FIBRONECTIN CONTROLS CAPILLARY ENDOTHELIAL-CELL GROWTH BY MODULATING CELL-SHAPE
FIBRONECTIN CONTROLS CAPILLARY ENDOTHELIAL-CELL GROWTH BY MODULATING CELL-SHAPE
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DOI：
10.1073/pnas.87.9.3579
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发表时间：
1990-05-01
影响因子：
11.1
通讯作者：
INGBER, DE
中科院分区：
文献类型：
--
作者：
INGBER, DE
建立了一个体外系统来研究纤维连接蛋白(FN)在可溶性血管生成有丝分裂原存在的情况下调节毛细血管内皮细胞生长的机制。内皮细胞在含有恒定的、饱和量的碱性成纤维细胞生长因子的化学定义的培养液中培养。然后，通过三种不同的技术以受控的方式改变细胞-FN接触的形成：(I)用增加FN密度的非粘附性细菌培养皿预涂；(Ii)使用可溶性RGD多肽逐渐抑制细胞表面整合素受体与吸附的FN的结合；以及(Iii)FN涂层表面覆盖越来越厚的聚甲基丙烯酸羟乙酯(一种非粘附性聚合物)，以物理上限制细胞访问FN结合部位。随着FN涂层浓度从.apprxeq增加，内皮细胞变得更加伸展和增殖更快。250到.apprxeq。每微米10,000个FN分子。计算机形态计量学分析证实，细胞形状(投射的细胞面积)由FN接触的密度决定，DNA合成水平与细胞扩散的程度密切相关，与用于扰乱细胞黏附的方法无关。相反，当细胞在悬浮液中生长时，FN包被的微球(直径为4.5µm)无论是可溶性FN还是细胞表面结合都不会对生长产生任何影响，细胞扩散被阻止。这些结果表明，FN控制毛细血管内皮细胞的增殖是基于其支持细胞形状依赖于张力的改变的能力--即通过与细胞表面整合素结合和抵抗施加于这些受体的机械负荷。
An in vitro system has been developed to study the mechanism by which fibronectin (FN) regulates capillary endothelial cell growth in the presence of soluble angiogenic mitogens. Endothelial cells were cultured in chemically defined medium containing a constant, saturating amount of basic fibroblast growth factor. Formation of cell-FN contacts was then varied in a controlled fashion by three different techniques: (i) nonadhesive, bacteriological dishes were precoated with increasing densities of FN; (ii) soluble RGD peptides were used to progressively inhibit binding of cell-surface integrin receptors to adsorbed FN; and (iii) FN-coated surfaces were covered with increasingly thick layers of polyhydroxyethylmethacrylate (a nonadhesive polymer) to physically restrict cell access to FN binding sites. Endothelial cells became more extended and proliferated more rapidly as FN coating concentrations were raised from .apprxeq. 250 to .apprxeq. 10,000 FN molecules per .mu.m2. Computerized morphometric analysis confirmed that cell shape (projected cell areas) was determined by the density of FN contacts and that DNA synthetic levels were tightly coupled to the extent of cell spreading, regardless of the method used to perturb cell adhesion. In contrast, neither soluble FN nor cell-surface binding of FN-coated microbeads (diameter, 4.5 .mu.m) hd any effect on growth when cells were grown in suspension and cell spreading was prohibited. These results suggest that FN controls capillary endothelial cell proliferation based on its ability to support tension-dependent alterations of cell shape-i.e., both by binding to cell-surface integrins and by resisting mechanical loads that are applied to these receptors.