Preparation of aniti-tumor tissue endothelium antibodies and its application of cancer-missle therapy

抗肿瘤组织内皮抗体的制备及其在肿瘤导弹治疗中的应用

基本信息

批准号：
07457615
负责人：
MAYUMI Tadanori
金额：
$ 1.28万
依托单位：
Osaka University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1995
资助国家：
日本
起止时间：
1995 至 1996
项目状态：
已结题

项目摘要

The permeation of macromolecular FITC-labeled dextran (Mw=70,000) through bovine aortic endothelial cells (BAEC) monolayr, which were cultured for 5 days with conditioned medium prepared from mouse melanoma B16, was increased. However, when BAEC,which were cultured with normal medium until confluent, were treated with B16 conditioned medium (B16-CM) for 30 min, the permeability did not increase. The B16-CM also increased the permeability of the endothelial monolayrs of bovine veins and the human umbilical vein, but did not increase that of the epithelial monolayr. The B16-CM did not alter the distribution or content of F-actin on the BAEC.BAEC cultured in the presence of B16-CM for 5 days were detached from the dish, and then seeded into a chamber at one-fifth of confluent cell density. After 5 days of culture in nomal medium, the BAEC were grown to confluence and their permeability was increased. These findings suggest that B16-CM increased the endothelial permeability irreversibly wi … More thout the decrease of F-actin, and that soluble factor (s) which were secreted from the tumor cells participate in the construction of the hyperpermeable structure of tumor vessels in vivo.Rat KMT-17 fibrosarcoma-derived endothelial cells were isolated by Percoll gradient centrifugation with an attaching-speed separation technique, and their properties in culture were examined. The primary cultured tumor-derived endothelial cells (TEC) showed angiotensin-converting enzyme activity, positivity for Factor VIII-related antigen staining, and typical capillary-like formation on Matrigel. The primary cultured TEC monolayr showed greater permeability than normal tissue-derived endothelial cell (aorta, vena cava and epididymal fat capillary) monolayrs on FITC-dextran diffusion (molecular weight 70,000). Leukocyte adhesion to TEC was reduced compared to that to fat-derived capillary endothelial cells. These characteristics resembled those of tumor vascular endothelium, and were observed both in the primary and first-passage cell cultures, but not in the fourth-passage cell cultures. Our findings indicate that primary or subcultured TEC are applicable for studies of the physiological characteristics of tumor endothelial cells. Less

与小鼠黑色素瘤B16制备的条件培养基一起培养5天的牛主动脉内皮细胞(BAEC)单层中，大分子FITC标记的葡聚糖(Mw=70,000)的渗透性增加，但是，当BAEC与用小鼠黑色素瘤B16制备的条件培养基一起培养时，其渗透性增加。正常培养基至汇合后，用B16条件培养基（B16-CM）处理30分钟，通透性没有变化。 B16-CM还增加了牛静脉和人脐静脉的内皮单层的通透性，但没有增加上皮单层的通透性。B16-CM没有改变F-肌动蛋白在上皮单层上的分布或含量。将在 B16-CM 存在下培养 5 天的 BAEC.BAEC 从培养皿中分离出来，然后以汇合细胞密度的五分之一接种到室中。在正常培养基中培养 5 天后，BAEC 生长至汇合，并且其通透性增加。这些发现表明，B16-CM 不可逆地增加了内皮通透性，但 F-肌动蛋白和可溶性因子 (s) 减少。）由肿瘤细胞分泌，参与体内肿瘤血管高通透结构的构建。通过分离方法分离大鼠KMT-17纤维肉瘤来源的内皮细胞采用附着速度分离技术进行 Percoll 梯度离心，并检查了原代培养的肿瘤源性内皮细胞 (TEC) 的特性，显示出血管紧张素转换酶活性、因子 VIII 相关抗原染色呈阳性以及典型的毛细血管。原代培养的 TEC 单层显示出比正常组织来源的内皮细胞（主动脉、腔静脉和附睾脂肪）更大的渗透性。与脂肪来源的毛细血管内皮细胞相比，FITC-葡聚糖扩散的白细胞（分子量为 70,000）的单层细胞对 TEC 的粘附减少。我们的研究结果表明，原代或传代培养的 TEC 适用于肿瘤生理特征的研究。内皮细胞较少。