干扰RIG-I表达对新城疫病毒诱导小鼠巨噬细胞RAW264.7产生TRAIL、IL-6的影响

Effect of interference with RIG-I expression on TRAIL and IL-6 production in Newcastle disease virus-activated mouse macrophages RAW264.7

目的:探讨干扰维甲酸诱导基因Ⅰ(RIG-I)表达对新城疫病毒(NDV)诱导小鼠巨噬细胞RAW264.7产生肿瘤坏死因子相关凋亡诱导配体(TRAIL)和白细胞介素-6(IL-6)的影响。方法:将小鼠巨噬细胞RAW264.7分为NDV组(加入NDV7793)、PBS组(仅加入PBS)和空白组(不予处理)。用Lipotectamine 3000分别将siRNA-RIGI1、siRNA-RIGI2和siRNA-NC转染入小鼠巨噬细胞,设为siRNA-RIGI1组、siRNA-RIGI2组和siRNA-NC组。用实时荧光定量PCR(RT-qPCR)法检测RIG-I、TRAIL和IL-6 mRNA相对表达量,酶联免疫吸附试验(ELISA)法检测细胞培养上清液中TRAIL和IL-6的含量。结果:与PBS组或空白组比较,NDV组RIG-I、TRAIL、IL-6 m RNA相对表达量及细胞上清液中TRAIL、IL-6的含量升高(P<0.05)。siRNA-RIGI1组、siRNA-RIGI2组RIG-I mRNA相对表达量低于siRNA-NC组和空白组(P<0.05)。与NDV组比较,NDV+siRNA-RIGI1组、NDV+siRNA-RIGI2组TRAIL、IL-6 m RNA相对表达量及细胞上清液中TRAIL和IL-6的含量降低(P<0.05)。结论:干扰RIG-I表达可抑制NDV诱导的小鼠巨噬细胞产生TRAIL、IL-6,RIG-I可能在NDV活化巨噬细胞抗肿瘤过程中发挥重要作用。

Objective:To investigate the effect of interference with retinoic acid inducible gene I(RIG-I)expression on the tumor necrosis factor-related apoptosis-inducing ligand(TRAIL)and interleukin-6(IL-6)production in Newcastle disease virus(NDV)-activated mouse macrophages RAW264.7.Methods:RAW264.7 mouse macrophages were divided into NDV group(adding NDV7793),PBS group(adding PBS only)and blank group(without treatment).The siRNA-RIGI1,siRNA-RIGI2 or siRNA-NC were transfected into mouse macrophages using Lipotectamine3000,and they were set as siRNA-RIGI1group,siRNA-RIGI2 group and siRNA-NC group.The mRNA expression of RIG-I,TRAIL and IL-6 was detected by RT-qPCR.The contents of TRAIL and IL-6 in cell supernatant were detected by ELISA.Results:Compared with blank group or PBS group,the mRNA expression of RIG-I,TRAIL and IL-6,and the contents of TRAIL and IL-6 in cell supernatant in NDV group were increased(P<0.05).The mRNA expression level of RIG-I in siRNA-RIGI1 and siRNA-RIGI2 groups was lower than that in siRNA-NC group and blank group(P<0.05).Compared with NDV group,the mRNA expression of TRAIL and IL-6 as well as the contents of TRAIL and IL-6 in cell supernatant in siRNA-RIGI1 and siRNA-RIGI2groups were decreased(P<0.05).Conclusion:Interference of RIG-I can inhibit the production of TRAIL and IL-6 in NDV-infected mouse macrophages,and RIG-I may play an important role in the anti-tumor process of macrophages activated by NDV.