郑州大学学报（医学版）2013，Vol.48Issue(2)：163-167,5.DOI:10.3969/j.issn.1671-6825.2013.02.005
Cyclin E RNA干扰载体的构建及其对EC9706细胞Cyclin E表达的影响
Construction of RNAi vector of Cyclin E and its effection on Cyclin E expression of EC9706 cell
王娜 1冷弘 1李敏 1臧文巧 1王媛媛1
作者信息
摘要
Abstract
Aim:To construct the RNA interference vector of Cyclin E and transfect into EC9706 cell lines, and observe the supression effects on the expression of Cyclin E in transfected cell. Methods:Two pairs of small interfering RNA targeting to Cyclin E were designed. The annealed products ( dsl, ds2 ) were ligated into RNA interference vector. The recombi-nant vectors were transfected into EC9706. GFP expression of transfected cells was detected by fluorescent microscopy. mR-NA and protein expression of Cyclin E were measured by RT-PCR and Western-blot assay. Results:The 2 inserted sequences which combined into siRNA vector were the same with the target sequences designed previously. Green fluorescence of the transfected cells could be observed under fluorescent microscopy in interfered group 1, interfered group 2 and non-siR-NA associated group but no green fluorescence in blank control group. There were significant differences in the expression of Cyclin E mRNA among the four groups( F = 15.381 ,P =0.002 ). And the expression of mRNA in the interfered group 1 and
2 were significantly decreased compared with those in non-siRNA associated group and blank control group ( P <0.05 ). Con-Clusion:The recombinant vector could effectively suppress the expression of Cyclin E gene in RNA level.关键词
Cyclin E/RNA干扰/EC9706细胞Key words
Cyclin E/RNA interference/EC9706 cell分类
医药卫生引用本文复制引用
王娜,冷弘,李敏,臧文巧,王媛媛..Cyclin E RNA干扰载体的构建及其对EC9706细胞Cyclin E表达的影响[J].郑州大学学报（医学版）,2013,48(2):163-167,5.
