The response of endothelial cells to TGF beta-1 is dependent upon cell shape, proliferative state and the nature of the substratum.
The response of endothelial cells to TGF beta-1 is dependent upon cell shape, proliferative state and the nature of the substratum.
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发表时间：
1991-08
影响因子：
4
通讯作者：
A. Sutton;A. Canfield;S. Schor;M. E. Grant;A. Schor
中科院分区：
文献类型：
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作者：
A. Sutton;A. Canfield;S. Schor;M. E. Grant;A. Schor
在二维（2-D）基质上培养的内皮细胞增殖，直到它们形成一个紧密贴壁的融合单层静止细胞，显示出典型的“鹅卵石”形态。当加入到增殖培养物中时，TGF β-1（转化生长因子β-1）抑制细胞生长，并引起显著的形态学变化，细胞变大并变得粗糙。这些作用具有剂量依赖性和可逆性。TGF β-1还降低了这些细胞的克隆效率和集落大小，表明TGF β-1对内皮细胞具有细胞毒性和细胞生长抑制作用。相比之下，TGF β-1添加到静止鹅卵石培养物中并不影响细胞形态或细胞数量。在20%血清的存在下，当细胞在2-D基质上培养时，无论细胞是否增殖或鹅卵石静止，TGF β-1以剂量依赖性方式显著增加每个细胞的总蛋白合成水平。纤溶酶原激活物抑制剂1型的水平，特别是在这些文化中增加，所证明的反向纤维蛋白酶谱和免疫沉淀。内皮细胞包埋在3-D胶原蛋白凝胶显示细长的“发芽”形态。这些细胞在凝胶内自我结合形成三维细胞网络，但不增殖。向这些静止的发芽细胞中加入TGF β-1最初诱导了圆整而不改变蛋白质合成，随后发生细胞死亡。TGF β-1对发芽内皮细胞的影响也使用两种培养系统进行了检查，其中鹅卵石和发芽表型都存在。TGF β-1减少了存在的细胞数量和嵌入I型胶原凝胶中的发芽细胞的迁移程度，但对嵌入复杂的内皮产生的细胞外基质中的发芽细胞没有影响。大血管（主动脉）和微血管（视网膜）内皮细胞以类似的方式对TGF β-1作出反应;唯一的区别是在亚融合BREC培养物中未观察到派-1合成增加。我们的研究结果表明，TGF β-1对内皮细胞的影响取决于形状（鹅卵石或发芽），细胞的增殖状态，以及细胞周围基质的性质。这些细胞在体内对TGF β-1的反应可能在血管生成期间通过细胞表型和周围基质组成的变化而类似地调节。
Endothelial cells plated on two-dimensional (2-D) substrata proliferate until they form a tightly apposed confluent monolayer of quiescent cells that display a typical 'cobblestone' morphology. When added to proliferating cultures TGF beta-1 (transforming growth factor beta-1) inhibited cell growth and caused marked morphological changes, with the cells becoming enlarged and ragged. These effects were dose-dependent and reversible. TGF beta-1 also reduced the cloning efficiency and colony size of these cells, indicating that TGF beta-1 is cytotoxic and cytostatic for endothelial cells. By contrast, TGF beta-1 added to quiescent cobblestone cultures did not affect cell morphology or cell numbers. In the presence of 20% serum, the level of total protein synthesis per cell was significantly increased by TGF beta-1 in a dose-dependent manner when the cells were cultured on a 2-D substratum, regardless of whether the cells were proliferating or cobblestone quiescent. The level of plasminogen activator inhibitor type 1 was specifically increased in these cultures, as demonstrated by reverse fibrin zymography and immunoprecipitation. Endothelial cells embedded within a 3-D collagen gel display an elongated 'sprouting' morphology. Such cells self-associate to form three-dimensional cellular networks within the gel, but do not proliferate. The addition of TGF beta-1 to these quiescent sprouting cells initially induced rounding-up without altering protein synthesis, and cell death occurred later. The effects of TGF beta-1 on sprouting endothelial cells were also examined using two culture systems where both the cobblestone and the sprouting phenotypes were present. TGF beta-1 reduced the number of cells present and the extent of migration of sprouting cells embedded within a type I collagen gel, but had no effect upon sprouting cells embedded within a complex endothelial-produced extracellular matrix. Large vessel (aortic) and microvessel (retinal) endothelial cells responded in a similar way to TGF beta-1; the only difference being that an increased synthesis of PAI-1 was not observed with sub-confluent BREC cultures. Our results suggest that the effects of TGF beta-1 upon endothelial cells depend on the shape (cobblestone or sprouting), on the proliferative state of the cells, and on the nature of the matrix surrounding the cells. The response of these cells to TGF beta-1 in vivo may be similarly modulated during angiogenesis by changes in the cell phenotype and the composition of the surrounding matrix.