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標題: 高良薑素誘導鼻咽癌細胞p53非依賴性細胞週期S期停滯及細胞凋亡藉由抑制PI3K-AKT訊息路徑
Galangin induces p53-independent S-phase arrest and apoptosis in human nasopharyngeal carcinoma cells through inhibiting PI3K-AKT signaling pathway
作者: 李權峻
Chuan-Chun Lee
關鍵字: 凋亡;Apoptosis
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高良薑素 (galangin: 3,5,7-trihydroxyflavone) 抗癌活性在許多不同種類的癌症皆被證明,然而高良薑素對人類鼻咽癌細胞的影響在目前仍是未被探討的。於研究中發現高良薑素誘導鼻咽癌細胞凋亡的特徵包括: 細胞核濃縮,活化caspae-3及poly ADP-ribose polymerase (PARP) 蛋白質,DNA 斷裂,並誘導鼻咽癌細胞的細胞週期停滯於S 期。在PI3K抑制劑 (LY294002),AKT抑制劑 (MK-2206),細胞內大量表現野生型 p85 alpha 或突變型p85 alpha (缺乏與p110 alpha 結合能力) 基因實驗中,證明高良薑素藉由抑制PI3K-AKT訊息路徑來誘導鼻咽癌細胞凋亡及細胞週期停滯於S期。利用短片段髮夾結構RNA p53 shRNA 抑制鼻咽癌細胞內p53 蛋白質的表現,發現不影響高良薑素的抗癌活性。在ATP競爭試驗及薄層層析法分離試驗中證明高良薑素可藉由與ATP競爭p110 alpha 上的ATP結合位來抑制PI3K活性。由此證明高良薑素也許能當作一種ATP競爭的抑制劑,藉由與ATP競爭PI3K p110 alpha 上的ATP結合位來抑制PI3K-AKT訊息路徑,進而誘導與p53蛋白質表現無關的鼻咽癌細胞凋亡及細胞週期停滯於S期。

Anti-cancer activity of 3,5,7-trihydroxyflavone (galangin) has been documented in a variety of cancer types; however, its effect on human nasopharyngeal carcinoma (NPC) cells remains undetermined. Herein, we show that galangin induces apoptosis of NPC cells, characterized by nuclear condensation, cleavage of pro-caspase-3 and poly ADP-ribose polymerase (PARP), and DNA fragmentation. Treatment with galangin also resulted in an S-phase cell cycle accumulation. Using the phosphatidylinositol 3-kinase (PI3K) inhibitor LY294002, protein kinase B (AKT) inhibitor MK-2206, and ectopic expression of wild type p85 alpha or p85 alpha mutant lacking p110 alpha-binding ability, we show that galangin induces apoptosis and S-phase arrest by attenuating the PI3K-AKT signaling pathway. Short hairpin RNA-mediated silencing of p53 does not block the anti-cancer activity of galangin. ATP-competitive inhibition assay and thin-layer chromatography assay show that galangin can inhibit PI3 kinase activity by competing ATP binding site of p110 alpha. Collectively, galangin might be an ATP-competitive inhibitor that induces apoptosis and S-phase arrest of NPC cells by attenuating the PI3K-AKT signaling pathway in a p53-independent manner.
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