Purification and characterization of a novel vascular endothelial cell growth inhibitor secreted by macrophage-like U-937 cells.
Purification and characterization of a novel vascular endothelial cell growth inhibitor secreted by macrophage-like U-937 cells.
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巨噬细胞样 U-937 细胞分泌的新型血管内皮细胞生长抑制剂的纯化和表征。
DOI：
10.1093/oxfordjournals.jbchem.a022296
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发表时间：
1999
期刊：
影响因子：
--
通讯作者：
N. Taniguchi
中科院分区：
文献类型：
--
作者：
N. Nakano;S. Higashiyama;S. Takashima;N. Tsuruoka;M. Klagsbrun;N. Taniguchi
人组织细胞性淋巴瘤细胞系U-937细胞分泌几种血管内皮细胞生长抑制剂，包括白血病抑制因子、制瘤素M、肿瘤坏死因子-α和转化生长因子-β 1。从佛波酯处理的U-937细胞的条件培养基的部分纯化的馏分的表征表明未知的内皮生长抑制剂的存在。使用铜亲和力、肝素亲和力、阳离子交换和反相液相色谱的组合，从条件培养基中将内皮细胞生长抑制剂纯化至均一。在还原和非还原条件下，纯化的生长抑制剂在SDS-聚丙烯酰胺凝胶电泳上迁移为65 kDa条带。生长抑制剂的胰蛋白酶片段的微测序分析和BLAST搜索分析揭示了与已知蛋白质没有同源性的独特序列。纯化的蛋白质以剂量依赖性方式抑制内皮细胞生长，但对平滑肌细胞生长没有影响。该因子还阻断了由成纤维细胞生长因子-2和血管内皮生长因子诱导的内皮细胞生长，并且在通过U-937细胞衍生的内皮细胞生长抑制剂抑制内皮细胞生长方面具有相加效应。因此，该因子似乎是一种新的血管内皮细胞特异性抑制剂，在本研究中暂称为内皮细胞抑制剂（ECI）。
A human histiocytic lymphoma cell line, U-937 cells, secretes several vascular endothelial cell growth inhibitors including leukemia inhibitory factor, oncostatin M, tumor necrosis factor-alpha, and transforming growth factor-beta1. Characterization of partially purified fractions from the conditioned media of phorbol ester-treated U-937 cells suggested the existence of unknown endothelial growth inhibitors. Using a combination of copper affinity, heparin affinity, cation exchange, and reversed phase liquid chromatographies, a growth inhibitor for endothelial cells was purified to homogeneity from conditioned media. The purified growth inhibitor migrated as a 65 kDa band on SDS-polyacrylamide gel electrophoresis under both reducing and nonreducing conditions. Microsequencing analyses of the tryptic fragments of the growth inhibitor and a BLAST search analysis revealed a unique sequence showing no homology to known proteins. The purified protein inhibited endothelial cell growth in a dose-dependent manner, but had no effect on smooth muscle cell growth. The factor also blocked endothelial cell growth induced by both fibroblast growth factor-2 and vascular endothelial growth factor, and was additively effective in inhibiting the growth of endothelial cells by U-937 cell-derived endothelial cell growth inhibitors. Thus, this factor appears to be a novel inhibitor with specificity for vascular endothelial cells, and is tentatively called endothelial cell inhibitor (ECI) in this study.