BimS真核质粒过表达载体构建及诱导ACC-2细胞凋亡作用  

The construction of BimS eukaryotic plasmid expression vector and its role in inducing the apoptosis of ACC-2 cells

作　　者：李旭奎[1] 姚佳[1] 饶国洲[1] 

机构地区：[1]西安交通大学口腔医院口腔颌面外科,陕西省牙颌疾病临床研究中心,西安710004

出　　处：《北京口腔医学》2016年第2期75-78,共4页Beijing Journal of Stomatology

基　　金：陕西省社发攻关资助项目(2013K12-03-21)

摘　　要：目的构建BimS真核质粒过表达载体及鉴定并转染ACC-2细胞,观察其在细胞中上调BimS蛋白表达及促凋亡作用。方法根据人BimS基因设计引物,扩增目的基因片段,挑选阳性克隆,分别采用PCR扩增电泳和DNA测序鉴定。分为对照组和实验组将构建的质粒表达载体转染ACC-2细胞,通过定量PCR和Western biot检测其基因和蛋白相对表达水平,检测细胞凋亡率和超微结构的变化。结果实验组细胞凋亡明显,凋亡率为(29.6±0.3)%;对照组细胞凋亡率仅为(0.83±0.2)%,实验组与对照组细胞凋亡率比较具有统计学差异(P<0.05)。电镜下实验组细胞呈典型的细胞凋亡形态学改变,对照组细胞形态未发生变化。结论 BimS真核质粒过表达载体构建成功,并能显著上调ACC-2细胞中BimS mRNA、蛋白表达及诱导细胞凋亡。Objective To construct and identify BimS eukaryotic plasmid expression vector,and to investigate its role in inducing apoptosis of ACC-2 cells. Methods The primers were designed according to BimS gene and gene fragment was amplifted. The positive clones were picked by PCR electrophoresis and sequencing DNA respectively. ACC-2 cells were transfected by constructed plasmid expression vector. The gene and protein expression level were examined by quantitative PCR and Western biot. The apoptosis rate and the ultrastructure were detected by flow cytometry and transmission electron microscopy. Results The apoptosis rate of the experimental group（ 29. 6 ± 0. 3） % was significantly higher than that of the control group（ 0. 83 ± 0. 2） %（ P 〈0. 05）. The experimental group showed typical apoptotic cell morphology under electron microscope,but the control group did not exhibit cell morphology change. Conclusion BimS eukaryotic expression vector plasmid was successfully constructed and could up-regulate BimS mRNA and protein expression and induce apoptosis of ACC-2 cells.

关 键 词：BimS ACC-2细胞 细胞凋亡 

分 类 号：R739.8[医药卫生—肿瘤]