一步法Real-timeRT-PCR检测HepG2细胞人类白细胞抗原-E基因沉默效率  

Detection of interfering efficiency of HLA - E mRNA expression in HepG2 cell using one - step real time RT - PCR method

作　　者：聂常富[1] 韩风[1] 邱大鹏[1] 王云检[1] 蒙博[1] 陈规划[2] 

机构地区：[1]河南省肿瘤医院肝胆胰外科,郑州450008 [2]中山大学附属第三医院肝移植中心

出　　处：《中国实用医刊》2011年第17期1-4,9,共5页Chinese Journal of Practical Medicine

基　　金：国家自然科学基金资助项目（30571769）;国家重点基础研究发展规划（国家973规划）资助项目（2003CB515507）;河南省基础与前沿技术研究计划项目（102300410056）

摘　　要：目的应用一步法Real-timeRT—PCR检测不同载体转染HepG2细胞沉默人类白细胞抗原一E（HLA—E）基因的效率。方法分别用Gal—PEG—PEI／psiRNA、Gal—PEG—PEI／NC—psiRNA、PEG—PEI／psiRNA、Lipofectamine2000／psiRNA、裸psiRNA转染HepG2细胞，提取细胞总RNA，一步法Real—timeRT—PCR检测HLA—E基因沉默前后mRNA表达水平。结果Gal—PEG—PEI组对HLA—E基因的沉默效率为55％，显著高于PEG—PEI组、Lipofectamine2000组、裸psiRNA组和阴性对照组（P〈0．01）。Gal—PEG～PEI组的沉默作用可持续7d。结论经一步法Real—timeRT—PCR检测，Gal—PEG—PEI／psiRNA纳米粒对HepG2细胞HLA—E基因有较高的沉默效率。Objective To HepG2 cell transfected with different evaluate interference efficiency of HLA- E mRNA expression in psiRNA carrier using one - step real time RT - PCR method, Methods HepG2 cells were transfected by Gal - PEG - PEI/psiRNA, Gal - PEG - PEI/NC - psiRNA, PEG- PEI/psiRNA, Lipofectamine 2000/psiRNA and naked -psiRNA respectively. Total mRNA of HepG2 cells was extracted. The expression levels of HLA - E mRNA were detected by one - step real time RT -PCR. Results The interference efficiency of psiRNA mediated by Gal -PEG -PEI targetting HLA - E was 55% on mRNA expression level. It was distinctively higher than those of Lipofectamine 2000, PEG - PEI and naked - psiRNA （ P 〈 O. 01 ）, and its interference effect can last for seven days. Conclusions The Gal -PEG- PEI/psiRNA nanospheres display perfect hepatocyte -targeting ability and have higher interference efficiency to HLA -E gene in HepG2 cells detected by one -step real time RT- PCR.

关 键 词：REAL-TIME RT-PCR RNA干扰 人类白细胞抗原-E 

分 类 号：R735.7[医药卫生—肿瘤]