MicroRNA-21-3p modulates FGF2 to facilitate influenza A virus H5N1 replication by refraining type I interferon response
MicroRNA-21-3p modulates FGF2 to facilitate influenza A virus H5N1 replication by refraining type I interferon response
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DOI：
10.1042/bsr20200158
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发表时间：
2020-05-28
影响因子：
4
通讯作者：
Gu, Tingting
中科院分区：
文献类型：
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作者：
Shi, Jianli;Feng, Ping;Gu, Tingting
背景：近几十年来，甲型流感病毒(IAV)极大地影响了公众健康。越来越多的数据表明，宿主microRNAs(MiRNAs)与IAV的复制有关。本研究主要研究microRNA-21-3p(miR-21-3p)对H5N1病毒复制的影响。方法：采用实时定量聚合酶链式反应(qRT-PCR)检测miR-21-3p、病毒结构因子(基质1、核蛋白(NP))、I型干扰素(干扰素-β、干扰素-ETA)、干扰素刺激基因(蛋白激酶R(PKR)、粘病毒抗性A(MxA)、2‘-5’-寡聚腺苷合成酶2(OAS)、成纤维细胞生长因子2(GF2))的表达水平。Western印迹法检测M1、NP、FGF2蛋白水平。用组织培养感染剂量50%(TCID50)法检测病毒滴度。采用双荧光素酶报告实验和核糖核酸免疫沉淀(RIP)实验验证miR-21-3p与FGF2的相互作用。结果：MIR-21-3p在H5N1感染者和A549细胞中表达下调。MIR-21-3p过表达促进了H5N1感染A549细胞M1、NP、TCID50的水平，降低了干扰素-β、干扰素-α、PKR、Mxa和OAS的水平。FGF2被证实是miR-21-3p的直接靶点。FGF2的引入抵消了miR-21-3p介导的H5N1感染A549细胞M1、NP和TCID50水平的下降，以及干扰素-β、干扰素-α、PKR、Mxa和OAS水平的下降。结论：MIR-21-3p下调FGF2的表达，促进H5N1复制，抑制干扰素应答。
Background: Influenza A virus (IAV) has greatly affected public health in recent decades. Accumulating data indicated that host microRNAs (miRNAs) were related to IAV replication. The present study mainly focused on the effects of microRNA-21-3p (miR-21-3p) on H5N1 replication.Methods: The levels of miR-21-3p, virus structural factors (matrix 1 (M1), nucleoprotein (NP)), type I interferon (IFN) response markers (IFN-beta, IFN-eta), IFN-stimulated genes (protein kinase R (PKR), myxovirus resistance A (MxA), 2 '-5 '-oligoadenylate synthetase 2 (OAS)), and fibroblast growth factor 2 (FGF2) were measured by quantitative real-time polymerase chain reaction (qRT-PCR). The protein levels of M1, NP, and FGF2 were tested by Western blot assay. The virus titer was assessed by tissue culture infective dose 50% (TCID50) assay. The dual-luciferase reporter assay and ribonucleic acid (RNA) immunoprecipitation (RIP) assay were used to verify the interaction between miR-21-3p and FGF2.Results: MiR-21-3p was reduced in H5N1-infected patients and A549 cells. MiR-21-3p overexpression facilitated the levels of M1, NP, TCID50 value, and reduced the levels of IFN-beta, IFN-alpha, PKR, MxA, and OAS in H5N1-infected A549 cells. FGF2 was verified as a direct target of miR-21-3p. The introduction of FGF2 counteracted miR-21-3p-mediated decrease in the levels of M1, NP, and TCID50 value, as well as reduction in the levels of IFN-beta, IFN-alpha, PKR, MxA, and OAS in H5N1-infected A549 cells.Conclusion: MiR-21-3p down-regulated FGF2 expression to accelerate H5N1 replication and confine IFN response.