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系統識別號 U0026-2707201511330100
論文名稱(中文) 探討以Eps8為標靶之抗大腸癌藥物及其分子機制
論文名稱(英文) Investigation of Therapeutic Drugs Targeting Eps8 for Colorectal Cancer Treatment
校院名稱 成功大學
系所名稱(中) 藥理學研究所
系所名稱(英) Department of Pharmacology
學年度 103
學期 2
出版年 104
研究生(中文) 賴啟楹
研究生(英文) Cremilda Lai
學號 S26025016
學位類別 碩士
語文別 英文
論文頁數 37頁
口試委員 指導教授-呂增宏
口試委員-馬明琪
口試委員-張雋曦
中文關鍵字 大腸直腸癌  Eps8  藥物  SW620  SW480  HT-29 
英文關鍵字 colorectal cancer  Eps8  drugs  SW620  SW480  HT-29 
學科別分類
中文摘要 大腸直腸癌是世界第三大常見的癌症。據過去研究指出,Eps8在大腸直腸癌病人檢體當中有高度的表現量而Eps8的表現量又會造成癌症患者預後較差的情形。Eps8為一種致癌蛋白質,與致癌基因Src及細胞癌化蛋白質FAK有關。因此,找出抑制Eps8表達的試劑有助於大腸癌治療藥物的發展。本研究的目的是要探討Eps8抑製劑的分子機制,通過connectivity map,我們選用了11種與Eps8表達相關的藥物進行測試,最終篩選出M9作為後續探討。利用人類大腸直腸癌細胞株HT-29,SW480和SW620進行細胞實驗,從細胞存活試驗中我們證實M9對於細胞增生有顯著的抑制效果。通過RT-PCR我們驗證了M9抑制Eps8基因轉錄的能力。另外,從western blot的結果發現M9具有濃度依賴性及時間依賴性而抑制Eps8、FAK和Src蛋白質的表達與活性。在流式細胞儀分析中我們觀察到M9會導致大腸癌細胞滯留在G1細胞週期中,影響細胞生長。以上的實驗結果指出M9可以作為Eps8抑製劑並能有效抑制癌細胞的生長,對於發展成為大腸癌用藥具有莫大的潛力。
英文摘要 Colorectal cancer is the third most common cancer in the world. It has been reported that Eps8, epidermal growth factor receptor pathway substrate 8, is expressed at elevated levels in colon cancer and high levels of Eps8 results in poor prognosis of cancer patients. Eps8 acts as an oncoprotein in human cancer. Therefore, an agent that attenuates Eps8 expression can be developed to be therapeutic drug for colon cancer. The aim of this study is to investigate the underlying mechanisms of M9, a potential Eps8 inhibitor for the treatment of colon cancer. Through connectivity map, which includes 6100 drug-mediated expression profiles, 11 candidate drugs were shown to have anti-correlated expression patterns of Eps8. Human colon cancer cell lines Ht-29, SW480 and SW620 were used as model in this study. Results from cell viability assay determined that M9, one of the 11 candidate drugs, inhibited cell proliferation of the treated cells. Then we validated that M9 inhibited eps8 gene transcription through RT-PCR. Moreover, results from western blot analysis revealed that M9 suppressed the expression of Eps8, the activity of FAK and Src and the expression of FAK and Src in dose- and time- dependent manner. In addition, in the flow cytometry analysis, M9 resulted in G1 phase arrest in the colon cancer cell lines and affecting cell proliferation. These findings indicate that M9 is an Eps8 inhibitor and can inhibit cancer cell growth, suggesting its repositioning potential for colon cancer treatment.
論文目次 Abbreviations i
Abstract in English ii
Abstract in Chinese iii
Acknowledgement iv
List of Figures vii
General introduction 1
Colorectal cancer 1
EGFR and the treatment of colorectal cancer 1
Eps8 2
Connectivity Map 4
Specific aims 5
Material and Experimental Procedures 6
Results 11
Discussion 15
References 19
Appendix 32
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