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標題: | 人類RNA靜默複合體(RISC)組成蛋白質參與癌化進程之功能性探討 Functional Analysis of RCK/p54, a Human RISC Component, in Cancer Progression. |
作者: | Yen-Chun Chen 陳妍君 |
指導教授: | 朱家瑩(Chia-Ying Chu) |
關鍵字: | 轉錄後調控,RCK/p54,轉錄抑制,癌症,惡性化, post-translational repression,RCK/p54,translational repression,cancer,cell migration, |
出版年 : | 2013 |
學位: | 碩士 |
摘要: | RCK/p54是DEAD-box蛋白質的一種,在轉錄後調控(post-transcriptional regulation)基因表現扮演重要角色。在酵母菌、非洲爪蟾、以及果蠅中的研究顯示出RCK/p54的同源基因會參與基因的轉譯抑制(translational repression)進而調控基因表現。在人類miRNA路徑中,RCK/p54會與Ago2及其他蛋白質形成靜默蛋白複合體(miRNA-induced silencing complex),抑制目標RNA轉譯或是將其儲存。在人類大腸癌研究發現RCK/p54大量表現於癌細胞中。除此之外,RCK/p54也高度表現在C型肝炎引起的肝癌細胞中。然而,RCK/p54在癌症中所扮演的角色及其機制皆不甚清楚。為了探討RCK/p54於癌症中的所扮演的角色及其機制,我們利用次世代定序以及定量蛋白質體分析與RCK/p54蛋白質結合的RNA,及降低RCK/p54表現量後所造成的下游RNA及蛋白質表現量變化。研究結果顯示癌細胞的轉移能力會隨著RCK/p54過量表現而隨之提高。此外,藉由比對RCK/p54蛋白質所結合的RNA與蛋白質體的資料,我們發現RCK/p54可能直接經由轉譯抑制調控NUDT21以及GPRC5A兩個基因的蛋白表現。經由次世代定序分析去除RCK/p54時HeLa細胞的基因表現變化圖譜,發現當細胞去除RCK/p54時,MAGEE1的表現量上升,顯示其表現可能間接由RCK/p54調控。本研究顯示細胞轉移程度與RCK/p54表現量成正相關,並且可能是經由調控NUDT21、GPRC5A以及MAGEE1進而影響癌症細胞的惡性化。 RCK/p54 is a member of DEAD-box protein family, and it plays a role in post-transcriptional regulation. Previous studies of RCK/p54 homologues in yeast, Xenopus and Drosophila showed that RCK/p54 participated in regulation of translational repression and moderates RNA stability. RCK/p54 is also involved in miRNA pathway. It interacts with Ago2 in miRNA-induced silencing complex (miRISC), which represses mRNA translation or stores target mRNA in processing body (P-body). Several studies in cancer biology showed that RCK/p54 is highly expressed in tumors. However, the detail mechanism of RCK/p54 in regulation of cancer progression is unrevealed. In this study, we aim to identify the role of RCK/p54 in cancer metastasis. We found out that migration of cancer cell was increased in cells overexpressing RCK/p54. Comparing the deep sequencing result of RCK/p54-associated mRNA and the change of proteomic profile in RCK/p54- depleted cell, we found that RCK/p54 may directly regulate NUDT21 and GPRC5A. MAGEE1 mRNA was increased in RCK/p54-depleted cell, but did not associate with RCK/p54, suggesting that MAGEE1 is a potential downstream target of RCK/p54. These findings suggest that RCK/p54 protein regulates cancer metastasis through various pathways and the potential targets or downstream of RCK/p54 were identified. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/17425 |
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顯示於系所單位: | 動物學研究所 |
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