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標題: | 利用化學蛋白體學鑑定hnRNPA1為槲黃素之目標蛋白與其對攝護腺腫瘤的抗癌機轉研究 Chemical proteomics identifies hnRNP A1 as the molecular target of Quercetin in its anti-cancer effects in PC-3 cells |
作者: | Chia-Chen Ko 柯家錚 |
指導教授: | 周綠蘋(Lu-Ping Chow) |
關鍵字: | 槲黃素,hnRNPA1,化學蛋白體學,攝護腺癌,cIAP1, Quercetin,hnRNPA1,chemical proteomics,molecular target,prostate cancer,cIAP1, |
出版年 : | 2014 |
學位: | 博士 |
摘要: | 槲黃素是一種常見於蔬果中的植物性生物黃酮,已經被用於預防慢性攝護腺炎和攝護腺癌病患,它被報導具有抗發炎、抗氧化和抗癌的作用,但其詳細的抗癌作用機轉並不被完全了解。藉由化學蛋白體學,結合了藥物親和層析和質譜儀的生物資訊分析來研究槲黃素的直接結合蛋白質,我們鑑定到hnRNPA1和槲黃素有交互作用,並由西方點墨法和表面電漿共振生物感測儀確定hnRNPA1與槲黃素交互作用的專一性。並更進一步,利用基因工程將hnRNPA1片段化並分別與槲黃素進行結合反應,我們發現槲黃素和hnRNPA1結合在C-terminal區域,阻礙了hnRNPA1往返細胞核質的正常功能,造成hnRNPA1 堆積在細胞質中,並暫存在stress granules,無法進行正常功能下抗凋亡蛋白質cIAP1 mRNA的轉譯,使得細胞最終走向凋亡。這是第一篇報導hnRNPA1是槲黃素的直接作用蛋白質,並闡釋了槲黃素的抗攝護腺癌機轉之一是透過結合hnRNPA1,影響了細胞內抗凋亡蛋白質的平衡造成癌細胞的死亡。 Quercetin, a flavonoid abundantly present in plants, is widely used as a phytonutrient in prostatitis and prostate cancer. Although quercetin has been reported to have a number of beneficial effects, the cellular target(s) responsible for its anti-cancer action has not yet been clearly elucidated. Here, employing affinity chromatography and mass spectrometry, we identified heterogeneous nuclear ribonucleoprotein (hnRNP) A1 as a direct target of quercetin. A specific interaction between quercetin and hnRNPA1 was validated by immunoblotting and surface plasmon resonance experiments. We found that quercetin bound the C-terminal region of hnRNPA1, impairing the ability of hnRNPA1 to shuttle between the nucleus and cytoplasm and ultimately resulting in its cytoplasmic retention. In addition, hnRNPA1 was recruited to stress granules (SGs) after treatment of cells with quercetin for up to 48 hours, and the levels of cellular inhibitor of apoptosis protein-1 (cIAP1), an IRES-translation dependent protein, were reduced by hnRNPA1 regulation. This is the first report that anti-cancer effects of quercetin are mediated, in part, by impairing functions of hnRNPA1—insights that were obtained using a chemical proteomics strategy. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/55427 |
全文授權: | 有償授權 |
顯示於系所單位: | 生物化學暨分子生物學科研究所 |
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