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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。
AIM: To investigate the inhibitory effect of small hairpin RNA (RNA) alone and in combination with deoxyribozyme on EBV latent membrane protein 1 (LMP1) gene expression in nasopharyngeal carcinoma cells. Methods: The LMP1 mRNA expression vector with green fluorescent reporter group, hairpin RNA and DNAzyme were co-transfected into HNE1 cells according to the positive control group, RNA interference group, combination group and DNAzyme group, respectively. The number of counting cells In green fluorescent expression, detection of LMP1 mRNA and protein expression levels. Results: The mean fluorescence intensity of green fluorescence in combination group was significantly lower than that in RNA interference group (P <0.01). The inhibition rates of green fluorescence in combined group were 86.31% and 88.88%, respectively, which were 75.15% higher than those in RNA interference group. The detection of EBVLMP1mRNA and protein levels suggested that the combined group had higher inhibition efficiency than RNA interference group. Conclusion: Small hairpin RNA combined with DNAzyme can increase the efficiency of inhibiting EBV-LMP1 gene expression.