| shRNA干扰肝癌细胞Kir6.2基因表达载体的构建及鉴定 |
| |
| 引用本文： | 苏晓通,梁平,李靖,丁生才,杨彤翰. shRNA干扰肝癌细胞Kir6.2基因表达载体的构建及鉴定[J]. 西南国防医药, 2008, 18(3): 340-343 |
| |
| 作者姓名： | 苏晓通 梁平 李靖 丁生才 杨彤翰 |
| |
| 作者单位： | 第三军医大学新桥医院肝胆外科,重庆,400037 |
| |
| 摘 要： | 目的构建shRNA体内表达载体pGenesil-3-siRNA,筛选出抑制Kir6.2基因表达的有效靶序列.方法以Kir6.2基因为目的基因,以产生shRNA质粒载体pGenesil-3为表达模板,细胞内转录合成2条siRNA,脂质体转染法将shRNA质粒载体pGenesil-3共转染SK-Hep1细胞,将细胞分为SK组(未转染)、HK组(转染阴性对照质粒)、K1组(转染重组质粒K1)和K2(转染重组质粒K2)组.经G418筛选出单克隆进行培养,用RT-PCR和Western Blotting分别检测Kir6.2 mRNA和蛋白的表达,筛选出抑制Kir6.2表达的有效siRNA靶序列.结果SK、HK、K1和K2组Kir6.2 mRNA表达的相对量分别为0.936±0.042、0.848±0.058、0.225±0.0360、0.171±0.029,K1、K2组Kir6.2 mRNA的表达与HK、SK组比较均明显降低(P＜0.05).SK、HK、K1和K2组的Kir6.2蛋白表达的量分别为0.925±0.045、0.901±0.067、0.269±0.024、0.318±0.031,K1、K2组Kir6.2蛋白的量与HK、SK组比较均明显降低(P＜0.05).结论shRNA抑制了肝癌细胞Kir6.2基因的表达.
|
| 关 键 词： | RNA干扰 肝细胞癌 RT-PCR Western Blotting |
| 文章编号： | 1004-0188(2008)03-0340-04 |
| 修稿时间： | 2008-02-01 |
Construction and identification of vector against Kir6.2 gene expression in hepatoma cells by short interfering RNAs |
| |
| SU Xiao-tong,LIANG Ping,LI Jing,DING Sheng-cai,YANG Tong-han. Construction and identification of vector against Kir6.2 gene expression in hepatoma cells by short interfering RNAs[J]. Medical Journal of National Defending forces in Southwest China, 2008, 18(3): 340-343 |
| |
| Authors: | SU Xiao-tong LIANG Ping LI Jing DING Sheng-cai YANG Tong-han |
| |
| Affiliation: | SU Xiao-tong,LIANG Ping,LI Jing,DING Sheng-cai,YANG Tong-han Department of Hepatobiliary Surgery,Xinqiao Hospital,Third Military Medical University,Chongqing,400037,China. |
| |
| Abstract: | |
| |
| Keywords: | RT-PCR Western Blotting |
| 本文献已被 CNKI 万方数据 等数据库收录！ |
|
