The Effects Of ZNF580Overexpression On The Balloon-injury Model Of Rat

Objective：ZNF580(gene ID51157), a novel gene encoding a C2H2(Cys2-His2) zinc-finger transcription factor, may be involved in themaintenance of vascular endothelium homoeostasis. The role of ZNF580insmooth muscle cells is completely unknown. Here, We hypothesized thatoverexpression of ZNF580may inhibit VSMC proliferation,migration andsuppress neointimal hyperplasia in the balloon-injury model of Rat.Methods:1Rat VSMCs were isolated from the aortic media of male Sprague-Dawleyrats. Cells were cultured in Dulbecco Modified Eagle Media (DMEM,Gibco) supplemented with10%fetal bovine serum (FBS),100IU/mLpenicillin, and100IU/mL streptomycin in5%CO2at37℃. For ZNF580overexpression, the recombinant lentivirus vector were prepared byGenePharma.2For overexpression of ZNF580, VSMC were transfected with the lentivirusfor72h.Then, ZNF580expression was monitored by Western blot andquantitative RT-PCR.3Cell proliferation was measured as BrdU incorporation in VSMC grown inpresence of10%FCS for24hours.4Effects of overexpression of ZNF580on VSMCs migration weredetermined by a transwell migration assay5DNA microarray was used to screen the effects of overexpression ofZNF580on the expression profile of VSMC.6Delivered Lentiviral vector expressing ZNF580or a control gene afterballoon injury to investigate the effect of overexpression ZNF580gene onneointima formation. Results:1VSMC can steadily overexpress ZNF580gene after transfected with thelentivirus and evaluated by RT-PCR and Western blot.2Overexpression of ZNF580suppressed the proliferation and migration ofVSMCs.3The results of gene chip inspect shows that161genes significantlymodified by≥2-fold (16up and147down)and718genes weredifferentially expressed by≥1.5-fold (131up and587down)in VSMCthat overexpression of ZNF580gene.The most important altered genesbelonged to functional categories of cell cycle, cell signal transduceractivity were validated by Real Time-PCR.4Overexpression of ZNF580significantly suppressed neointimalhyperplasia at days28after injury in the balloon-injury model of rat.Conclusions:1It was revealed that the overexpression of ZNF580changed theexpression profile of VSMC significantly by means of microarrayanalysis.2The overexpression of ZNF580suppressed the proliferation andmigration of VSMCs in vitro.3The transfer of the ZNF580gene was found to reduce neointimaformation after balloon injury in vivo,These results indicate thatoverexpression of ZNF580may provide a promising strategy for theprevention of restenosis after vascular interventions.