An angiogenic factor from human bladder cancer: the investigation of purification and biological activities.

An angiogenic factor from human transitional cell cancer of bladder has been purified by protein extraction, cation exchange chromatography, gel filtration high-performance liquid chromatography (GE-HPLC) and reversed-phase high-performance liquid chromatography (RP-HPLC). The purified substance was named bladder cancer angiogenic factor (BCAF). Biological activity of the BCAF was assessed by the method of chick embryo chorioallantoic membrane (CAM) assay and 3H-TdR incorporation into DNA in Balb/C 3T3 cells. The BCAF displayed the potent activities of neovascularization in CAM and DNA synthesis in Balb/C 3T3 cells. The ultrastructural features of blood vessels induced by the BCAF were similar to the blood vessels in tumors. The BCAF contained a protein with an approximate molecular weight of 15,000 D which was identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and silver staining. Amino acid compositions of the BCAF were also analysed by acid hydrolysis.