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研究生:林姿吟
研究生(外文):Zi-Yin Lin
論文名稱:OSU-2S誘導口腔鱗狀細胞癌產生細胞凋亡及自噬作用
論文名稱(外文):OSU-2S induces apoptosis and autophagy in human oral squamous cell carcinoma
指導教授:翁靖如
指導教授(外文):Jing-Ru Weng
學位類別:碩士
校院名稱:中國醫藥大學
系所名稱:生物科技學系碩士班
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:2015
畢業學年度:103
語文別:中文
論文頁數:71
中文關鍵詞:口腔癌OSU-2S細胞週期細胞凋亡自噬
外文關鍵詞:Oral cancerOSU-2SCell cycleApoptosisAutophagy
相關次數:
  • 被引用被引用:0
  • 點閱點閱:130
  • 評分評分:
  • 下載下載:5
  • 收藏至我的研究室書目清單書目收藏:0
口腔癌是癌症十大死亡原因之一,從1991 年起,在台灣男性口腔癌的死亡人數持續上升。有研究報告顯示罹患口腔癌的風險因素,包括:喝酒、嚼食檳榔、口腔衛生差,與人類乳頭狀瘤病毒感染和口腔癌有密切相關。
OSU-2S 由FTY720 一個免疫抑制劑化學衍生而來的化合物;已被文獻證明不具免疫抑制的活性卻擁有抑制動物肝臟腫瘤的生長。在本論文中,我們想測試OSU-2S 是否能抑制口腔癌細胞的生長。以MTT assay 檢測細胞存活率,OSU-2S 顯示能抑制口腔癌(SCC-4)細胞的生長,IC50 值為2.9 microM。流式細胞儀分析析發現OSU-2S 使細胞週期停滯於G0/G1 期,並促使凋亡細胞增加的比例由9.1 %上升到92.7 %。此外,西方墨點法分析與顯微鏡觀察到OSU-2S 會負面調控Akt及ERK的磷酸化,且增加自噬分子LC3B-II 的表現增加和自噬小體的出現。以上實驗顯示,OSU-2S 具有抑制口腔癌細胞(SCC-4)生長,使細胞停滯在G0/G1 期,且藉由誘發凋亡與自噬。

Oral cancer has been one of the top 10 causes of death for cancer since 1991 in Taiwan, and the death toll population of oral cancer in males has been increasing. The risk factors have been reported to be closely associated with oral cancers including alcoholic consumption, betel quid chewing, poor oral health, and human papilloma virus infection.
OSU-2S is developed from FTY720 analogue which is a immunosuppressive agent, but it has been shown to have non-immunosuppressive activity, and has highly effective to suppressed hepatocellular carcinoma tumor growth in vivo. In this study, we investigated that OSU-2S affected cell growth in oral cancer cells. The cell viability of OSU-2S was assessed by MTT assay, and the IC50 value is 2.9 microM in SCC-4 cells. Results from flow cytomtric analysis indicated that OSU-2S led to G0/G1 arrest of the cell-cycle and increased apoptotic cell from 9.1 % to 92.7 %. In addition, OSU-2S down-regulated the phosphorylation of Akt and ERK and increased the expression of LC3B-II by western blotting. OSU-2S also increased accumulation of autophagosome. The data was suggested that OSU-2S suppressed cell growth and cause G1 phase arrest, subsequently resulting to cell apoptosis and autophagy in oral cancer cells.


總目錄 I
圖目錄 IV
表目錄 VI
縮寫目錄表 VII
中文摘要 IX
英文摘要 X
第一章 研究緣起 1
第二章 文獻回顧 3
第三章 研究材料與方法 6
第一節 實驗材料 6
一、細胞株 6
二、實驗藥物 6
三、藥品試劑 6
四、儀器設備 10
第二節 實驗方法 12
一、實驗流程 12
二、實驗藥品配置 13
三、細胞培養 14
(一)、配置細胞培養基 14
(二)、細胞株培養條件 14
(三)、解凍細胞 14
(四)、冷凍細胞 15
四、細胞存活率試驗(MTT assay) 16
五、流式細胞儀分析(Flow cytometry) 18
(一)、細胞週期檢測 18
(二)、細胞凋亡檢測 20
(三)、活性氧化物質的測定 21
六、細胞核濃縮現象(DAPI assay) 23
七、酸性小體檢測(Acridine orange staining) 24
八、西方墨點法(Western blotting) 26
(一)、細胞總蛋白質的抽取 26
(二)、蛋白質濃度測定 26
九、統計分析 32
第四章實驗結果 33
第一節OSU-2S對口腔細胞癌細胞(SCC-4、SCC- 33
2095、Ca-922)的存活率探討
第二節OSU-2S對口腔癌細胞(SCC-4)細胞週期 34
及相關蛋白質表現量之探討
第三節OSU-2S對口腔癌細胞(SCC-4) DNA受損 36
之探討
第四節OSU-2S對口腔癌細胞(SCC-4) 凋亡影響 37
及相關蛋白質表現量之探討
第五節OSU-2S 對口腔癌細胞(SCC-4) 所引發的 39
活性氧化物質之探討
第六節OSU-2S 對口腔癌細胞(SCC-4)自噬作用 40
及相關蛋白質表現之探討
第七節 實驗結果 42
第五章 實驗討論 58
第六章 結論 62
第七章 參考文獻 64


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