UNASSIGNED: The differential expression profiles of lncRNA in EV-D68-infected and uninfected rhabdomyosarcoma (RD) cells were studied using high-throughput sequencing technology. The knockdown through small interfering RNA (siRNA) and overexpression of lncRNA SNHG9 (small ribonucleic acid host gene 9) were applied to investigate how lncRNA SNHG9 regulates EV-D68 propagation. The targeted interactions of lncRNA SNHG9 with miR-150-5p and miR-150-5p with c-Fos were validated using dual luciferase reporter system. LncRNA SNHG9 knockdown and miR-150-5p inhibitor were co-transfected with RD cells. QRT-PCR and western blot were used to detect RNA and protein levels, of c-Fos and VP1, respectively. Median tissue culture infectious dose (TCID50) was applied to detect viral titers.
UNASSIGNED: The results demonstrated that a total of 375 lncRNAs were highly dysregulated in the EV-D68 infection model. In the EV-D68 infection model, lncRNA SNHG9 and c-Fos were increased in EV-D68-infected RD cells. However, the expression level of miR-150-5p was downregulated. In addition, overexpression of SNHG9 in RD cells resulted in decreased viral replication levels and viral titers following infection with EV-D68, and further experiments revealed that overexpression of SNHG9 inhibited the viral replication by targeting increased miR-150-5p binding and significantly increased c-Fos expression in RD cells.
UNASSIGNED: Our findings indicate that the SNHG9/miR-150-5p/c-Fos axis influences EV-D68 replication in host cells and that SNHG9 may be a possible target for anti-EV-D68 infection therapies.
UNASSIGNED:使用高通量测序技术研究了EV-D68感染和未感染的横纹肌肉瘤(RD)细胞中lncRNA的差异表达谱。通过小干扰RNA(siRNA)的敲低和lncRNASNHG9(小核糖核酸宿主基因9)的过表达用于研究lncRNASNHG9如何调节EV-D68繁殖。使用双荧光素酶报告系统验证了lncRNASNHG9与miR-150-5p和miR-150-5p与c-Fos的靶向相互作用。将LncRNASNHG9敲低和miR-150-5p抑制剂与RD细胞共转染。QRT-PCR和Westernblot检测RNA和蛋白质水平,分别为c-Fos和VP1。应用中位组织培养感染剂量(TCID50)来检测病毒滴度。
UNASSIGNED:结果表明,在EV-D68感染模型中,总共375个lncRNA高度失调。在EV-D68感染模型中,在EV-D68感染的RD细胞中,lncRNASNHG9和c-Fos增加。然而,miR-150-5p的表达水平下调。此外,在RD细胞中SNHG9的过表达导致EV-D68感染后病毒复制水平和病毒滴度降低,进一步的实验显示SNHG9的过表达通过靶向增加的miR-150-5p结合和显著增加RD细胞中的c-Fos表达来抑制病毒复制.
UNASSIGNED:我们的发现表明SNHG9/miR-150-5p/c-Fos轴影响宿主细胞中EV-D68的复制,并且SNHG9可能是抗EV-D68感染治疗的可能靶标。