摘要
凋亡蛋白(Apoptin)只诱导肿瘤细胞或转化的细胞凋亡,但其作用机制还不十分清楚.从鸡贫血病毒总DNA中克隆出凋亡蛋白基因,构建pcDNA3.1/CT-Apoptin-GFP-TOPO融合表达的载体.用该载体转染HHCC单层传代细胞24 h后,观察到绿色荧光蛋白表达,同时观察到细胞形态呈凋亡特征变化.提取细胞总RNA,按SuperArray芯片操作方法检测到实验组Bcl-2家族中的BimL,Bcl-2,Bax和Bak基因上调表达.Bcl-2家族在细胞凋亡的过程中作用重要.凋亡蛋白诱导HHCC细胞凋亡过程中,促凋亡的BimL,Bax 和Bak基因上调,抗凋亡的Bcl-2基因也上调.可能BimL与Bcl-2竞争性结合导致Bax或Bak释放,最终诱导细胞凋亡.揭示Bcl-2家族的基因参与凋亡蛋白诱导的HHCC细胞凋亡过程.
Apoptin, which only induces tumor cells or transformed cells into apoptosis, its mechanisms are unclear. The vector pcDNA3. 1/CT - GFP - apoptin - TOPO was constructed with apoptin gene that cloned from total DNA of chicken anemia virus according to its sequences in GenBank. HHCC monolayer cell lines were transfected by the vector, and GFP marker was observed by inverted fluorescence microscope at 24 h after transfection. The morphology of apoptotic cells was observed as same as special features of the apoptosis too. It implied that apoptin was expressed. Subsequently, the BimL, Bcl -2, Bax and Bak were detected by hybridization with SuperArray microarray via extracting total RNA of the cells. The genes from treatment group were up - regulated comparing with the control. The Bcl - 2 family genes are important during apoptosis. Pro - apoptotic genes, BimL, Bax and Bak were up -regulated; anti -apoptotic Bcl -2 gene was up -regulated too. Probably, BimL competitively bond with Bcl -2 and the Bax or Bak was released, and apoptosis happened; Then Bcl -2 family genes were involved in the apoptosis induced by apoptin in the HHCC cell lines.
出处
《哈尔滨工业大学学报》
EI
CAS
CSCD
北大核心
2005年第9期1180-1182,共3页
Journal of Harbin Institute of Technology
基金
黑龙江省自然科学基金资助项目(C203).
作者简介
E—mail：kuotai99@163．com；国泰（1966-），男，博士；
E-mail：kuotai99@163．com；杨谦（1959-），男，教授，博士生导师．
E-mail：kuotai99@163．com；
