Weich H A, Iberg N, Klagsbrun M, Folkman J
Department of Surgery, Children's Hospital, Boston, Massachusetts 02115.
Growth Factors. 1990;2(4):313-20. doi: 10.3109/08977199009167026.
The expression and synthesis of acidic and basic fibroblast growth factors (aFGF and bFGF) in cultures of bovine and human vascular smooth muscle cells (BSMC and HSMC) was studied. BSMC express and synthesize only bFGF, whereas HSMC express and synthesize both bFGF and aFGF. The presence of bFGF in BSMC is shown by the following criteria: (1) the growth factor activity in BSMC lysates binds to a heparin-affinity column and elutes as a single peak at 1.5-1.7 M NaCl, characteristic for bFGF; (2) this extract is mitogenic for smooth muscle cells; (3) Northern blot analysis demonstrates three distinct bFGF mRNAs of 7.0, 4.0, and 1.9 kb; no aFGF mRNA species were detected. Analysis of human umbilical vein endothelial cells yielded similar results: Heparin-affinity chromatography and Northern blot analysis failed to demonstrate the presence of aFGF despite the detection of bFGF by these techniques. In contrast, HSMC synthesize two growth factor activities: First, they bind to an immobilized heparin column and elute as two separate peaks at 1.2 and 1.5-1.7 M NaCl, characteristic for aFGF and bFGF; and second. Northern blot analysis demonstrates the expression of aFGF mRNA of 4.6 kb and bFGF mRNAs of 7.0, 4.0 and 1.9 kb. Furthermore, it is shown that aFGF and bFGF are potent mitogens for smooth muscle cells in vitro.
研究了牛和人血管平滑肌细胞（BSMC和HSMC）培养物中酸性和碱性成纤维细胞生长因子（aFGF和bFGF）的表达与合成。BSMC仅表达和合成bFGF，而HSMC则表达和合成bFGF与aFGF。通过以下标准证明BSMC中存在bFGF：（1）BSMC裂解物中的生长因子活性与肝素亲和柱结合，并在1.5 - 1.7M NaCl处作为单峰洗脱，这是bFGF的特征；（2）该提取物对平滑肌细胞具有促有丝分裂作用；（3）Northern印迹分析显示有7.0、4.0和1.9kb三种不同的bFGF mRNA；未检测到aFGF mRNA种类。对人脐静脉内皮细胞的分析得出类似结果：尽管通过这些技术检测到了bFGF，但肝素亲和色谱法和Northern印迹分析未能证明aFGF的存在。相比之下，HSMC合成两种生长因子活性：第一，它们与固定化的肝素柱结合，并在1.2和1.5 - 1.7M NaCl处作为两个单独的峰洗脱，这分别是aFGF和bFGF的特征；第二，Northern印迹分析显示4.6kb的aFGF mRNA以及7.0、4.0和1.9kb的bFGF mRNA的表达。此外，研究表明aFGF和bFGF在体外是平滑肌细胞的有效促有丝分裂原。
