Efficient delivery of siRNA into cytokine-stimulated insulinoma cells silences Fas expression and inhibits Fas-mediated apoptosis
Efficient delivery of siRNA into cytokine-stimulated insulinoma cells silences Fas expression and inhibits Fas-mediated apoptosis
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DOI：
10.1016/j.febslet.2005.12.068
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发表时间：
2006-01-23
期刊：
影响因子：
3.5
通讯作者：
Zhang, YC
中科院分区：
文献类型：
--
作者：
Burkhardt, BR;Lyle, R;Zhang, YC
Fas/FasL相互作用被认为是I型糖尿病中P细胞死亡的重要机制。最近的研究表明，由小干扰RNA（siRNA）提供的RNA干扰可以在哺乳动物细胞中提供特异性和稳健的基因沉默。本研究旨在探讨siRNA沉默Fas表达对细胞因子孵育后Fas介导的小鼠胰岛素瘤细胞凋亡的影响。我们的结果表明，siRNA能够快速抑制苦参碱诱导的Fas mRNA的产生和细胞表面Fas蛋白。仅在与siRNA长时间孵育后观察到总Fas蛋白的完全抑制，表明Fas蛋白的缓慢翻转。此外，siRNA显着抑制Fas介导的β细胞凋亡的Caspase-3和末端脱氧核苷酸转移酶生物素-dUTP缺口末端标记法评估，其程度呈正相关的细胞表面Fas的水平。这些观察结果提供了额外的证据支持Fas介导的途径在β细胞破坏中的作用，并表明靶向Fas的siRNA可能在预防I型糖尿病和改善移植中的胰岛细胞活力方面具有治疗价值。(c)2005年欧洲生物化学学会联合会。Elsevier B. V.出版，保留所有权利。
Fas/FasL interactions have been proposed as a potentially important mechanism mediating P-cell death in type I diabetes. Recent investigations suggest RNA interference, afforded by small interfering RNAs (siRNA), can provide specific and robust gene silencing in mammalian cells. The current study attempted to investigate the effects of silencing Fas expression with siRNA on Fas-mediated apoptosis in mouse insulinoma cells following cytokine incubation. Our results indicate that siRNA is capable of rapid inhibition of cytokine-induced Fas mRNA production and cell surface Fas protein. A complete suppression of the total Fas protein was only observed after prolonged incubation with siRNA, suggesting a slow turn-over of Fas protein. Moreover, siRNA significantly inhibited Fas-mediated beta-cell apoptosis assessed by Caspase-3 and terminal deoxynucleotidyl transferase biotin-dUTP nick end labeling assays, the extent of which positively correlated with the level of cell surface Fas. These observations provide additional evidence supporting a role for the Fas-mediated pathway in beta-cell destruction, and suggest that siRNA targeting Fas may be of therapeutic value in preventing type I diabetes and improving islet cell viability in transplantation. (c) 2005 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.