臺灣博碩士論文加值系統

Naphtho[1,2- b]furan-4,5-dione (NFD)可經由chloroacetaldehyde與2-hydroxy-1,4-naphthoquinone 一鍋合成反應而成，先前的研究發現具備些許的抗癌活性。然而，抑制癌細胞增生的分子機制尚未被探討。本研究以流式細胞儀分析經NFD處理之人類口腔癌細胞株(Ca9-22 cells)，結果顯示細胞週期停滯在G2/M期，進一部探討細胞週期蛋白質的變化，發現cyclin A、CDK1 以及CDK2都有減少的趨勢，及扮演細胞週期抑制者的p27增加。本研究亦發現NFD誘導凋亡使得促進細胞凋亡蛋白Bax、Bad增加，抗細胞凋亡蛋白Bcl-XL、Bcl-2、Mcl-1、XIAP降低，造成cytochrome c的釋出以及capsase-9、capsase-3的活化。

進一步的探討其訊息傳遞，發現NFD會抑制Src的磷酸化及其下游的PI3K和Akt的活化，Ca9-22細胞同時處理NFD與PP2(Src專一性抑制劑)會發現更明顯的細胞週期停滯現象與細胞凋亡的產生。綜合以上實驗結果顯示NFD會抑制人類口腔癌細胞株(Ca9-22)細胞週期停滯於G2/M期，並有誘導細胞凋亡的現象；NFD會抑制細胞增生是經由抑制Src的磷酸化及其下游的PI3K和Akt路徑。所以NFD是一具有潛力抑制人類口腔癌細胞株(Ca9-22)的化合物。

Naphtho[1,2-b]furan-4,5-dione(NFD),prepared from 2-hydroxy-1,4
-naphthoquinone and chloroacetaldehyde in an efficient one-pot reaction, exhibits anti-carcinogenic effects. However, the molecular mechanisms on the anti-proliferative activity in cancer cells have been poorly elucidated. NFD exerted anti-proliferative activity with the G2/M cell cycle arrest and apoptosis in Ca9-22 cells. The NFD-induced cell cycle arrest was correlated with a marked decrease in the expression levels of cyclin A, and their activating partner cyclin-dependent kinases (CDK) 1and 2 with concomitant induction of p27. NFD-induced apoptosis was accompanied with up-regulation of Bax and Bad, and down-regulation of Bcl-2, Bcl-XL, Mcl-1, and X-linked inhibitor of apoptosis (XIAP), resulting in cytochrome c release and sequential activation of caspase-9 and caspase-3.

In the analysis of signal transduction pathway, NFD substantially reduced the phosphorylation of Src and activation of phosphatidylinositol 3-kinase (PI3K) and Akt. The combined treatment of NFD with PP2 (a specific Src inhibitor) significantly enhanced the cell cycle arrest and apoptosis in Ca9-22 cells. These findings suggest that NFD-mediated cytotoxicity of Ca9-22 cells related with the G2/M cell cycle arrest and apoptosis via inactivation of Src and PI3K/Akt-mediated signaling pathways. Thus, NFD appears to be a potential therapeutic agent for killing Ca9-22 cells.

頁碼
中文摘要 2
英文摘要 3
緒論 4
實驗材料與方法 16
結果 23
討論 29
參考文獻 33
圖表說明 44

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