微小核醣核酸與C型肝炎病毒基因體的交互作用與功能性探討

CMUR > College of Medicine > Department of Medical Laboratory Science and Biotechnology > Research reports > Item 310903500/44329

Please use this identifier to cite or link to this item: http://ir.cmu.edu.tw/ir/handle/310903500/44329

Title:	微小核醣核酸與C型肝炎病毒基因體的交互作用與功能性探討
Authors:	鄭如茜(Ju Chien Cheng);黃憲達
Contributors:	健康照護學院醫學檢驗生物技術學系
Date:	2012-07-31
Issue Date:	2012-06-15 11:28:55 (UTC+8)
Abstract:	C 型肝炎病毒感染可造成肝硬化甚至肝癌，為全球性的重要議題。以干擾素結合 Ribavirin 為目前的標準治療策略，然而約有一半的患者並不能被完全根治。因此, 若能抑制調控C 型肝炎病毒感染的宿主細胞因子，將可發展成為新的預防或治療策略。除了宿主蛋白外，微小核醣核酸已被發現可以調控C 型肝炎病毒感染，也因此提供了另一個可行的抗C 型肝炎感染的宿主標的。最近，我們以生物資訊分析肝臟表達較高的微小核醣核酸發現除了已知可以調控C 型肝炎病毒感染的miR-122 外，微小核醣核酸 let-7b 與 miR-16 有機會可與C 型肝炎病毒基因體結合。我們進一步以C 型肝炎病毒複製細胞進行實驗，發現微小核醣核酸let-7b 可以抑制C 型肝炎病毒 RNA 含量，證實我們在生物資訊上的預測結果，並暗示微小核醣核酸let-7b 可能是C 型肝炎病毒感染的負調控因子。在這個計畫裡，我們希望能全面分析經由生物資訊預測可能調控C 型肝炎病毒感染的微小核醣核酸。在計畫第一年，我們將以不同的C 型肝炎病毒複製細胞及感染性病毒顆粒確認這兩個微小核醣核酸調控C 型肝炎病毒的現象。在計畫第二年，我們將進一步瞭解這個交互作用的分子機制。首先，我們將以免疫沉澱法證明微小核醣核酸確與 HCV 基因體共存於RISC 複合體中; 同時，將以突變試驗進一步確認微小核醣核酸與 C 型肝炎病毒基因體的交互序列。第三年，我們將進一步探討微小核醣核酸做為感染C 型肝炎病毒輔助療法的可能性。藉由這個計畫的執行，我們希望能對於新發現可與C 型肝炎病毒基因體序列交互反應的的微小核醣核酸有更清楚的瞭解，並對宿主細胞因子參與 C 型肝炎病毒致病機制提出新的見解。 Hepatitis C virus (HCV) infection is a global health problem and could lead to severe sequels such as liver cirrhosis and hepatocellular carcinoma. Combination of interferon with ribavirin is the standard treatment protocol but about half patients do not mount a sustained response. Hence, unveiling the host factors that are involved in HCV infection is important to discover new strategies to prevent HCV infection and to develop new targeting treatment regimens. The non-coding microRNA (miRNA) has shown to involve in the regulation of hepatitis C virus (HCV) infection and offers an alternative target for developing anti-HCV agent. Recently, using bioinformatic analyses, several liver-abundant miRNAs with target sequences on HCV genome were selective. The anti-HCV analysis was performed by HCV subgenomic replicon cells and J6/JFH-based HCVcc virus system. One of the selective miRNA suppressed HCV replicon activity and down-regulated HCV accumulation leading to reduced infectivity of HCVcc. In addition, the miRNA could be induced by HCV infection. However, the molecular mechanisms regarding the miRNA modulates HCV infection are still not yet completely understood. In this study, we will identify the target sequence of the miRNA with an aim to unveil the miRNA regulatory mechanisms in HCV infection. At first year, the HCV genome will be subjected to mutational analysis to identify the miRNA binding sites. The association of the miRNA and HCV genome physiologically will be confirmed by Ago2-coimmunoprecipitation (Ago2-IP) analysis. At the second year, the promoter of the miRNA will be cloned and the activator which induced the promoter activity during HCV infection will be defined. At the third year, the cellular target of the miRNA which may modulate HCV infectivity will be analysis by systemic analysis by microarray and bioinformatic approach. Together, this study we hope to get a more complete picture to understand the roles of the novel cellular miRNA in anti-HCV activity and shed new insight to understanding the role of host factors in HCV pathogenesis.
Appears in Collections:	[Department of Medical Laboratory Science and Biotechnology ] Research reports

Files in This Item:

File	Description	Size	Format
index.html		0Kb	HTML	149	View/Open

Loading...