Selection And Functional Analysis Of HSA-mirnas Involved In A549 Cell Line Infected With A/Yiwu/H1N1 Influenza Virus

Recurrent epidemics caused by Influenza viruses have made and will still continue to cause great damages to public health and economy. Owing to error-prone polymerase and segmented genome of influenza virus, there is common phenomenon of mutations (antigenic drift) and reassortment (antigenic shift) for influenza virus and emergence of new variants or strains. The antigenic shift can lead to a new virus containing a hemagglutinin (HA), one of the major surface glycoproteins of the virus to which humans have no preexisting immunity. And if the virus has the ability to replicate its own genes and transmit among humans, a pandemic happens. An international outbreak of novel swine-origin influenza virus (H1N1) infection during 2009 was such an example. In order to investigate the role of miRNAs in 2009 H1N1 influenza virus infection, a human alveolar epithelial type II cell (HAEC2) line of A549 strain infected with the influenza A H1N1 virus named as A/Zhejiang/DTID-ZJU02/2009(H1N1) was chosen as the investigation subjects. Human microRNA Array v2.0 was used to analyze miRNAs expression in infected A549 cells at different time points postinfection. Bioinformatics were applied to predict targets of miRNA and to analyze singaling pathways of target genes. This investigation can help us understanding the miRNA molecule mechanism of H1N1 influenza A virus infection, and thus establish foundations for diagnosis and treatment. The main contents and results of our research are as follows:1. Degrees of influenza virus A (H1N1) dilution infecting A549 cells were determined. A549 cells were infected with the virus, and growth curve of the virus in A549 cells was drawed out.2. Human MicroRNA Array v2.0 was used to analyze miRNA expression profiles at 0 h, 24 h, 48 h, 72 h postinfection. At 24 h postinfection, 174 changed miRNAs were detected compared with that of 0 h, in which 28 were increased, and 146 were decreased. At 48 h postinfection, 214 changed miRNAs were detected, in which 21 were increased, and 193 were decreased. At 72 h postinfection, 282 changed miRNAs were detected, in which 19 were increased, and 263 were decreased. 3. Targets of the 21 significantly differentially expressed miRNAs were analyzed by bioinformatics, and the targets were enriched by using KEGG and GO. Five key signaling pathways were found out, and they are MAPK, Apoptosis, CREB, JAK_STAT, and TOLL-like receptor signaling pathways. MiRNAs activated APOPTOSIS, CREB and MAPK signaling pathways. A fraction of miRNAs activated JAK_STAT and TOLL_LIKE_RECEPTOR signaling pathways, and another fraction of miRNAs inhibited JAK_STAT and TOLL_LIKE_RECEPTOR signaling pathways,thus suggesting a balance state in host-virus interaction.