| 介导穿孔素短发夹RNA重组腺病毒的构建及其生物学作用 |
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| 引用本文： | 赵莉琳,刘阳. 介导穿孔素短发夹RNA重组腺病毒的构建及其生物学作用[J]. 细胞与分子免疫学杂志, 2010, 26(3) |
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| 作者姓名： | 赵莉琳 刘阳 |
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| 作者单位： | 1. 重庆医科大学,附属第一医院口腔科,重庆,400016
2. 重庆医科大学,附属第二医院耳鼻咽喉-头颈外科,重庆,400010 |
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| 摘 要： | 目的:采用腺病毒介导的RNAi技术对穿孔素(PF)的功能进行体外研究,以期为深入研究PF的作用机制奠定基础.方法:构建腺病毒介导的穿孔素shRNA(Ad-shPF).首先通过mRNA水平验证所设计的pShuttle-shPF序列的有效性,有效后包装Ad-shPF,验证其对NK-92细胞穿孔素mRNA和蛋白表达水平的影响.结果:转染pShuttle-shPF(1-3)的干扰效率分别为(74.2±4.1)%、(43.2±3.2)%、(61.7±2.6)%.干扰效果明显,其中又以pShuttle-shPF1的干扰效果最佳.成功包装Ad-shPF1,感染NK-92细胞,mRNA表达水平降至对照组的32%;Western blot分析表明Ad-shPF1处理后可在蛋白水平上明显抑制PF的表达.结论:PF shRNA重组腺病毒可以在mRNA和蛋白水平抑制NK-92细胞中PF的表达.
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| 关 键 词： | 穿孔素 腺病毒NK-92细胞 短发夹RNA |
Recombinant Adenovirus mediate RNA silencing of perforin in NK-92 cells and its function |
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| ZHAO Li-lin,LIU Yang. Recombinant Adenovirus mediate RNA silencing of perforin in NK-92 cells and its function[J]. Chinese journal of cellular and molecular immunology, 2010, 26(3) |
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| Authors: | ZHAO Li-lin LIU Yang |
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| Affiliation: | ZHAO Li-lin1,LIU Yang21First Affiliated Hospital,Chongqing Medical University,Chongqing 400016,2Department of Otolaryngology-Head , Neck Surgery,Second Affiliated Hospital,Chongqing 400015,China |
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| Abstract: | AIM:To adenovirus-mediated RNAi technology be used to down-regulate perforin expression in NK-92 cells.This study will provide convenience for further investigation of the mechanism of perforin.METHODS:First,we designed three perforin shRNAs(shPF) and identified their knockdown efficiency at mRNA levels.Then we constructed recombinant adenovirus with shPF.After that,we transfected NK-92 cells with Ad-shPF and detected the expression of perforin at mRNA and protein levels.RESULTS:We confirm that the design o... |
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| Keywords: | perforin adonovirus NK-92 cell short hairpin RNA |
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