探討內質網壓力在肝癌細胞中誘導Nucleobindin-2表現的機制與角色

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標題:	探討內質網壓力在肝癌細胞中誘導Nucleobindin-2表現的機制與角色 To investigate the mechanism and role of Nucleobindin-2 during endoplasmic reticulum stress in hepatoma cell
作者:	李任豪
貢獻者:	生物科技系洪瑞祥
關鍵字:	糖解作用內質網壓力蛋白質合成即時聚合酶鎖反應 glycolysis ER stress protein synthesis Real-time PCR
日期:	2014
上傳時間:	2015-10-26 20:28:05 (UTC+8)
摘要:	內質網的功能主要為蛋白質合成及修飾，內質網也連接著著細胞核、細胞質和細胞膜。而許多人類疾病都與內質網壓力息息相關，如退化性神經元疾病與糖尿病二型和腫瘤。在過去研究中指出腫瘤可觀察到內質網壓力，而內質網壓力對於腫瘤發展和治療扮演著重要的角色，在先前研究我們證實內質網壓力可誘導 nucleobindin-2 (NUCB-2) 表現。接著在本研究中，我們希望去探討內質網壓力下誘導 nucleobindin-2 (NUCB-2) 的機制及所扮演的角色。首先我們利用了動物實驗模式，探討內質網壓力與 NUCB-2，結果顯示在給予內質網壓力誘導劑時可以明顯誘導 NUCB-2 的表現。此外我們也利用 shRNA 來降低內質網壓力下 NUCB-2 的表現，另外透過即時聚合酶鏈鎖反應分析當 Huh-7 及 Huh-7 NUCB-2 shRNA 細胞處於內質網壓力產生時糖解作用相關基因的表現。結果顯示，當內質網壓力時可以誘導 hexokinase-1 mRNA 的表現，而 Huh-7 NUCB-2 shRNA 細胞則無法誘導 hexokinase-1 mRNA 的表現。未來將進一步探討在內質網壓力下 NUCB-2 如何調控 hexokinase-1表現的詳細機制。 The endoplasmic reticulum (ER) is important organelles in protein synthesis, and connected to the nucleus, cytoplasm and cell membrane in the cell. The ER stress response is suggested to contribute to several types of human disease, including degenerative neuronal disorders and type II diabetes and tumor. In previous studies indicated the tumor was observed in the ER stress, the ER stress in the development and treatment of tumors, plays an important role, In a previous study we demonstrated that ER stress can induce nucleobindin-2 (NUCB-2) expression. In this study, we would like to investigate the mechanisms and role of nucleobindin-2 (NUCB-2) expression in response to ER stress. First of all, induction of NUCB-2 expression by ER stress inducer was observed in animal model. Meanwhile, downregulation of NUCB-2 expression was performed by NUCB-2 shRNA in Huh-7 cells during ER stress. The glycolysis-related genes were investigated by real-time PCR in response to ER stress in Huh-7 and Huh-7 NUCB-2 shRNA cell lines. Increased hexokinase-1 mRNA expression was observed in Huh-7 cells during ER stress. However, inhibition of NUCB-7 expression was found in Huh-7 NUCB-2 shRNA cells in response to ER stress. Taken together, expression of NUCB-2 was increased during ER stress. Furthermore, investigations are required to identify the detail mechanism of NUCB-2 expression and NUCB-2-regulated hexokinase-1 expression in response to ER stress.
關聯:	網際網路公開：2019-08-20，學年度：102，65頁
显示于类别:	[生物科技系(所)] 博碩士論文

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