Westergren-Thorsson G, Persson S, Isaksson A, Onnervik P O, Malmström A, Fransson L A
Department of Medical and Physiological Chemistry, University of Lund, Sweden.
Exp Cell Res. 1993 May;206(1):93-9. doi: 10.1006/excr.1993.1124.
The effects of various glycosaminoglycans (GAGs) on the growth rate of normal fibroblasts and a fibrosarcoma cell line (HT 1080) were examined. Cells were grown in 96-well microplates in the absence or presence of serum mitogens, epidermal (EGF), platelet-derived (PDGF), acidic fibroblast (aFGF), or basic fibroblast growth factor (bFGF). Cell number was measured by using crystal violet to stain cell nuclei (Westergren-Thorsson, G., Onnervik, P.-O., Fransson, L.-A., and Malmström, A. J. Cell. Phys. 147, 523-530, 1991) and also by using a Coulter counter. In the presence of serum mitogens, L-iduronate (IdoA)-rich GAGs, such as dermatan sulfate, heparin, and highly sulfated heparan sulfate, inhibited proliferation of normal cells (25-35%), whereas HT 1080 cells were unaffected or slightly stimulated. Ham's F-12 supplemented with insulin and transferrin but without growth factors was able to support growth of both cell types. Under these conditions, the IdoA-rich GAGs still suppressed growth of normal cells (40-55%), whereas HT 1080 cells again responded poorly. When growth factors were added proliferation of normal fibroblasts was further stimulated, EGF being the most effective. In the presence of either EGF, PDGF, or bFGF, IdoA-rich GAGs had a sustained inhibitory effect on normal fibroblasts (30-50% at concentrations at or above 10 micrograms/ml). However, in the presence of aFGF, both IdoA-rich and IdoA-poor heparan sulfates enhanced growth (nearly twofold after prolonged exposure) suggesting a stabilization of this growth factor. In general, IdoA-rich GAGs appear to inhibit proliferation of normal cells irrespective of the type of growth factor used. Therefore, GAGs are likely to act directly on cell-derived regulatory components, either before or after internalization. As fibrosarcoma cells were much less sensitive to growth inhibition, they may contain altered receptors for GAGs.
研究了各种糖胺聚糖（GAGs）对正常成纤维细胞和一种纤维肉瘤细胞系（HT 1080）生长速率的影响。细胞在96孔微孔板中培养，分别在无血清促有丝分裂原、有表皮生长因子（EGF）、血小板衍生生长因子（PDGF）、酸性成纤维细胞生长因子（aFGF）或碱性成纤维细胞生长因子（bFGF）的条件下培养。通过使用结晶紫对细胞核进行染色（Westergren-Thorsson, G., Onnervik, P.-O., Fransson, L.-A., and Malmström, A. J. Cell. Phys. 147, 523 - 530, 1991）以及使用库尔特计数器来测量细胞数量。在存在血清促有丝分裂原的情况下，富含L-艾杜糖醛酸（IdoA）的GAGs，如硫酸皮肤素、肝素和高度硫酸化的硫酸乙酰肝素，抑制正常细胞的增殖（25 - 35%），而HT 1080细胞不受影响或略有刺激。添加胰岛素和转铁蛋白但无生长因子的哈姆氏F-12培养基能够支持两种细胞类型的生长。在这些条件下，富含IdoA的GAGs仍然抑制正常细胞的生长（40 - 55%），而HT 1080细胞的反应仍然较弱。当添加生长因子时，正常成纤维细胞的增殖进一步受到刺激，其中EGF最为有效。在存在EGF、PDGF或bFGF的情况下，富含IdoA的GAGs对正常成纤维细胞具有持续的抑制作用（浓度在10微克/毫升及以上时为30 - 50%）。然而，在存在aFGF的情况下，富含IdoA和缺乏IdoA的硫酸乙酰肝素均能促进生长（长时间暴露后增长近两倍），表明这种生长因子得到了稳定。总体而言，无论使用何种生长因子，富含IdoA的GAGs似乎都能抑制正常细胞的增殖。因此，GAGs可能在细胞内化之前或之后直接作用于细胞衍生的调节成分。由于纤维肉瘤细胞对生长抑制的敏感性要低得多，它们可能含有改变的GAGs受体。
