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研究生:張婉婷
研究生(外文):Wang-Ting
論文名稱:抗菌胜肽Tilapia Hepcidin 1-5對抑制癌症細胞之研究
論文名稱(外文):Study on inhibitory effects of Tilapia Hepcidin 1-5 in cancer cells
指導教授:鄭鈞文鄭鈞文引用關係陳志毅陳志毅引用關係
指導教授(外文):Chun-Wen ChengJyh-Yih Chen
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:2010
畢業學年度:98
語文別:中文
論文頁數:88
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TH1-5 是來自於吳郭魚 (Oreochromis mossambicus) hepcidin 的一段抗菌胜肽。由結果顯示,TH1-5 這一個由22 個胺基酸組成的抗菌胜肽,會特異性的抑制人類纖維肉瘤細胞株HT1080 和人類子宮頸上皮癌細胞株 HeLa 的增生和移動,因此將對降低HT1080 細胞和
HeLa cell 的生長加以研究。利用MTT assasy、AO/EtBr 染色和soft agar assay 的實驗結果顯示,TH1-5 會抑制HT1080 細胞和HeLa 細胞的生長。而以SEM 和TEM 觀察細胞的形態,TH1-5 會將HT1080 細胞和HeLa 細胞之細胞膜破壞和摧毀。在wound healing assay 實驗顯示TH1-5 亦可抑制HT1080 細胞和HeLa 細胞的移動。以流式細胞儀分析,TH1-5 會誘導HT1080 細胞和HeLa 走向細胞凋亡並使細胞停滯在S 期。此外,在real-time PCR 結果,TH 1-5 處理HeLa 細胞會使Bcl-2、Bcl-xL,JNK, IL-6, IL-8, IL-10, IL-12, IL-13, IL-15, CAPN5,CTSG, p53, caspase-3, NFκΒ 和 INF-γ 之 mRNA 相較於沒有處理會高度表現,而在HT1080 細胞會使IL-2, CTSG 和INF-γ 高度表現。在由以上結果顯示TH1-5 對細胞的毒性的機制,將來可成為對抗人類纖維肉瘤和人類子宮頸上皮癌之化學治療藥物試劑。

TH1-5 from antimicrobial peptides of marine organisms, tilapia (Oreochromis mossambicus) hepcidin. The results indicated that TH1-5, a synthetic 22-mer antimicrobial peptide, specifically inhibited human fibrosarcoma cell (HT1080 cell line) and human cervical epithelioid
carcinoma cells (HeLa cell line) proliferation and migration. The way in which TH1-5 inhibited HT1080 and HeLa cell growth was then studied.TH1-5 inhibited HT1080 cell and HeLa cell growth in a concentration-dependent manner according to an MTT analysis, which was confirmed by a soft-agar assay and AO/EtBr staining. Scanning electron microscopy and Transmission Electron Microscope revealed that TH1-5 caused lethal membrane disruption in HT1080 cancer cells and HeLa cells, and a wound healing assay supported that TH1-5 decreased the migration of HT1080 cells and HeLa cells. A flow cytometric analysis,
showed that the TH1-5 induced apoptosis. In addition, Bcl-xL,JNK, IL-6,IL-8, IL-10, IL-12, IL-13, IL-15, CAPN5, CTSG and INF-γ mRNA expression was upregulated after treatment with TH 1-5 in HeLa cell to compared the untreated group. And in HT1080 cell, IL-2, IL-8, CTSG and INF-γ mRNA expression was upregulated. These findings suggest a
mechanism of cytotoxic action of TH1-5 and indicate that TH1-5 may be a promising chemotherapeutic agent against human fibrosarcoma cells and human cervical epithelioid carcinoma cells.

目錄
壹、中英文摘要…………………………………1
貳、序論…………………………………………4
參、研究動機及架構……………………………17
肆、實驗材料……………………………………19
伍、實驗方法……………………………………22
陸、實驗結果……………………………………37
柒、討論…………………………………………44
捌、參考文獻……………………………………48
玖、圖表與說明…………………………………57
拾、附圖表………………………………………84

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