Expression of vascular endothelial growth factor in endothelial cells: an in vitro study

Objective To observe the expression and production of vascular endothelial growth factor (VEGF) induced by PMA in a human umbilical vein endothelial derived cell line ECV-304 according to American Tissue and Cell Collection(ATCC) so as to set up a convenient and stable model for the study of endothelial function. Methods RT-PCR was used to detect VEGF mRNA expression, while flow cytometry combined with specific anti-VEGF antibody to measure intracellular VEGF protein production. ELISA was utilized to determine the VEGF concentration secreted by ECV-304. Results It was found that 100 ng/ml PMA, the optimal concentration determined in the present study, increased markedly the VEGF mRNA expression after 2 h incubation. This effect lasted for at least 12 hours and reached the maximum by 12 h Actinomycin D depressed the effect of PMA. The ELISA data demonstrated that the secreted VEGF protein level reached the maximum by 18 h. Conclusions PMA can induce VEGF expression and production in ECV-304 deposeding on dose-and time involving the whole synthetic processing of the VEGF mRNA.