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

从人肺鳞癌（QG 56）、肺小细胞癌（QG 90）和胃腺癌（MKN 28）中收获的条件培养基（CM）在低氧条件（3%O2）下培养，其体外血管生成活性比在常氧条件（20%O2）下获得的条件培养基更强。牛毛细血管内皮细胞（BCEs）在3%O2培养条件下形成的管状结构的总长度约为1。5（QG 56和MKN 28）或1。9（QG 90）倍于20%O2时。管形成减少CM与抗碱性成纤维细胞生长因子（bFGF）IgG的预孵育。在对CM和使用肝素-琼脂糖柱培养的细胞裂解物的粗提取物进行分级分离后，在3%O2下来自所有癌细胞的CM中的促有丝分裂活性比在20%O2下的CM中的促有丝分裂活性高约2倍，而在3%O2下的细胞裂解物中的促有丝分裂活性降低至在20%O2下的促有丝分裂活性的约40%。抗碱性成纤维细胞生长因子（bFGF）IgG几乎完全抑制了所有癌细胞CM中BCE的促有丝分裂活性，但抗血管内皮生长因子（VEGF）IgG未抑制BCE促有丝分裂活性。这些结果表明，缺氧条件是肿瘤细胞来源的bFGF或bFGF样分子引起肿瘤血管生成的重要原因。

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.