中国实用医刊
中國實用醫刊
중국실용의간
CENTRAL PLAINS MEDICAL JOURNAL
2011年
17期
1-4,9
,共5页
聂常富%韩风%邱大鹏%王云检%蒙博%陈规划
聶常富%韓風%邱大鵬%王雲檢%矇博%陳規劃
섭상부%한풍%구대붕%왕운검%몽박%진규화
Real-time RT-PCR%RNA干扰%人类白细胞抗原-E
Real-time RT-PCR%RNA榦擾%人類白細胞抗原-E
Real-time RT-PCR%RNA간우%인류백세포항원-E
Real - time RT - PCR%RNA interference%Human leukocyte antigen - E
目的 应用一步法Real - time RT - PCR检测不同载体转染HepG2细胞沉默人类白细胞抗原-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、Lipofectamine 2000/psiRNA、裸psiRNA转染HepG2细胞,提取细胞总RNA,一步法Real - time RT - 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 - time RT - 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.
目的 應用一步法Real - time RT - PCR檢測不同載體轉染HepG2細胞沉默人類白細胞抗原-E (HLA -E)基因的效率.方法 分彆用Gal - PEG - PEI/psiRNA、Gal - PEG - PEI/NC - psiRNA、PEG - PEI/psiRNA、Lipofectamine 2000/psiRNA、裸psiRNA轉染HepG2細胞,提取細胞總RNA,一步法Real - time RT - PCR檢測HLA -E基因沉默前後mRNA錶達水平.結果 Gal - PEG - PEI組對HLA -E基因的沉默效率為55％,顯著高于PEG - PEI組、Lipofectamine2000組、裸psiRNA組和陰性對照組(P＜0.01).Gal-PEG-PEI組的沉默作用可持續7d.結論 經一步法Real - time RT - PCR檢測,Gal - PEG - PEI/psiRNA納米粒對HepG2細胞HLA-E基因有較高的沉默效率.
목적 응용일보법Real - time RT - PCR검측불동재체전염HepG2세포침묵인류백세포항원-E (HLA -E)기인적효솔.방법 분별용Gal - PEG - PEI/psiRNA、Gal - PEG - PEI/NC - psiRNA、PEG - PEI/psiRNA、Lipofectamine 2000/psiRNA、라psiRNA전염HepG2세포,제취세포총RNA,일보법Real - time RT - PCR검측HLA -E기인침묵전후mRNA표체수평.결과 Gal - PEG - PEI조대HLA -E기인적침묵효솔위55％,현저고우PEG - PEI조、Lipofectamine2000조、라psiRNA조화음성대조조(P＜0.01).Gal-PEG-PEI조적침묵작용가지속7d.결론 경일보법Real - time RT - PCR검측,Gal - PEG - PEI/psiRNA납미립대HepG2세포HLA-E기인유교고적침묵효솔.
Objective To evaluate interference efficiency of HLA - E mRNA expression in HepG2 cell transfected with different 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 ＜ 0.01 ),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.
