权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Purification of cartilage-derived anti-tumor factor (CATF) and establishment of cell lines which produce CATF

软骨源性抗肿瘤因子（CATF）的纯化和产生CATF的细胞系的建立

基本信息

批准号：
61480387
负责人：
TAKIGAWA Masaharu
金额：
$ 4.16万
依托单位：
Osaka University
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

1. Purification of cartilage-derived anti-tumor factor (CATF)CATF was extracted from fetal bovine cartilage with 1 M guanidine hydrochloride and parrially purified by acetone fractionation (45 - 65%), ultrafiltration (Molecular weight; 100-300k) and DEAE-Sepharose CL-6B column chromatography (eluted with 0.3 to 0.35 M NaCl at pH 8.0). The partially purified CATF strongly inhibited the growth of solid sarcoma 180 and B16 melanoma in vivo, the proliferation and DNA synthesis of bovine pulmonary artery endothelial (CPAE) cells in culture and B16 melanoma-induced angiogenesis in chick embryo chorioallantoic membranes (CAM). The specific activity of this partially purified CATF was about 70 times that of crude CATE when determined using its inhibitory action on DNA synthesis in CPAE cells in culture.2. Production of CATF by rabbit costal chondrocytes in primary culture.Serum-free medium conditioned by exposure to rabbit costal chondrocytes in primary culture specifically inhibited the proliferation and DBA synthesis of CPAE cells in Culture. The factor in conditioned medium (CM) also inhibited angiohenesis in CAM induced by B16 melanoma and growth of the tumor transplanted onto the CAM. These findings strongly suggest that rabbit costal chondrocytes produce CATF.3. Establishment of cell lines producing CATF.A cell line showing some phenotypes of growth cartilage, such as responsiveness to parathyroid hormone and high alkaline phosphatase activity was established from secondary cultures of mouse costal growth cartilage. Moreover, a cell line showing typical cartilage phenotypes was established from human chondrosarcoma. CM obtained from these cells inhibited the proliferation and DNA synthesis of CPAE cells in culture. The inhibition was specific for CPAE cells. Therefore, these cell lines are thought to produce CATF.

1.软骨源性抗肿瘤因子（CATF）的纯化用1M盐酸胍从胎牛软骨中提取CATF，并通过丙酮分级（45-65％）、超滤（分子量；100-300k）和DEAE-Sepharose CL-6B柱层析（用0.3至0.35M NaCl洗脱）进行部分纯化 pH 值 8.0）。部分纯化的 CATF 强烈抑制体内实体肉瘤 180 和 B16 黑色素瘤的生长、培养物中牛肺动脉内皮 (CPAE) 细胞的增殖和 DNA 合成以及 B16 黑色素瘤诱导的鸡胚绒毛尿囊膜 (CAM) 中的血管生成。利用其对培养的CPAE细胞中DNA合成的抑制作用测定，该部分纯化的CATF的比活性约为粗CATE的70倍。2．原代培养中兔肋软骨细胞产生CATF。原代培养中暴露于兔肋软骨细胞的无血清培养基特异性抑制培养物中CPAE细胞的增殖和DBA合成。条件培养基（CM）中的因子还抑制B16黑色素瘤诱导的CAM中的血管生成和移植到CAM上的肿瘤的生长。这些发现强烈表明兔肋软骨细胞产生CATF.3。产生CATF的细胞系的建立。从小鼠肋生长软骨的继代培养物中建立了显示生长软骨的一些表型的细胞系，例如对甲状旁腺激素的反应性和高碱性磷酸酶活性。此外，从人软骨肉瘤中建立了显示典型软骨表型的细胞系。从这些细胞中获得的 CM 抑制培养物中 CPAE 细胞的增殖和 DNA 合成。该抑制作用对 CPAE 细胞具有特异性。因此，这些细胞系被认为产生 CATF。