MiRNA-mediated VCP Gene Silence Inhibits The Proliferation And Migration In Human Osteosarcoma Cell Line U2OS In Vitro

Objective：To investigate the effects of VCP silence on the proliferation and migration inhuman osteosarcoma cell line U2OS in vitro.Methods:Three recombinant plasmids (named X135-1-2,X135-2-1,X135-3-1, respectively)targeting VCP gene were constructed, as well as a negative plasmid. All positiveplasmids were identified by bases and then transfected, respectively, to humanosteosarcoma cell line U2OS which expresses VCP at a high level, with negativeplasmid transfecting U2OS cells and normal U2OS cells acting as negative controlgroup and positive control group, respectively. VCP mRNA and protein expressionwere detected by PCR and western blot, respectively; MTT assay was performed toexamine cell proliferation; Transwell Migration Assay was used to evaluate themigration ability of U2OS cells.Result:The levels of VCP mRNA and protein were down-regulated dramatically byrecombinant plasmids targeting VCP, especially the plasmid X135-3-1group,showing a down-regulation of mRNA and protein by (81.4±3.0)%and (83.3±6.0)%,respectively. The activity of cell proliferation in each VCP-knockdown group wasobviously inhibited(P<0.05); In comparison with the control groups, the migrationability of U2OS cells in experimental groups were significantly reduced (P<0.05).The proliferation and migration abilities were reduced some more hardly in plasmidX135-3-1transfecting group compared with another two positive plasmids (X135-1-2, X135-2-1) transfecting groups(P<0.05).Conclusion:VCP gene silence can efficiently inhibit the proliferation and migration of U2OScells in vitro, which has the potential to become a new molecular target forosteosarcoma therapy.