Inhibition of anchorage-dependent cell spreading triggers apoptosis in cultured human endothelial cells.
Inhibition of anchorage-dependent cell spreading triggers apoptosis in cultured human endothelial cells.
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DOI：
10.1083/jcb.127.2.537
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发表时间：
1994-10
期刊：
影响因子：
--
通讯作者：
Colotta F
中科院分区：
文献类型：
--
作者：
Re F;Zanetti A;Sironi M;Polentarutti N;Lanfrancone L;Dejana E;Colotta F
当在阻止细胞粘附的底物（聚合物聚羟乙基甲基丙烯酸酯、牛血清白蛋白和特氟龙）上培养时，人内皮细胞（EC）迅速失去活力，半衰期约为10小时。死亡的EC表现出凋亡的形态学和生化特征。蛋白质合成抑制剂环己亚胺延缓了悬浮EC细胞的凋亡过程。为了了解悬浮EC的凋亡机制，我们研究了EC与基质蛋白的粘附或整合素的占用是否会影响EC的自杀。EC与低包被浓度的纤维连接蛋白或玻璃体连接蛋白结合，保持圆形，不能组织肌动蛋白微丝，细胞迅速死亡；相比之下，在高底物浓度下，细胞变得扁平，显示肌动蛋白微丝组织，并保持活力。在悬浮的圆形EC中加入饱和量的可溶性玻璃体连接蛋白并没有减少细胞的凋亡过程。最后，当悬浮EC结合Gly-Arg-Gly-Asp- ser包被微珠（约10微珠/细胞），但仍保持圆形时，凋亡过程不受影响。悬液中癌基因转化的EC对细胞死亡和凋亡的敏感性低于正常EC。总之，这些数据表明，细胞附着于基质或整合素结合本身并不足以维持细胞活力，细胞需要经历某种最小程度的形状改变才能存活。因此，调节与细胞外基质的相互作用可以成为控制血管生成的重要目标。
When cultivated on substrates that prevent cell adhesion (the polymer polyhydroxyethylmethacrylate, bovine serum albumin, and Teflon), human endothelial cells (EC) rapidly lost viability with a half-life of approximately 10 h. Dying EC showed the morphological and biochemical characteristics of apoptosis. The apoptotic process of suspended EC was delayed by the protein synthesis inhibitor cycloheximide. To obtain information as to the mechanism involved in the apoptosis of suspended EC, we investigated whether adhesion to matrix proteins or integrin occupancy in EC retaining a round shape may affect EC suicide. EC bound to low coating concentration of either fibronectin or vitronectin, retaining a round shape and failing to organize actin microfilaments, underwent to rapid cell death; by contrast, cells on high substrate concentrations became flattened, showed actin microfilament organization, and retained viability. Addition of saturating amounts of soluble vitronectin to suspended round-shaped EC did not reduce the process of apoptosis. Finally, when suspended EC bound Gly-Arg-Gly-Asp- Ser-coated microbeads (approximately 10 microbeads/cell), yet retaining a round shape, the apoptotic process was not affected. Oncogene- transformed EC in suspension were less susceptible to cell death and apoptosis than normal EC. Overall, these data indicate that cell attachment to matrix or integrin binding per se is not sufficient for maintaining cell viability, and that cells need to undergo some minimal degree of shape change to survive. Modulation of interaction with the extracellular matrix can, therefore, be an important target for the control of angiogenesis.