THE ESTABLISHMENT OF JAPAN-ORIGINAL GENE THERAPY BY ANTI-TUMOR ANGIOGENESIS USING DECOY GENE.

从人肺鳞状细胞癌（QG56）、肺小细胞癌（QG90）和胃腺癌（MKN28）中获得的条件培养基（CM）在缺氧条件下（3% O2）培养，比在常氧培养（20% O2）下获得的条件培养基（CM）在体外增强血管生成活性。牛毛细血管内皮细胞（BCEs）在3% O2培养的CM中形成的管状结构总长度约为1。5（QG56和MKN28）或1。9（QG90）比20% O2时长。用抗碱性成纤维细胞生长因子（bFGF） IgG预孵育CM可减少管的形成。用肝素- sepharose色谱柱对CM和细胞裂解物粗提物进行分离后，所有癌细胞在3% O2下的CM中有丝分裂活性比在20% O2下的CM高2倍左右，而在3% O2下的细胞裂解物中有丝分裂活性下降到20% O2下的40%左右。抗碱性成纤维细胞生长因子（bFGF） IgG几乎完全抑制所有癌细胞CM中bce的有丝分裂活性，而抗血管内皮生长因子（VEGF） IgG则不具有抑制作用。这些结果提示，缺氧条件是肿瘤细胞衍生的bFGF或bFGF样分子生成肿瘤血管的重要原因。

The 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% O2), enhanced the angiogenic activity in vitro more than those obtained under normoxic cultivation(20% O2). The total length of the tube structures formed by bovine capillary endothelial cells(BCEs) in the CM cultured at 3% O2 was about 1. 5(QG56 and MKN28) or 1. 9(QG90) times longer than that at 20% O2. 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% O2 was about 2 times higher than that in the CM at 20% O2, while it decreased in the cell lysates at 3% O2 to about 40% of those at 20% O2. This mitogenic activity of BCEs in the CM from all cancer cells was almost totally suppressed by anti-basic fibroblast growth factor(bFGF) IgG, but not with anti-vascular endothelial growth factor(VEGF) IgG. These results suggest that the hypoxic condition is an important cause for tumor angiogenesis by bFGF or bFGF-like molecule(s) derived from tumor cells.