Purification and characterization of angiogenic factor from human bladder cancer

An angiogenic factor from human transitional cell cancer of bladder was 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 as bladder cancer angiogenic factor (BCAF). Biological activity of the BCAF was assessed by using 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.