Abstract:

Objective     To explore the effect of sense and antisense vascular endothelial growth factor (VEGF) genes on the growth of hypertrophic scar-derived fibroblasts. Methods    Human dermal fibroblasts were isolated from hypertrophic scars by the serum-supplemented media method, and immunocytochemical staining was used to characterize the cell lineage. The genes targeting VEGF including pcDNA3.1 (+)/hVEGF165(the sense group), pcDNA3.1 (-)/hVEGF165(the antisense group) and pcDNA3.1(+)(the empty vector group) were constructed and transfected into human dermal fibroblasts, and a positive clone was selected by G418. The control group was not transfected. Expression of VEGF mRNA was detected by reverse transcription-PCR（RT- PCR）, and expression of the VEGF protein by enzymelinked immunosorbent assay(ELISA)． The growth of dermal fibroblasts was measured by MTT. Results    The eukaryotic expression plasmids of pcDNA3.1(+)/hVEGF165 and pcDNA3.1(-)/hVEGF165 targeting VEGF were successfully constructed. Expressions of VEGF mRNA and protein were obviously increased in the sense group, while they were obviously reduced in the antisense group, compared with those in the empty vector group and control group. The growth lines of human dermal fibroblasts before and after transfection were similar. Conclusion    The sense VEGF gene could promote VEGF expression in human dermal fibroblasts, while the antisense VEGF gene effectively inhibits this expression.

Key words: Hypertrophic scar; Fibroblasts; Vascular endothelial growth factor