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http://140.128.103.80:8080/handle/310901/458
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| Title: | 合成Id1蛋白質之胜肽拮抗物暨應用表面膜漿共振技術及西方墨點轉漬法研發評估抗癌功效的新方法 |
| Other Titles: | Synthesis of a peptide antagonist of Id1 protein and development of novel methods for evaluation of its anticancer effects using surface plasmon resonance technology and Western blotting |
| Authors: | 廖志偉
Liao, Chih-Wei |
| Contributors: | 龍鳳娣
Lung, Feng-Di
東海大學化學系 |
| Keywords: | Id1蛋白質、Bcl-2蛋白質、Bax蛋白質、β-actin蛋白質、 胜肽拮抗物、peptide 3C、人類乳癌細胞、細胞凋亡、西方墨點轉漬法、表面膜漿共振技術。
Id1 protein, Bcl-2, Bax, β-actin, peptide antagonist, human breast cancer cells, apoptosis, Western blot, surface plasmon resonance. |
| Date: | 2010 |
| Issue Date: | 2011-01-03T05:41:31Z (UTC)
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| Abstract: | Id1 蛋白質對於腫瘤細胞的形成與移動是重要的影響因子之ㄧ。本實驗室的研究目的為抑制Id1 蛋白質的活性以期減緩或抑制癌細胞生長與腫瘤細胞的轉移性。2010年本實驗室的發表論文中以MyoD蛋白質為藍本設計出Id1蛋白質之胜肽拮抗物peptide 3C，發現此胜肽在低濃度下，對癌細胞具有不錯的毒殺效果。所以本論文的研究目的為希望藉由探討癌細胞內特定蛋白質的含量變化，進一步瞭解癌細胞的凋亡與特定蛋白質間的關係。實驗先將人類乳癌細胞(MCF-7 cells)以peptide 3C處理24小時及48小時後，萃取癌細胞內的總體蛋白質並測量其總蛋白質濃度後，依序以西方墨點轉漬法(Western blot)和表面膜漿共振技術(surface plasmon resonance, SPR)偵測癌細胞內特定蛋白質的含量變化。實驗結果發現，兩種對細胞內特定蛋白質定量的方式之結果具有相同的趨勢性：將peptide 3C加入人類乳癌細胞後，會使癌細胞內的Id1 蛋白質和Bcl-2蛋白質含量下降；Bax蛋白質含量上升；β-actin蛋白質含量則是不變。互相比較兩種細胞內特定蛋白質定量的方式之結果，有助於我們研發並評估抗癌功效的新方法。
Id1 protein is an important factor of tumorigenesis and metastasis. The objective of our study is to suppress the activity of Id1 protein in order to slow down or inhibit the tumorigenesis and metastasis. Our recent study reported in 2010 revealed that peptide 3C, a peptide fragment of MyoD protein, exhibited cytotoxic effect on cancer cells at low concentration. The objective of this thesis is to investigate the relationship between apoptosis and the expression of specific proteins by analyzing the changes of expression levels of specific proteins in cancer cells and peptide-treated cancer cells. Human breast cancer cells (MCF-7 cells) were cultured and treated with peptide 3C for 24 hours and 48 hours, followed by extraction of proteins from cancer cells and measuring the total concentration of cellular proteins. Changes of amounts of specific proteins in cancer cells were analyzed by Western blot and surface plasmon resonance technology. Results obtained by these two methods are correlated, indicating that the changes of expression levels of specific proteins in cancer cells are similar：After MCF-7 cells were treated with peptide 3C, the concentrations of Id1 protein and Bcl-2 protein were decreased, the concentration of Bax protein was increased, and the concentration of β-actin protein remained the same as expected. Comparison of these two methods for quantification of concentrations of specific proteins in cancer cells is useful for the development of novel methods for evaluation of anticancer effects of peptides or drugs. Keywords：Id1 protein, Bcl-2, Bax, β-actin, peptide antagonist, human breast cancer cells, apoptosis, Western blot, surface plasmon resonance. |
| Appears in Collections: | [化學系所] 碩博士論文
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| 098THU00065019-007.pdf | | 1975Kb | Adobe PDF | 922 | View/Open | | 098THU00065019-008.pdf | | 316Kb | Adobe PDF | 263 | View/Open |
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