Transfection of perlecan by shRNA lentiviral particles on proliferation of fibroblasts in epidural scar

Abstract

Objective To research the proliferation of fibroblasts in epidural scar transfected by perlecan shRNA lentiviral particles.Methods Experimental models were established in 30 Wistar mice which underwent laminectomy.The epidural scar tissue was harvested after culture for 4 days.The fibroblasts in the epidural scar tissue were cultured by tissue explant adherence before they were randomly assigned into no transfection (control group),green fluorescent protein (GFP) lentiviral transfection (GFP group) and shRNA lentiviral transfection (shRNA group).After the fibroblasts were successfully transfected,their expressions of perlecan mRNA and protein and proliferation were detected by Western-blot,RT-PCR and MTT and compared as well among the 3 groups.Results Western-blot and RT-PCR showed the expressions of perlecan mRNA and protein were significantly lower in the shRNA group than in the control and GFP groups (P ＜ 0.05).MTT showed no significant difference among the 3 groups in the optical density at 0 hour (P ＞0.05) but significantly lower optical density in the shRNA group than in the control and GFP groups at 24 to 96 hours (P ＜ 0.05).Conclusion Growth of fibroblasts in epidural scar can be inhibited significantly by transfection with perlecan shRNA lentiviral particles. Key words: RNA; Transfection; Fibroblasts; Cicatrix; Lentivirus