Effects of danazol on endothelial cell function and angiogenesis

Objective To determine the effects of the androgen-like synthetic steroid, danazol, on endothelial cell function and angiogenesis. Design In vitro cell culture models were designed to investigate three critical steps involved in angiogenesis: endothelial cell proliferation, invasion, and tube formation. Setting Research laboratory at a level one trauma center. Patient(s) Commercially available human umbilical vein endothelial cells (HUVEC) were purchased from a pooled patient source. Intervention(s) The HUVEC cells were treated with danazol at concentrations ranging from 1 to 100 μM and evaluated using a series of angiogenesis assays. Main Outcome Measure(s) Absolute cells numbers were quantified using colorimetric methods and fluorescent dyes for cells exposed to danazol in both proliferation assays and invasion chambers. AngiQuant vl.33 software was used to evaluate the formation of capillary-like structures on extracellular matrix gels in the presence of danazol. Result(s) Endothelial cells scrutinized by our in vitro models exhibited decreased proliferation (up to 86%) and tube length (up to 115%) in the presence danazol. Cellular invasion through extracellular matrix, however, was not apparently affected by danazol under the conditions used. Conclusion(s) Danazol interfered with two of the three steps of angiogenesis studied in vitro. This data may help elucidate the mode of action for danazol in vivo.