人PrP基因体外诱导细胞凋亡的初步研究  

Expression of Human PrP Gene in CHO Induces the Features of Apoptosis

作　　者：赵霞[1] 董小平[1] 周伟[1] 洪涛[1] 

机构地区：[1]中国预防医学科学院病毒学研究所,北京100052

出　　处：《病毒学报》2000年第3期207-211,共5页Chinese Journal of Virology

基　　金：国家863高技术研究发展计划!( 863 -10 2 -110 -0 0 3-0 5 )

摘　　要：朊蛋白为可传播性海绵样脑病的感染因子。将编码朊病毒蛋白的PrP标准及突变DNA序列分别连接到真核表达载体 pcDNA3 1中 ,并分别转入中国仓鼠卵巢细胞 (CHO)。经过RT -PCR、RNAdotblot、Westernblot鉴定证实 ,得到了稳定表达人标准 (CHOs)和终止密码突变PrP基因 (CHOm)的细胞系。对此细胞系进一步研究发现 ,带有标准人PrP序列的CHO细胞系的生长速度比其它对照细胞系为慢 ,但细胞周期无明显变化。用全反式维甲酸处理细胞后 ,CHOs细胞系呈现出Hoechest332 5 8核质深染、DNAladder电泳图形及细胞流式光度术分析凋亡细胞群和G1细胞亚群增加 ,而在同样的处理条件下对照细胞系未出现凋亡征象。这提示朊蛋白可能参与细胞程序化死亡的调控过程。Prion is considered to be the infectious agent for the transmissible spongiform encephalopathy. The standard and mutated human PrP genes were separately inserted into an eukaryotic expression vector and transfected into CHO cell, respectively, generating CHOs strain containing the standard PrP gene and CHOm strain containing the mutated PrP gene. Analysis of the cellular DNA, RNA and protein revealed that the transfected human PrP gene was persistently transcribed and expressed in CHOs strain. During the passages, the CHOs cell line showed a slowly duplicating rate as compared with the mock, but the CHOc containing only expression vector and the CHOm showed not any changes of cell life cycles. After treatment with RA, the cell line CHOs showed deeply stained chromatin with Hoechst 33258. DNA ladder pattern in agarose gel electrophoresis and the flow cytometry both revealed clusters of apoptotic cells and increasing amount of G1 subgroup cells, whereas the other CHO cell lines failed to show any features of apoptosis. It suggests that human prion protein might play a role in controlling cell programmed death.

关 键 词：朊蛋白 基因转染 细胞凋亡 人PrP基因 朊病毒病 

分 类 号：S852.659.7[农业科学—基础兽医学] R373[农业科学—兽医学]