2016-08-012024-05-14https://scholars.lib.ntu.edu.tw/handle/123456789/658808摘要：肺癌為國內最常見的癌症，因此找出肺癌新的治療標的及開發治療策略對國人健康至為重要。Keap1 (Kelch-like ECH-associated protein 1)是一個調控細胞面對氧化壓力反應的關鍵轉錄因子Nrf2 (nuclear factor-erythroid-2-related factor 2)蛋白質穩定與降解的E3 泛素連接酶。由於Nrf2 已知能夠調控許多二期解毒酵素(phase II detoxifyingenzymes) 、藥物運輸蛋白與抗氧化酵素基因例如HO-1、NQO1 與MRP 等基因，因此Keap1 與Nrf2 在腫瘤細胞對化學治療的敏感性與產生抗藥性的機制上已有需多相關研究證實其重要性。然而在調控其他腫瘤惡化進程中重要的細胞功能上就十分缺乏相關的研究報導，我們在初步的實驗中以本土肺癌病患的臨床組織進行Keap1 與Nrf2 的免疫染色分析，發限到Keap1 的蛋白質表現與肺癌患者的臨床期別及淋巴轉移情形有高度的相關性，同時也確認了Keap1 表現量與病患的存活時間呈現高度正相關。此外在細胞實驗中也證實了以RNAi 抑制Keap1 蛋白質表現後，顯著的增加了非小細胞肺癌細胞株的移行與浸襲能力。在此計畫中我們將進一步研究Keap1 與Nrf2 在非小細胞肺癌惡化與轉移過程中的角色，我們將進行一系列細胞與動物實驗來驗證Keap1 與Nrf2在肺癌細胞株中調控細胞移行與轉移的能力，並將試圖找出Nrf2 所調控的新穎基因與Keap1 調控的新穎蛋白質，同時進行這些新穎分子與Keap1 或Nrf2 之間交互作用的分子機制探討。藉由執行此研究計畫我們將對Keap1 與Nrf2 在非小細胞肺癌惡化與轉移過程中的角色有更深入的了解並提供未來發展肺癌治療新穎標靶/策略的立基。<br> Abstract: Kelch-like ECH-associated protein 1 (Keap1) is an E3-ligase participated in the cellulardefense response against oxidative stress through nuclear factor-erythroid-2-related factor 2(Nrf2). Since Nrf2 was known as a key transcriptional regulator of redox-balancing genes(e.g., heme-oxygenase (HO)-1), phase II detoxifying genes (e.g., NADP(P)H quinoneoxidoreductase-1 (NQO1)), and drug transporters (e.g., multidrug resistant proteins (MRPs)),the Keap1-Nrf2 axis was intensively investigated in the chemo-resistance of multiple cancersin previous studies. However, the role of Keap1 in regulating cancer motility is stillcontroversial. Our preliminary data shows that Keap1 expression was decreased in specimensfrom NSCLC patients with lymph node metastasis compared with patients without metastasis.Higher Keap1 expression levels were correlated with the survival of NSCLC patients.Moreover, manipulation of Keap1 expression affected cell migration/invasion abilities. In thisproject, we will investigate the contribution of Keap1-Nrf2 axis in the progression ofnon–small cell lung cancer (NSCLC). A series of in vivo and in vitro assays will be performedto elucidate the contribution of Keap1-Nrf2 axis in lung cancer mobility and progression. Theunderlying molecular mechanism including identifying novel target genes of Keap1-Nrf2 axisand Keap1 E3 ligase substrates will be investigated. Through the complete of this study, wemay obtain new insights into the roles of Keap1-Nrf2 axis in the progression of NSCLC. Wemay also provide useful information for developing therapeutic strategies for NSCLC.Keap1Nrf2非小細胞肺癌細胞浸襲腫瘤轉移Kelch-like ECH-associated protein 1 (Keap1)nuclear factor-erythroid-2-related factor 2 (Nrf2)non–small cell lung cancer (NSCLC)cell mobilitymetastasis