Conditioned media of carcinoma cells cultured in hypoxic microenvironment stimulate angiogenesis in vitro;...

Conditioned media (CM) harvested from human pulmonary squamous cell carcinoma (QG56), pulmonary small cell carcinoma (QG90) and gastric adenocarcinoma (MKN28) cultivated under hypoxic conditions (3% oxygen), enhanced the angiogenic activity in vitro more than those obtained under normoxic cultivation (20% oxygen). The total length of the tube structures formed by bovine capillary endothelial cells (BCEs) in the CM cultured at 3% oxygen was about 1.5 (QG56 and MKN28) or 1.9 (QG90) times longer than that at 20% oxygen. Tube formation was diminished by the preincubation of CM with anti-basic fibroblast growth factor (bFGF) IgG. After performing the fractionations of the CM and the crude extracts of cell lysates cultured using a heparin-Sepharose column, the mitogenic activity in the CM from all cancer cells at 3% oxygen was about twice that of CM at 20% oxygen, while it decreased in the cell lysates at 3% oxygen to about 40% of those at 20% oxygen. This mitogenic activity of BCEs in the CM from all cancer cells was almost totally suppressed by anti-bFGF IgG, but not with anti-vascular endothelial growth factor IgG. Hypoxia is an important factor in tumor angiogenesis by bFGF or bFGF-like molecule(s) derived from tumour cells.