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研究生:吳旭唐
研究生(外文):Syu-Tang Wu
論文名稱:山葵萃取物對肝癌細胞毒性之研究
論文名稱(外文):Study of cytotoxicity of Wasabia japonica extracts on Hepatocellular Carcinoma
指導教授:周芬碧周芬碧引用關係
指導教授(外文):Fen-Pi Chou
學位類別:碩士
校院名稱:中山醫學大學
系所名稱:生化暨生物科技研究所
學門:生命科學學門
學類:生物科技學類
論文種類:學術論文
論文出版年:2013
畢業學年度:101
語文別:中文
論文頁數:61
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食用山葵有多種好處,促進腸胃消化及吸收、抗菌及抗發炎。過去的研究中發現山葵中的成分可以抑制胃癌細胞的增生。在本研究中,發現山葵水萃物對於肝癌Hep 3B 細胞具有毒性。使用DAPI 染色確認細胞死亡的方式,實驗結果顯示出Hep 3B 細胞中具有 DNA 濃染的情形,接著以流式細胞儀Annexin V-Fitc 和PI 染色分析Hep 3B 細胞的細胞周期及死亡方式,發現山葵萃取物濃度增加會使Hep 3B 細胞Sub G1 期增加,證實山葵萃取物使Hep 3B 細胞走向細胞凋亡。使用Western blot 分析細胞凋亡內在路徑及外在路徑中蛋白表現。最後要了解山葵萃取物在動物實驗中對於腫瘤的效果,經餵食山葵萃取物老鼠組別腫瘤大小表現有抑制生長的現象。綜合以上結果,本研究顯示出山葵有抑制肝癌效果,並且未來可將山葵成為治療肝癌的化學預防功能性食品。

Edible of wasabia japonica has many beneficial, promote digestion and absorption, antibacterial and anti-inflammatory. Previous studies found wasabia japonica ingredients can inhibit proliferation in gastric cancer cell. In this study, wasabia japonica found cytotoxic effect in Hep 3B cancer cell. The results experimental Hep 3B cancer cell DNA concentrations of cell death by DAPI staining. The found Hep 3B cell cycle and apoptosis was assessed using flow cytometry Annexin V-Fitc and PI staining. The results make increased Hep 3B cells Sub G1 phase and increased apoptosis quadrant by treated wasabia japonica in added concentration. We confirming Hep 3B cancer cells of occur apoptosis by treated wasabia japonica. Western blotting used to detect extrinsic pathway and intrinsic pathway of apoptosis protein expression. Finally, we used to animal experiments of tumor size in the treated wasabia japonica. The results found inhibit mice tumor growth of feeding wasabia japonica. Collectively, this study shows the inhibit liver cancer of wasabia japonica, and the future can eat wasabia japonica as chemopreventive functional food.

壹、中文摘要-1
貳、英文摘要-2
參、縮寫檢索表-3
肆、緒論-5
一、緒論-5
二、肝臟生理學-5
三、肝癌介紹-6
四、山葵介紹-10
五、細胞凋亡介紹-11
六、實驗蛋白簡介-12
伍、實驗動機-14
陸、研究架構-15
柒、實驗材料與方法-16
一、實驗細胞-16
二、實驗動物-16
三、實驗試劑藥品-16
四、實驗器材-19
五、實驗設備-20
六、實驗方法-21
捌、實驗結果-32
一、山葵萃取物對肝癌Hep 3B細胞毒性分析-32
二、山葵萃取物對肝癌Hep 3B細胞之DAPI細胞螢光染色-32
三、PI 染色觀察山葵萃取物對於Hep 3B細胞週期分布-33
四、Annexin V-Fitc/PI 染色確認山葵萃取物導致Hep 3B產生細胞凋亡-34
五、山葵萃取物刺激肝癌細胞表面接受器活化細胞凋亡外在路徑-35
六、內在路徑caspase 8 蛋白活化促使內在路徑蛋白活化而產生細胞凋亡-36
七、山葵萃取物對打入肝癌Hep 3B細胞之裸鼠動物實驗結果‧‧‧36
八、裸鼠腫瘤蛋白表現結果-37
玖、討論-38
拾、參考文獻-43
拾壹、圖表與說明-52
拾貳、附錄-61


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