MicroRNA-1285及其靶标DDX3X对猪塞内卡病毒感染PK-15细胞的调控作用

    Regulation effects of microRNA-1285 and its target DDX3X on Senecavirus A infected PK-15 cells

    • 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。

       

      Abstract:
      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-β secretion 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-β and 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.

       

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