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| 细胞浆中PrP蛋白的表达及其细胞毒性作用的研究 | |
| 引用本文： | 王新,董辰方,石琦,石崧,王桂荣,雷艳君,安润,徐琨,姜慧英,韩俊,赵玉军,董小平.细胞浆中PrP蛋白的表达及其细胞毒性作用的研究[J].病毒学报,2008,24(4):277-281. |
| 作者姓名： | 王新 董辰方 石琦 石崧 王桂荣 雷艳君 安润 徐琨 姜慧英 韩俊 赵玉军 董小平 |
| 作者单位： | 1. 沈阳农业大学,畜牧兽医学院,沈阳,110161;中国疾病预防控制中心,病毒病预防控制所,传染病预防控制国家重点实验室,北京,100052 2. 中国疾病预防控制中心,病毒病预防控制所,传染病预防控制国家重点实验室,北京,100052 3. 中国疾病预防控制中心,病毒病预防控制所,传染病预防控制国家重点实验室,北京,100052;西安交通大学,医学院,西安,710061 4. 沈阳农业大学,畜牧兽医学院,沈阳,110161 |
| 基金项目： | 国家科技攻关计划，国家重点基础研究发展计划(973计划)，国家自然科学基金 |
| 摘 要： | 为了探讨在胞浆中表达的PrP的理化特征以及对细胞活性的影响,我们构建了胞浆型PrP(CytoPrP)真核表达质粒,瞬时转染人神经母细胞瘤SH-SY5Y细胞,通过蛋白酶敏感性实验检测CytoPrP及其蛋白酶抗性,利用MTT和Trypan Blue细胞计数检测CytoPrP的细胞毒性作用.结果显示,CytoPrP在胞浆内的存在受蛋白酶抑制剂的控制;与野生型PrP相比,CytoPrP具有相对较强的蛋白酶K(PK)抗性.MTT和细胞计数实验均显示,Cyto-PrP的存在可诱导明显的细胞毒性效应,而CytoPrP的细胞毒性作用受蛋白酶抑制剂含量的影响,呈剂量依赖关系.上述结果为研究CytoPrP在朊病毒病的发病机制中的意义提供了一定的科学数据. |
| 关 键 词： | 朊病毒 胞浆PrP 细胞毒作用 蛋白酶抗性 |
Expression of Cytosolic PrP and Analysis of Its Cytotoxic Activities |
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| WANG Xin,DONG Chen-fang,SHI Qi,SHI Song,WANG Gui-rong,LEI Yan-jun,AN Run,XU Kun,JIANG Hui-ying,HAN Jun,ZHAO Yun-jun,DONG Xiao-ping.Expression of Cytosolic PrP and Analysis of Its Cytotoxic Activities[J].Chinese Journal of Virology,2008,24(4):277-281. | |
| Authors: | WANG Xin DONG Chen-fang SHI Qi SHI Song WANG Gui-rong LEI Yan-jun AN Run XU Kun JIANG Hui-ying HAN Jun ZHAO Yun-jun DONG Xiao-ping |
| Institution: | College of Animal Husbandry and Veterinary Medicine, Shenyang Agricultural University, Shenyang 110161, China. wangxin82-82@163.com |
| Abstract: | In order to study the physicochemical characteristics of cytosolic PrP(CytoPrP) and evaluate its possible influence on cell viability,a recombinant plasmid expressing human CytoPrP eukaryoticly was constructed and transfected into human neuroblastoma cell line SH-SY5Y transiently.Proteinase-resistant activities of CytoPrP were evaluated by a proteinase K(PK) digestion and cytotoxic effects of CytoPrP were tested by MTT assay and Trypan Blue cell-counting.The presence of CytoPrP in cytoplasm after transfection was controlled by the presence of protease inhibitor.Compared with wild-type PrP,CytoPrP possessed relatively stronger PK-resistant activities.Obvious cytotoxic effects were observed in the cells after inducement of CytoPrP in cytoplasm by protease inhibitor,showing a dose-dependent manner.The results provide useful scientific evidences for further studies of potential role of CytoPrP in pathological mechanism of prion disease. |
| Keywords: | Prion cytosolic PrP cytotoxic effect proteinase-resistant characteristics |
| 本文献已被 CNKI 维普 万方数据 等数据库收录！ | |