Objective To investigate the expression of vascular endothelial growth factor-A (VEGF-A) and the phenotypic transition after the activation of fibroblasts by the supernatant of cultured tumor cells.Methods The growth tendency of fibroblasts was tested by the MTT assay.The expressions of alpha-smooth muscle actin (α-SMA) and VEGF-A mRNA were tested by RT-PCR.The expressions of α-SMA and VEGF-A protein were tested by immunohistochemistry and Western blot.Results The MTT assay indicated that the conditional medium which contained tumor cells supernatant could obviously promote the growth of the fibroblasts. RT-PCR and Western blot manifested that α-SMA expressed by the fibroblasts which cultured by normal medium reached its peak on day 5,then decreased to a low level on day 7.When the medium contained 2 ng/ml transforming growth factor-β1 (TGF-β1),the fibroblasts could steadily express more α-SMA.But the above two mediums could not make the fibroblasts express the VEGF-A. When using the conditional medium,the α-SMA peak advanced on the third day and maintained at a high level,so as the expression of the VEGF-A.Conclusions The results suggested that fibroblasts can be activated to be myofibroblasts when using the conditional medium.The best activation time of the fibroblasts is consistent with the time of the VEGF-A expression at the highest level by the activated fibroblasts.The fibroblasts which activated at the best time are expected to become a kind of cells which can be used for promoting revascularization.