摘要
[目的]探究MicroRNA-1285(miR-1285)及其靶标DDX3X在猪塞内卡病毒(Senecavirus A,SVA)感染PK-15细胞中的调控作用。[方法]利用qRT-PCR、双荧光素酶活性及Western blot等方法研究miR-1285和DDX3X对I型干扰素(IFN-β)分泌及RIG-I信号通路的作用,分析miR-1285及DDX3X对SVA 3C蛋白基因表达的影响。[结果]SVA感染PK-15细胞后,miR-1285表达量显著升高,并且miR-1285与DDX3X存在负靶向关系,二者可促进IFN-β转录及蛋白水平的表达。miR-1285通过靶向DDX3X对RIG-I信号通中的MAVS、TRAF3信号分子起调控作用。对于SVA 3C蛋白基因,DDX3X可以显著抑制其转录,并且可以逆转miR-1285所诱导的上调趋势。[结论]SVA感染PK-15细胞后,宿主miR-1285及其靶标DDX3X对IFN-β及病毒3C蛋白的表达具有调控作用,研究结果将为明确miRNAs调控SVA感染的分子机制奠定基础,并为SVA的防控和诊断提供新的科学依据。
【Objective】To explore the regulation roles of microRNA-1285(miR-1285)and its target DDX3X in Senecavirus A(SVA)infected PK-15 cells.【Method】By qRT-PCR,double luciferase activity and Western blot,the effects of miR-1285 and its target DDX3X on IFN-βsecretion and the RIG-I signaling pathway were studied,and their effects on the expression of SVA 3C protein gene were analyzed.【Result】In SVA infected PK-15 cells,the expression of miR-1285 increased significantly,and there was a negative targeting relationship between miR-1285 and DDX3X.Both miR-1285 and DDX3X promoted the transcription and protein expression of IFN-β.MiR-1285 regulated MAVS and TRAF3 signaling molecules in the RIG-I signaling pathway by targeting DDX3X.For SVA 3C protein,DDX3X significantly inhibited the transcription of 3C and reversed the up-regulation trend induced by miR-1285.【Conclusion】After infecting PK-15 cells with SVA,host miR-1285 and its target DDX3X can regulate the expression of IFN-βand the viral 3C protein,which will lay a foundation for clarifying the molecular mechanism of miRNAs regulating SVA infection,and provide a new scientific basis for the prevention,control and diagnosis of SVA.
作者
孙媛
唐晓钰
白杨
陈雨琪
郑瑶瑶
吴佼玲
蓝天
马静云
SUN Yuan;TANG Xiaoyu;BAI Yang;CHEN Yuqi;ZHENG Yaoyao;WU Jiaoling;LAN Tian;MA Jingyun(College of Animal Science,South China Agricultural University,Guangzhou 510642,China)
出处
《华南农业大学学报》
CAS
CSCD
北大核心
2023年第3期357-366,共10页
Journal of South China Agricultural University
基金
广东省基础与应用基础研究基金(2020A1515010295,2022A1515012473)。
