细小病毒H-1非结构蛋白NS1在人肝癌细胞内的超显微结构定位  

Ultrastructural localization of parvovirus H-1 nonstructural protein 1 in human hepatoma cells

作　　者：施志仪[1] 林万敏[1] 李健宏[1] 罗祖玉[1] 

机构地区：[1]复旦大学生命科学学院生理学与生物物理系,上海市200433

出　　处：《世界华人消化杂志》2001年第5期529-532,共4页World Chinese Journal of Digestology

基　　金：国家自然科学基金;No.39470773

摘　　要：目的研究细小病毒H-1非结构蛋白NS1对体内人肝癌细胞的抑制作用机制。方法利用免疫胶体金标记电镜技术,在感染细小病毒H-1后不同时间,观察了病毒非结构蛋白NSl在人肝癌裸小鼠QGY-9204移植瘤模型细胞内的超显微结构定位。结果研究表明,在感染后4h,H-1病毒的NSI蛋白主要集中在核仁内;感染后12h,NSl蛋白由核仁内向常染色区转移;感染后24h,常染色区的NSl蛋白逐渐增多并且向核外扩散;感染后72h,核内和胞质内NSl蛋白越来越多,与此同时,细胞核圆缩,核仁消失.H-1 NSl蛋白表达在细胞内的定位过程与H-1DNA复制的定位相一致。用原位杂交研究表明,在感染24h,阳性杂交颗粒仅在少数细胞核内出现,在感染后72h,核内阳性杂交颗粒增多并向核外扩散。结论 NSl从核仁扩散到细胞质,辅证了NS-1蛋白在病毒生活周期和抑制肿瘤细胞生长中起重要作用。AIM To study the inhibitory effect of parvovirus H1 nonstructural protein 1 on the human hepatoma in vivo and its mechanism. METHODS The human hepatoma model QGY-9204 in nude mice was infected by autonomous parvovirus H-1. Using the immuno-colloid gold labeling technique, the ultrastructural localization of the non-structural protein 1 (NS1) was investigated at the different time after infection. RESULTS Time sequence and localization of H-1 NS1 protein was as follows: granules with colloid labelling distributed mainly in nucleolus at 4 hours post infection (4h.p.i), and started moving from there to euchromosome at 12h . p.i.; these granules increased and moved out of nucleus at 24h . p.i.; more granules appeared in nucleus and cytoplasm at 72h. p.i.. Electron micrographs showed that nucleus shrank and became round, and nucleolus degenerated. This process is coincides with the localization of H-1 DNA replication. From in situ hybridization study, the labeled particles representing H-1 DNA appeared in a few nucleus at 24h .p.i.; these particles increased and moved to cytoplasm at 72h. p.i.. CONCLUSION The dynamic distribution of H-1 NS1 protein in H-1 infected human hepatoma cells is a progress from nucleolus to total nucleus and to cytoplasm, it supported the view point that both H-1 NS-1 protein and DNA replication play important roles in the viral life cycle and inhibition of growth of hepatoma cells.

关 键 词：肝肿瘤 细小病毒H-1 非结构蛋白NS1 超显微结构定位 

分 类 号：R735.7[医药卫生—肿瘤]