Department of the Laboratory Medicine, The Second Hospital, Wenzhou Medical University, Wenzhou, Zhejiang, China; Jinhua Municipal Central Hospital, Jinhua, Zhejiang, China.
Department of the Laboratory Medicine, The Second Hospital, Wenzhou Medical University, Wenzhou, Zhejiang, China.
Gene. 2014 Feb 25;536(2):272-8. doi: 10.1016/j.gene.2013.12.012. Epub 2013 Dec 18.
MicroRNAs (miRNAs) play important roles in regulating gene expression of plants, animals and viruses. Comprehensive characterization of host and viral miRNA will help uncover the molecular mechanisms that underlie the progression of human cytomegalovirus (HCMV) latent infection. To investigate the miRNA expression profile of HCMV and host cells during latent infection, we performed deep-sequencing analysis of the small RNAs isolated from HCMV-infected and mock-infected human monocytic leukemia cell line, THP-1.
We established a HCMV latent infection cell model using the THP-1 cells. High-throughput sequencing technology was used to sequence small RNA libraries of the HCMV-infected and mock-infected THP-1 and to investigate their small RNA transcriptomes. We found eight miRNAs including miR-US25-1, miR-US25-2-5p and miR-UL112 that were expressed by HCMV during latent infection. The expressions of the host miRNAs were also affected by HCMV latent infection. At least 49 cellular miRNAs were differentially expressed: 39 were up-regulated and 10 were down-regulated upon HCMV latent infection. The expression of the human miRNA hsa-miR-124-3p was significantly up-regulated in the HCMV latent infection library. In addition, we found 14 cellular novel miRNAs in the HCMV-infected and mock-infected THP-1 libraries. Functional annotation of the target genes of the differentially expressed miRNAs suggested that the majority of the genes are involved in melanogenesis, pathways in cancer, endocytosis and wnt signaling pathway.
The small RNA transcriptomes obtained in this study demonstrate the usefulness of the deep-sequencing combined with bioinformatics approach in understanding of the expression and function of host and viral small RNAs in HCMV latent infection. This approach can also be applied to the study of other kinds of viruses.
MicroRNAs（miRNAs）在动植物和病毒的基因表达调控中发挥重要作用。全面描述宿主和病毒 miRNA 将有助于揭示人类巨细胞病毒（HCMV）潜伏感染进展的分子机制。为了研究 HCMV 和潜伏感染宿主细胞中的 miRNA 表达谱，我们对从 HCMV 感染和模拟感染的人单核白血病细胞系 THP-1 中分离的小 RNA 进行了深度测序分析。
我们使用 THP-1 细胞建立了 HCMV 潜伏感染细胞模型。使用高通量测序技术对 HCMV 感染和模拟感染的 THP-1 的小 RNA 文库进行测序，并研究其小 RNA 转录组。我们发现了 8 种 miRNA，包括 miR-US25-1、miR-US25-2-5p 和 miR-UL112，它们在 HCMV 潜伏感染过程中表达。宿主 miRNA 的表达也受到 HCMV 潜伏感染的影响。至少有 49 种细胞 miRNA 在 HCMV 潜伏感染后表达发生变化：39 种上调，10 种下调。HCMV 潜伏感染文库中人类 miRNA hsa-miR-124-3p 的表达显著上调。此外，我们在 HCMV 感染和模拟感染的 THP-1 文库中发现了 14 种细胞新 miRNA。差异表达 miRNA 的靶基因功能注释表明，大多数基因参与黑色素生成、癌症通路、内吞作用和 Wnt 信号通路。
本研究获得的小 RNA 转录组表明，深度测序与生物信息学方法相结合的方法在理解 HCMV 潜伏感染中宿主和病毒小 RNA 的表达和功能方面非常有用。这种方法也可以应用于其他类型病毒的研究。
