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研究生:陳玲玲
研究生(外文):Ling-Ling Chen
論文名稱:FormosaninC抗大腸直腸癌之作用機轉
論文名稱(外文):The Anti-tumor Mechanism of Formosanin C
指導教授:翁舷誌李政昌李政昌引用關係
指導教授(外文):Shen-Jeu WonJeng-Chang Lee
學位類別:碩士
校院名稱:國立成功大學
系所名稱:微生物暨免疫學研究所
學門:生命科學學門
學類:微生物學類
論文種類:學術論文
論文出版年:2003
畢業學年度:91
語文別:中文
論文頁數:86
中文關鍵詞:粒線體細胞凋亡
外文關鍵詞:caspaseapoptosis
相關次數:
  • 被引用被引用:3
  • 點閱點閱:212
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  • 收藏至我的研究室書目清單書目收藏:1
Formosanin C (PF-C)是從傳統中草藥植物Paris formosana Hayata 純化出之天然成份,可誘發大腸直腸癌細胞株HT-29 產生細胞凋亡(apoptosis)之反應。以PF-C 處理HT-29 細胞6 天,其50%之抑制濃度(IC50)為1.04 µM,而以PF-C處理人類週邊單核細胞(PBMC)與人類臍靜脈內皮細胞(HUVEC),其IC50 分別為24 µM 與12.6 µM。以PF-C 處理HT-29 細胞後,細胞膜內面之磷脂絲胺酸(phosphatidylserine)翻轉至細胞外表面,且細胞週期sub-G1 之DNA 含量增加,並觀察到DNA 片段化(DNA fragmentation)之現象,這些結果顯示PF-C 是透過細胞凋亡的過程抑制HT-29 細胞生長。以西方墨點法(Western blot analysis)探討PF-C 誘導細胞凋亡的分子機制,發現會造成caspase-2 之活化、粒線體之Bax 和Bak 表現量增加,以及粒線體之Bcl-XL 表現量減少。另外,以PF-C 處理HT-29 細胞後,會造成粒線體膜電位(ΔΨm)之喪失,cytochrome c 與Smac/DIABLO 由粒線體膜間區釋放至細胞質,XIAP 蛋白質表現量減少並造成caspase-9 與caspase-3 之活化,進而切割活化DFF45,最後誘發DNA 的斷裂。由流式細胞儀與共軛焦顯微鏡分析,發現PF-C 引發細胞凋亡之作用與造成粒線體ΔΨm 喪失之情形可被caspase-2 之抑制劑Z-VDVAD-fmk、caspase-9 之抑制劑Z-LEHD-fmk 和cyclosporin A 所回復。綜觀以上實驗結果推測,以PF-C 處理大腸直腸癌細胞HT-29 後,會造成caspase-2 之活化、粒線體ΔΨm 之喪失、cytochrome c 與Smac/DIABLO 由粒線體釋放至細胞質、XIAP 蛋白質表現量減少,繼而活化下游分子,最終造成細胞凋亡。本篇研究結果針對PF-C 作用分子機制之探討,有助於新藥之設計,以減緩大腸直腸癌疾病之進程並嘉惠病患。
The natural product of formosanin C (PF-C), isolated from Paris formosana Hayata, a traditional Chinese herbal remedy, induces strong apoptotic responses of human colorectal cancer cell HT29. The 50% inhibition concentration (IC50) of PF-C on HT29 cells was 1.04 µM as compared to normal human peripheral blood
mononuclear cells (24 µM) or HUVEC (12.6 µM) at 6 days of post-treatment. The apoptotic process is demonstrated by the phenomena of phosphatidylserine exposure, magnifying of sub-G1 content, and DNA fragmentation. The molecular mechanism events of PF-C triggered apoptosis are associated with the activation of caspase-2, up expressions of mitochondrial Bax and Bak, and down expression of mitochondrial Bcl-XL. Furthermore, treatment with PF-C leads to the loss of mitochondrial membrane potential (ΔΨm), releases of cytochrome c and Smac/DIABLO from
mitochondria to cytosol, decrease of XIAP protein and activations of caspase-9 and -3, and in turn to cleave PARP and DFF45, and then induces DNA fragmentation. The apoptosis and the loss ofΔΨm induced by PF-C are reversed by Z-VDVAD-fmk (caspase-2 inhibitor), Z-LEHD-fmk (caspase-9 inhibitor) and cyclosporin A using flow cytometry or confocal microscopy analysis. Thus, PF-C treatment of colorectal cancer cell HT29 causes activation of caspase-2, loss ofΔΨm and the down stream
apoptotic events to lead the cell death. These results suggest that the molecular mechanism of PF-C may shed light on novel drugs design against carcinogenesis of human colorectal cancer progression.
總 目 錄
中文摘要…………………………………………………II
英文摘要…………………………………………………III
圖表目錄…………………………………………………IV
縮寫對照表………………………………………………VI
誌謝………………………………………………………VIII
緒論……………………………………………………………1
材料與方法……………………………………………………13
結果……………………………………………………………28
討論……………………………………………………………37
參考文獻………………………………………………………45
附圖表…………………………………………………………59
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