Ring-Box 1基因负调控抗病毒天然免疫的初步研究

摘要

Objective To study the role of Ring-Box 1 (RBX1/ROC1) in regulating retinoic acid-inducible gene Ⅰ (DDX58/RIG-Ⅰ) mediated anti-viral signaling pathway and explore the potential therapeutic target.Methods The full-length RBX1/ROC1 gene was amplified by PCR and inserted it in pCDNA3 plasmid. And then, the over-expression of RBX1/ROC1 in HEK293 cells was detected by Western Blot. Finally, the regulatory role of RBX1/ROC1 on DDX58/RIG-Ⅰ was detected by luciferase reporter assays.Results Flag-ROC1 eukaryotic expression plasmid was constracted. After transfected it into the HEK293 cells, Flag-ROC1 was expressed in these cells. We further found that RBX1/ROC1 downregulated DDX58/RIG-Ⅰ signaling pathway by luciferase reporter assays.Conclusions RBX1/ROC1 is a negative regulator of DDX58/RIG-Ⅰ anti-viral signaling pathway and our study provides a potential therapeutic target of anti-viral.%目的研究Ring-Box 1基因(RBX1/ROC1)对视黄酸诱导基因-Ⅰ(retinoic acid inducible geneⅠ,DDX58/RIG-Ⅰ)抗病毒感染信号通路功能的影响,寻找抗病毒治疗的潜在靶标.方法利用PCR扩增RBX1/ROC1基因,将其插入pCDNA3载体,并且通过免疫印迹检测质粒在细胞中的表达;利用荧光素酶报告基因检测RBX1/ROC1基因对DDX58/RIG-Ⅰ信号通路的影响.结果本研究成功构建Flag-RBXI/ROC1真核表达载体并在HEK293细胞中得到了表达;通过荧光素酶报告基因分析发现RBX1/ROC1基因抑制DDX58/RIG-Ⅰ信号通路.结论RBX1/ROC1是宿主天然免疫DDX58/RIG-Ⅰ抗病毒感染信号通路的负调控分子,为寻找抗病毒靶点打下基础.