New biological active substances produced by a human malignant tissue

人体恶性组织产生的新生物活性物质

基本信息

批准号：
61480182
负责人：
MATSUZAKI Fukashi
金额：
$ 4.61万
依托单位：
The First Department of Intrnal Medicine, Medical Center. Saitama Medical School
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (B)
财政年份：
1986
资助国家：
日本
起止时间：
1986 至 1987
项目状态：
已结题

项目摘要

We have observed that a human malignant melanoma cell line transplanted to nude mice induced cachexia in the mice. The pathophysiological conditions were very similar to those of cachexia observed in cancer patients. We postulated that the cachexia might be induced by some biological active substances produced and released by the tumor. Since serum glucose levels of the tumor bearing mice decreased and glycogen content of the tumor formed in the nude mice incresed, we tested whether or not the melanoma cells were producing insulin like activity. We found that conditioned media from the cultured melanoma cells contained insulin like activity tested in vitro using rat fat cells. Fractronation by membrane and gel filtration revealed that active substances consisted of at least two components with a molecular weight about 5,000 and 30,000. Hence we could not detect any EGF recepter in the cells, we checked whether the cells were seceting EGF like activity. We could detect high levels of EG … More F like activity in the conditioned media by radiorecepter assay. The physicochemical nature of the EGF like activity was different from those of mouse and human EGF and similar to that of transforming growth factor (TGF)-<alpha>. From various points of view, we concluded that EGF like activity was identical to TGF-<alpha>. We found that the TGF-<alpha> production by the cells was enhanced by retinoic acid. Using collectd dulture media of the cells added tetinoic acid,we succeeded in purification of TGF-<alpha> enough to immunize animals. We could get specific antibodies which did not cross react human and mouse EGF. Using the antibodies, we established a sensitive assay method. By the assay, we could detect TGF-<alpha> in the urine from tumor bearing mice, This suggests that TGF-<alpha> produced by the tumor may be one of causal substances to the cachexia induced by the tumor. We also found that this cell was producing TGF-<bata>. Therefor more than three substances may be related to the induction of cachexia in tumor bearing mice. Less

我们观察到将人类恶性黑色素瘤细胞系移植到裸鼠体内引起恶病质，其病理生理状况与癌症患者中观察到的恶病质非常相似，我们推测恶病质可能是由某些生物活性物质引起的。由于荷瘤小鼠的血清葡萄糖水平下降并且裸鼠中形成的肿瘤的糖原含量增加，我们测试了黑色素瘤细胞是否产生胰岛素样活性。培养的黑色素瘤细胞的条件培养基含有胰岛素样活性，通过膜和凝胶过滤进行体外测试表明活性物质由至少两种分子量约为5,000和30,000的成分组成，因此我们无法检测到任何活性物质。细胞中的 EGF 受体，我们检查了细胞是否分泌 EGF 样活性，我们可以通过放射受体在条件培养基中检测到高水平的 EG F 样活性。 EGF样活性的理化性质不同于小鼠和人EGF，并且与转化生长因子(TGF)-α相似。从不同的角度来看，我们得出结论，EGF样活性与小鼠和人EGF相同。我们发现，使用添加了丁酸的细胞收集的培养基，可以增强细胞产生的TGF-α。 TGF-α足以免疫动物，我们获得了不与人和小鼠EGF发生交叉反应的特异性抗体，通过该测定，我们可以检测到TGF-α。来自荷瘤小鼠的尿液，这表明肿瘤产生的TGF-α可能是肿瘤引起的恶病质的致病物质之一，因此我们还发现该细胞产生超过3种TGF-β。物质可能相关对荷瘤小鼠恶病质的诱导作用较小。