬結果顯示,同時對多個蛋白激酶抑制且抑制強
度相同時,因為數個被抑制者會競爭抑制劑的濃度,所以其抑制效果比
單一抑制劑對Raf-1 單獨抑制時差,效應差距最大時,減少的ERKpp 濃
度為單獨抑制Raf-1 時的1/6 倍。單一抑制劑單獨抑制Raf-1 為較佳抑制
效果,可以利用模擬結果探討抑制劑選擇性問題。
加入多個抑制劑時,兩個抑制劑分別抑制Raf-1 及MEK 時,對Raf-1
與MEK的抑制劑兩者濃度比例大於1.5 倍時(濃度總合同單一抑制劑的濃
度時),比起單一抑制劑同時抑制Raf-1 與MEK 且強度相同時,有較好的
抑制效果使ERKpp 表現有明顯的降低。因此兩個抑制劑分別抑制Raf-1
及MEK,且增加Raf-1 抑制劑的濃度時有較佳的抑制效果。
這些結果使我們加深了解兩訊息傳遞路徑,同時對未來多目標蛋白
激酶抑制劑的設計是有幫助的。
The Wnt and EGFR signaling pathways are known to relate to cell proliferation, differentiation, and apoptosis. Deregulation of these signaling
pathways were found in various kinds of cancers. Toward better
understanding of these two pathways, we used computer modeling method to
model kinetics of these pathways.
Based on currently available models, we expanded the model in order to
include more effects in kinetics of these pathways and correlate with available
experimental data. First, we added a negative feedback loop on the EGFR
pathway which has inhibition effect on Raf-1 by ERKpp. When the K1 value
of Raf-1 inhibition reaction was set in the range of 0.01 to 10, the
computations gave that addition of this loop results in two different effects in
the two models we used. The negative feedback loop has a little effect on
ERKpp level in the model which includes Braf. In contrast, the negative
feedback loop makes the level of phosphorylated ERK go down in the model
without Braf. Second, a crosstalk between EGFR and Wnt pathways was
added and kinetic modeling gave that:in the case this is a positive feedback
loop, it induces the switch-like behavior of ERKpp expression by varying the
concentration of the added factor Y when the β -catenin was not
overexpressed. In the case it is a negative feedback loop, due to the stronger
v
negative feedback effect, that during Wnt signal stimulate the concentration of
ERKpp has an oscillation behavior with larger amplitude during the period of
Wnt signal stimulation. After that, concentration of ERKpp falls back to low
level quickly.
In addition, we investigated effect of adding kinase inhibitor(s) on the
level of phosphorylated ERK (ERKpp) under the condition of β-catenin
overexpression plus undergoing a wnt signal transient stimulation. In the case
of adding one kinase inhibitor, the modeling gave that:kinase inhibitor of
multiple-targets of the same strength have less inhibitory effect than inhibitor
of singlet-target of Raf-1 kinase. Reduction of concentration of ERKpp was as
small as to 1/6 fold only compared with the case of singlet-target in the
examined model. Furthermore, we investigate effects of multiple-target
inhibitors inhibiting Raf-1 and MEK kinases. It was found that in the case the
ratio of Raf-1 inhibitor concentration to MEK inhibitor concentration is larger
than 1.5, the inhibitor effect is better than one inhibitor of multiple-target with
the same inhibition strengths. These results deepen our understanding of these
two pathways and should be useful for future multiple-target kinase inhibitor
design.
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