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

癌症组织有一个血管基金，这些血管从宿主那里供应氧气和营养物质，并导致宿主的快速生长。这些血管似乎是由癌细胞通过肿瘤血管生成因子(TAF)诱导的。在这项研究中，我们从人卵巢透明细胞癌细胞中分离到了新的TAG(未还原条件下的分子量为78,000，还原条件下的分子量为82,000)。另一方面，内分泌细胞也可能产生血管生成因子(AGF)，因为内分泌腺有丰富的正弦血管，以传输腺体中分泌的激素。因此，我们试图揭示一种新的AGF，它将在脑下垂体前叶产生。结果，我们得到了以下发现：1)牛脑下垂体(前激素，Shionogi Pharmtics Co.)诱导来自正常牛肺内皮细胞的单层培养细胞形成血管样小管网络。2)层粘连蛋白免疫组织化学染色显示，构成上述小管网络的内皮细胞具有顶侧和基底侧的反转。3)垂体粗提物还可诱导与牛成纤维细胞共培养的牛内皮细胞形成血管样小管网络。由于小管被层粘连蛋白覆盖，构成小管的内皮细胞在顶侧和基底侧似乎具有正常的极性。这些结果表明，构成小管的内皮细胞的极性是由它们周围的成纤维细胞决定的。4)生长激素促进了垂体粗提物诱导内皮细胞的小管形成。5)在垂体粗提物的部分中，内皮细胞的小管形成是由一个相对分子质量为75,000-85,000的部分诱导的。现在，我们正试图揭示该因子的氨基酸序列。

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.