miRNA-155在ALV-J与REV共感染中对肌动蛋白细胞骨架通路的影响

Effects of miRNA-155 on the Cytoskeletal Pathways in REV and ALV-J Co-Infection

该研究探讨了miRNA-155的表达在J亚群禽白血病病毒(subgroup J avian leucosis virus, ALV-J)和网状内皮组织增生症病毒(reticuloendotheliosis virus, REV)共感染中对肌动蛋白细胞骨架通路的影响。通过蛋白质组学和转录组学检测技术对ALV-J、REV和ALV-J+REV感染DF-1细胞分泌的外泌体进行蛋白质和miRNA定量差异表达综合分析,筛选共感染组与单独感染组相比共同变化的因子及其参与的信号通路。结果显示, ALV-J和REV的共感染对彼此具有协同效应, REV对ALV-J的协同起主导作用; GTP结合蛋白J(GTP-binding protein J, RhoJ)、NCK相关蛋白1(NCKassociation protein 1, NCKAP1)、肌动蛋白相关2/3复合体亚基5(actin-related 2/3 complex subunit5, ARPC5)和miRNA-155的靶蛋白整合蛋白(integrin, ITG)共同参与肌动蛋白细胞骨架通路。体外转染促进或抑制miRNA-155表达,采用RT-PCR检测其对病毒复制以及ITG、RhoJ、NCKAP1和ARPC5表达的影响。结果显示,促进miRNA-155的表达时,有利于病毒的复制, RhoJ和NCKAP1的表达上升,而ITG和ARPC5表达下降;抑制mi RNA-155的表达时,则抑制病毒复制, RhoJ和NCKAP1的表达下降, ITG和ARPC5表达上升;且共感染组中变化更为显著,这与前期病毒感染引起的相关蛋白变化结果相吻合。该研究结果表明, ALV-J与REV共感染时的协同促进作用可能与miRNA-155调节肌动蛋白细胞骨架通路中4种蛋白的表达变化密切相关, miRNA-155是两种病毒协同作用的关键调节因子。

This study was aimed to investigate the effects of miRNA-155 expression on actin cytoskeletal pathways in ALV-J and REV co-infection. The exosomes from ALV-J, REV and ALV-J＋REV infected DF-1 cells were used to quantity the differentially expressed proteins and miRNAs by proteomic and transcriptome detection technology. Then, the common changed factor and their participation in the signal pathway in co-infected group compared with single infected group were screened. The results showed that ALV-J and REV had synergistic effects on each other, and REV played a leading role in synergism on ALV-J. RhoJ, NACKP1 and ARPC5, and the miRNA-155 target protein ITG were involved in the actin cytoskeleton pathway. After transfection of miRNA-155 inhibitors or mimics, RT-PCR was used to detect the effects of miRNA-155 expression on the virus replication, and the expression of ITG, RhoJ, NCKAP 1 and ARPC5 as well. The results demonstrated that the virus replication and the expression of RhoJ and NCKAP1 were promoted, whereas, ITG and ARPC5 expression was decreased, when the miRNA- 155 expression was increased. Inhibition of miRNA-155 expression inhibited virus replication, blocked the expression of RhoJ and NCKAP1, and increased the expression of ITG and ARPC5. These changes were more significant in the co-infected group, which was consistent with the previous results that the changes of associated proteins induced by virus infection. Our results suggested that the synergistic effect of ALV-J and REV co-infection might be associated with the miRNA-155 regulation the 4 proteins in actin cytoskeleton pathway. The miRNA-155 is a key factor regulating the synergistic effects of the two viruses.