小发卡状RNA联合脱氧核酶抑制鼻咽癌细胞中EB病毒潜伏膜蛋白基因表达研究

The effect of short hairpin RNA and DNAzyme gene therapy on latent membrane protein-1 expression in nasopharyngeal carcinoma

目的:探讨小发卡状RNA单独及联合脱氧核酶对鼻咽癌细胞中EB病毒EBV潜伏膜蛋白1(LMP1)基因表达的抑制作用。方法:带绿色荧光报告基团的LMP1mRNA表达载体,小发卡状RNA及脱氧核酶按照阳性对照组、RNA干扰组、联合组和脱氧核酶组4组分别共转染HNE1细胞,观察分析计数细胞中绿色荧光表达情况,检测LMP1mRNA和蛋白表达水平。结果:联合组细胞绿色荧光OD值均数与RNA干扰组差异有统计学意义(P<0.01);联合组绿色荧光抑制率分别为86.31%和88.88%,比RNA干扰组抑制率75.15%高;EBVLMP1mRNA和蛋白水平检测提示联合组抑制效率较RNA干扰组高。结论:小发卡状RNA联合脱氧核酶能够提高抑制EBV-LMP1基因表达的效率。

Objective：To study the inhibitory effects of short hairpin RNA（pshLMP1） combined with DNAzyme or alone on latent membrane protein-1 （LMP1）expression in HNE1 cell lines.Method：The pshLMP1 and DNAzymes were co-transfected with pEGFP-N1-1158 （a expression vector encoding LMP1 and EGFP fusion protein） into HNE1 cells,divided into 4 groups as positive control,RNA interference,combining and DNAzyme groups.The green fluorescence were analyzed and then the mRNA and protein of LMP1 gene were detected.Result：The HNE1 cells expressing green fluorescence in combining group were significantly less than that in RNA interference group（P0.01）;The inhibition efficiency of green fluorescence in combining group were 86.31%和88.88%,which were higher than that in RNA interference group with 75.15%.The detection of LMP1 mRNA and proteins showed that combining group had a higher inhibition ability.Conclusion：Combining gene therapy with pshLMP1 and DNAzyme is superior to pshLMP1 alone to inhibit LMP1 gene expression.