脱氧核酶抑制鼻咽癌细胞EBV-LMP1的表达

Inhibition of EBV-LMP1 expression in NPC cells by DNAzyme

目的探讨10～23脱氧核酶(10～23DRz)对鼻咽癌生长的抑制作用。方法设计合成针对EBV-LMP1的10～23DRz,并对其进行硫代磷酸化修饰,同时针对该位点设计突变型DRz和反义寡核苷酸,经脂质体转染C666-1细胞中观察其对LMP1基因因表达的抑制效应;建立裸鼠皮下鼻咽癌移植瘤,瘤体内注射脱氧核酶及其类似物,观察其对LMP1基因因及肿瘤生长的抑制效应。结果有效转染后,脱氧核酶在细胞内抑制LMP1基因因表达;成功建立裸鼠移植瘤模型后,脱氧核酶能高效抑制LMP1基因因表达,并能抑制肿瘤生长,其作用较突变型脱氧核酶和反义寡核苷酸强(P<0.05)。结论脱氧核酶在鼻咽癌细胞中及移植瘤模型中都能高效抑制LMP1的表达,可能成为一种高度特异性的、高效的基因治疗剂。

Objective To explore the inhibitting effect of 10～23 DNAzyme on the expression of EBV-LMP1 in nasopharyngeal carcinoma（NPC） cell.Methods Phosphorothioate 10～23 DNAzyme and analogs targeting at EBV-LMP1（Epstein-Barr virus encoded latent membrane protein 1）gene mRNA were designed and then synthesized and then transfected into C666-1 cells with lipofectamine.Xenografts of NPC in nude mice were prepared,and after DNAzyme and analogs were injected into tumor,the depressive effects on the growth of tumor and LMP1 gene were assessed.Results DNAzyme inhibited the expression of LMP1 gene in vivo and in vitro,and also the growth and metastasis of xenograft of NPC were inhibited and it was found that the effect of DNAzyme was better than its analogs（P0.05）.Conclusion10～23DRz can effectively block the expression of EBV-LMP1 gene in the C666-1 cell model and xenograft in nude mice.