MiRNA-494 Regulates Invasion And Migration Of Osteosarcoma Cell By Targeting FGFR2

Objective To investigate the mechanism of small RNA-494 mediated FGFR2 regulation on invasion and migration of osteosarcoma cells.Methods The miR-494 level of osteosarcoma 143B cells was up-regulated or down regulated by transient transfection.MTT assay was used to detect cell proliferation.Transwell was used to detect cell invasiveness.The migration ability of the cells was detected by scratch test,and the expression level of MMP-2 and MMP-9 protein was detected by Western blotting.The target gene of miR-494 was predicted by bioinformatics software Targetscan,and the direct target interaction relationship was verified by double luciferase reporter test.A nude mouse model of osteosarcoma was established in situ,and treated with miR-494 agomir.The effect of up regulation of miR-494 on tumor growth in nude mice was observed.Results The proliferation ability of 143B cell miR-494 was significantly inhibited at all time points,and the cell proliferation ability of osteosarcoma 143B cells was promoted after the down regulation of osteosarcoma cell miR-494.After transient transfection of 24h,the number of cross membrane in the up regulation group of miR-494 was less than that of the control group,but the number of cross membrane in the down regulation group of miR-494 was more than that of the control group.The migration ability of the cells in each group was detected by scratch test.The results showed that the cell migration rate in the up regulation group of miR-494 was less than that of the control group,and the cell migration rate of the down regulation group of miR-494 was more than that of the control group.It is suggested that up regulation of miR-494 can inhibit cell migration,while down-regulation of miR-494 can promote cell migration.The expression level of MMP-2 and MMP-9 protein in miR-494 up-regulated group was lower than that in control group,while the expression level of MMP-2 and MMP-9 protein in miR-494 down regulated group was higher than that in control group.The luciferase activity of FGFR2 wild type and miR-494 mimic co transfection group was significantly lower than that of the control group.Up regulation of miR-494 could inhibit the mRNA level of FGFR2 in osteosarcoma 143B cells,while down regulation of miR-494 could promote mRNA level of 143B cell FGFR2 in osteosarcoma.The tumor volume of miR-494 agomir(494-agomir)drug delivery group in nude mice was smaller than that in control group on the 14,21 and 28 days after administration.The mRNA expression of MMP2,MMP9 and FGFR2 in tumor tissues of 494-agomir group was significantly lower than that of the control group.Conclusion miR-494 can inhibit the proliferation,invasion and migration of osteosarcoma 143B cells,and its mechanism is closely related to the regulation of target gene FGFR2.