Isolation and identification of novel Angio genic factors secreted by human ovarian clear cell carcinoma cells and normal bovine pituitary cells

肿瘤组织中存在大量的血管，这些血管为肿瘤组织提供氧气和体液，并使肿瘤组织快速生长，这些血管似乎是由肿瘤细胞通过肿瘤血管生成因子（TAF）诱导形成的。在这项研究中，我们从人卵巢透明细胞癌细胞中分离了新的TAG（非还原条件下MW 78，000，还原条件下MW 82，000）。另一方面，内分泌细胞也可以产生血管生成因子（AGF），因为内分泌腺具有丰富的窦状血管，以传输腺中分泌的激素。因此，我们试图揭示一种新的AGF，这将产生于前垂体腺。结果，我们得到了以下结果：1）牛脑垂体粗提物（Prehormone，Shionogi Pharmaceutical Co.）本实验用免疫组化法检测了层粘连蛋白（laminin，LN）的表达，结果表明，构建血管样小管网络的内皮细胞具有顶侧和底侧极性相反的特点。用牛成纤维细胞培养。构成肾小管的内皮细胞似乎具有正常的顶侧和底侧极性，因为肾小管被层粘连蛋白覆盖。这些结果表明，组成小管的内皮细胞的极性由它们周围的成纤维细胞决定。4）生长激素加速了由垂体粗提物诱导的内皮细胞的小管形成。5）在垂体粗提物的组分中，含有MW 75，000 - 85，000因子的组分诱导了内皮细胞的小管形成。现在，我们正试图揭示这个因子的氨基酸序列。

A cancer tissue has a fund of blood vessels which supply oxygen and mutrition from the host and cause rapid growth of it. These blood vessels are seemed to be induced by cancer cells through tumor angiogenic factors (TAF). In this study, we isolated novel TAG (MW 78,000 non-reduction condition, MW 82,000 under reduction condition) from human ovarian clear cell carcinoma cells. On the other hand, the endocrine cell may also produce an angiogenic factor (AGF) because the endocrine gland has abundant sinusoidal vessels in order to transmit hormones secreted in the gland. Therefore, we have attempted to reveal an novel AGF which would be produced in the anterior pituutary gland. As a result of it, we got the following findings ;1) Crude extracts from the bovine pituitary gland (Prehormone, Shionogi Pharmaceutical Co.) induced monolayr culture cells derived from normal bovine endothelial cells of the pulmonary artery to form networks of blood vessel-like tubules.2) Immunohistochemical stainings of laminin revealed that the endothelial cells constructing the above mentioned networks of tubules have reversed polarity of apical and basal sides.3) The crude pituitary extracts also induced networks of blood vessel-like tubules from bovine endothelial cells co-cultured with bovine fibroblasts. The endothelial cells constructing the tubules seemed to have normal polarity of apical and basal sides because the tubules were covered by laminin. These results suggest that the polarity of the endothelial cells composing the tubules is determined by the fibroblast surrounding them.4) Growth hormone accelerated the tubule-formation of the endothelial cells induced by the crude pituitary extracts.5) Among fractions of the crude pituitary extracts, the tubule formation of the endothelial cells was induced by a fraction containing a factor of MW 75,000-85,000. Now, we are trying to reveal the amino acid sequence of this factor.