Up-regulated expression of miR-181a in HepG2.2.15 cells transfected via full-length hepatitis B virus genome / 解放军医学杂志

ABSTRACT

Objective To identify and analyze the expression of microRNA miR-181a in HepG2.2.15 cell line transfected by full-length hepatitis B virus genome,and explore the potential role of miR-181a engaged in the development of HBV-related liver disease.Methods Based on the previous data from microRNA microarray,miR-181a specific probe was designed and synthesized.The expressive levels of miR-181a in HepG2.2.15 and its parent cell line HepG2 were analyzed using Northern blot analysis.Some putative targets of miR-181a were predicted by computational software RNAhybrid and confirmed by mRNA microarray.One of the targets was selected and flow cytometry analysis was used to further determine the difference of intracellular HLA-A2 level between the two cell lines.Results Northern blot analysis showed that the expressive level of miR-181a in HepG2.2.15 cell was significantly up-regulated compared with that in HepG2 cell.Some putative targets of miR-181a including C8A,IDH1 and HLA-A were predicted and miR-181a might down-regulate the expression of HLA-A gene via partial complement to the 3'-UTR of HLA-A gene.Consistency with the result of mRNA profile microarray,FCM analysis also showed a significantly lower expression of HLA-A2(43.9%)in HepG2.2.15 cell than that in HepG2 cell(96.6%).Conclusion The expressive level of miR-181a in HepG2.2.15 cell is significantly up-regulated and miR-181a might down-regulate its target gene HLA-A,which might be one of the molecular mechanisms that HBV may escape from the immune response and continue replication in hepatocytes.The knowledge is also helpful for understanding the mechanism of HBV-host microRNA interaction.