Effects Of MiR-1904 On Biological Characteristics Of Mouse Hepatoma Cells

Pde4d is located on chromosome 13 in mouse,encoding protein PDE4D(phosphodiesterase 4D)which can specificly degrade c AMP,thus plays an important role in cell activities.miR-1904 is one intron-miRNA located on the 8th intron of Pde4 d.Since there has been few study about the target gene and function of miR-1904,a series of experiments were done to analyze the expression in hepatoma cells and to verify the target relationship between miR-1904 and Pde4 d.Subsequently,effects of miR-1904 on biological behavior characteristics of Hepa1-6 cells were studied.Firstly,the target relationship between miR-1904 and Pde4 d was verified.miR-1904 mimics was transfected to overexpress miR-1904.Quantitative PCR and western blot showed that the expression level of Pde4 d both on m RNA and protein decreased after the overexpression of miR-1904 while the expression level of Pde4 d both on m RNA and protein showed significantly increased when miR-1904 was inhibited.Subsequently,the dual-luciferase reporter assay and rescue experiment further confirmed that Pde4 d was the target gene of miR-1904.Then the effects of miR-1904 on biological characteristics of mouse hepatoma cells were studied.By altering the expression of miR-1904 in hepatoma cells,miR-1904 could inhibit the ability of cell proliferation,colony formation,migration and invasion significantly.The result of cell cycle and apoptosis assay showed that miR-1904 affected cell proliferation by inducing cell apoptosis rather than altering cell cycle.In vivo tumorigenesis assay showed the growth rate and the final weight of miR-1904 overexpression tumor decreased significantly,indicating miR-1904 acts as one tumor supressor.In conclusion,the target gene of miR-1904 is Pde4 d.The upregulation of miR-1904 inhibits cell proliferation,migration,invasion,colony formation as well as tumorigenicity of murine hepatoma cells.Besides these,miR-1904 can induce cell apoptosis.These results suggest that miR-1904 may play an important role as a tumor suppressor miRNA in tumor development,which lays a foundation for clarifying the biological function of miR-1904.