MicroRNA-98促进猪圆环病毒2型在3D4/21细胞中的复制

miRNA-98 enhances the replication of porcine circovirus type 2 in 3D4/21 cells

摘要:

[目的] 通过高通量测序的方法获得PCV2感染3D4/21细胞的miRNAs表达谱，并探讨miRNA-98在PCV2复制中的作用。[方法] 本研究以猪肺泡巨噬细胞系3D4/21细胞为细胞模型，对PCV2感染过程中的3D4/21细胞进行miRNAs差异表达分析，筛选与病毒复制相关的特异性miRNAs，并探讨其在PCV2复制中的作用。[结果] 经高通量测序，获得PCV2感染3D4/21细胞的miRNAs表达谱，结合实验室前期研究筛选获得miRNA-98。实验表明，miRNA-98的表达量随PCV2感染时间的延长而持续升高，其变化趋势与Cap蛋白表达变化基本一致，由此推测miRNA-98与PCV2复制正相关。过表达miRNA-98可显著上调Cap蛋白的表达量和PCV2的复制。进一步的研究表明，miRNA-98参与调节宿主免疫相关细胞因子的表达和PCV2的复制。[结论] miRNA-98可通过调节免疫相关细胞因子的表达调控宿主免疫功能，帮助PCV2逃逸宿主免疫，促进PCV2在3D4/21细胞中的复制。这些发现不仅为深入了解PCV2与宿主之间的关系提供了新视角，还有望为猪圆环病毒相关疾病的防控提供新的抗病毒策略。

Abstract:

[Objective] This study aims to obtain the miRNAs gene expression profiling of 3D4/21 cells infected with porcine circovirus type 2 (PCV2) and to explore the role of miRNA-98 in the replication of PCV2.[Methods] We used the porcine alveolar macrophage cell lines, 3D4/21 cells, as a cell model to analyze the different miRNAs of 3D4/21 cells infected or uninfected with PCV2. Combined with the results of bioinformatics and experiments, we screened the specific miRNAs related to virus replication and studied the role of miRNA-98 in PCV2 replication.[Results] Differential expression of miRNAs in the process of PCV2 infection was got by high-throughput sequencing technology and evaluated by qRT-RCR and preliminary study. The expression of miRNA-98 increased with the prolongation of PCV2 infection time, and the change trend of miRNA-98 was basically same as that of cap protein of PCV2. These results indicated that a significantly positive correlation between miRNA-98 expression and PCV2 replication. Overexpression of miRNA-98 significantly enhanced PCV2 replication and the expression of Cap. Further evidence supports a link between infection and the host immune regulatory cytokines, which is regulated by miRNA-98.[Conclusion] Overall, miRNA-98 can help PCV2 escape from immune surveillance by regulating the host immune system and enhance PCV2 replication. These results not only supply new insights about relationship between PCV2 infection and host but also provide a potential target for antiviral therapy against PCVAD (porcine circovirus-associated diseases).