Bikfalvi A, Alterio J, Inyang A L, Dupuy E, Laurent M, Hartmann M P, Vigny L, Raulais D, Courtois Y, Tobelem G
Institut des vaisseau et du sang, Hopital Lariboisiere, Paris, France.
J Cell Physiol. 1990 Jul;144(1):151-8. doi: 10.1002/jcp.1041440120.
The human omentum contains a potent, not yet identified angiogenic activity. The omentum is very vascularized. Therefore, we investigated whether human omental microvascular endothelial cells (HOME cells) express the angiogenic peptide basic fibroblast growth factor (bFGF). Cytosol prepared from HOME cells stimulated DNA synthesis in bovine epithelial lens cells (BEL cells). The mitogenic activity could be neutralized by an anti-bFGF antibody. Basic FGF-like material from the HOME cell cytosol was bound onto a heparin-Sepharose column at 0.6 M and was eluted at 3 M NaCl. The 3 M NaCl eluted material reacted with the specific anti-bFGF antibody in an ELISA and stimulated DNA synthesis. It did not react with a specific anti-acidic fibroblast growth factor (aFGF) antibody. Western blotting experiments using the same bFGF antibody showed the presence of a major band of 17 Kd and a doublet of 20-22 Kd. Northern blotting of non-stimulated HOME cells using a specific 1.4 kb bFGF probe showed the presence of 5 molecular species of 6.6, 3.7, 2.2, 2.0, and 1.0 kb. No aFGF mRNA was detected with a specific previously characterized 4.04 kb probe. 12-O-tetradecanoylphorbol 13-acetate (TPA) did not influence significantly the expression of bFGF at the protein and mRNA level in HOME cells. Thus, protein kinase C activation by TPA did not appear to modulate significantly the expression of bFGF for that cell type. Contrastingly, human umbilical vein endothelial cells (HUVE cells), which expressed no bFGF and aFGF mRNA at a basal level, were induced to express bFGF but not aFGF mRNA when stimulated by TPA. These results suggest that the described angiogenic activity could be the bFGF-like mitogen contained in HOME cells and that these cells are different from endothelial cells derived from large vessels (HUVE cells) regarding the expression of bFGF.
人网膜含有一种强大但尚未明确的血管生成活性。网膜血管非常丰富。因此，我们研究了人网膜微血管内皮细胞（HOME细胞）是否表达血管生成肽碱性成纤维细胞生长因子（bFGF）。从HOME细胞制备的胞质溶胶刺激了牛晶状体上皮细胞（BEL细胞）中的DNA合成。这种促有丝分裂活性可被抗bFGF抗体中和。来自HOME细胞胞质溶胶的碱性FGF样物质在0.6M时结合到肝素-琼脂糖柱上，并在3M NaCl时洗脱。3M NaCl洗脱的物质在ELISA中与特异性抗bFGF抗体反应并刺激DNA合成。它不与特异性抗酸性成纤维细胞生长因子（aFGF）抗体反应。使用相同的bFGF抗体进行的蛋白质印迹实验显示存在一条17Kd的主要条带和一条20 - 22Kd的双峰条带。使用特异性1.4kb bFGF探针对未刺激的HOME细胞进行Northern印迹分析显示存在6.6、3.7、2.2、2.0和1.0kb的5种分子种类。用先前鉴定的特异性4.04kb探针未检测到aFGF mRNA。12 - O - 十四烷酰佛波醇13 - 乙酸酯（TPA）对HOME细胞中bFGF在蛋白质和mRNA水平的表达没有显著影响。因此，TPA激活蛋白激酶C似乎并未显著调节该细胞类型中bFGF的表达。相比之下，人脐静脉内皮细胞（HUVE细胞）在基础水平不表达bFGF和aFGF mRNA，在受到TPA刺激时被诱导表达bFGF但不表达aFGF mRNA。这些结果表明，所描述的血管生成活性可能是HOME细胞中所含的bFGF样促有丝分裂原，并且这些细胞在bFGF表达方面与大血管来源的内皮细胞（HUVE细胞）不同。
