Modulation of expression and assembly of vinculin during in vitro fibrillar collagen-induced angiogenesis and its reversal
Modulation of expression and assembly of vinculin during in vitro fibrillar collagen-induced angiogenesis and its reversal
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DOI：
10.1006/excr.1996.0131
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发表时间：
1996-05-01
影响因子：
3.7
通讯作者：
Lapiere, CM
中科院分区：
文献类型：
--
作者：
Deroanne, CF;Colige, AC;Lapiere, CM
胶原诱导的体外血管生成模型被用来研究在分化过程中粘着斑相关蛋白的表达和组装的调节。人脐静脉内皮细胞（HUVEC），首先附着在粘附基质（明胶、纤连蛋白或层粘连蛋白包被的培养皿）或粘附胶原凝胶上，然后被覆盖的胶原凝胶覆盖，在3-4天内组织成管状结构（TLS）。从完全分化的HUVEC中去除覆盖的胶原凝胶诱导了该过程的逆转，HUVEC返回到单层模式。通过Western blot分析研究了HUVEC体外分化过程中发生的粘着斑相关蛋白的调节及其逆转。在TLS中观察到黏着斑蛋白、整合素α 2亚基、talin、α-辅肌动蛋白和肌动蛋白的表达显著降低，而FVIII相关抗原的量与对照单层培养物相比没有变化。在逆转过程中，所有减少的蛋白质显着重新表达。人皮肤成纤维细胞（HSF），提交到相同的实验条件下，没有形成TLS，在HSF中的大多数粘着斑蛋白类似地由覆盖的胶原凝胶调节，除了黏着斑蛋白，它没有被修改。因此，对这种特殊的蛋白质进行了更彻底的研究。在一个未分化的单层HUVEC，一个显着比例的黏着斑被组织成一个洗涤剂耐药的adjacentamembranous结构（局灶性粘附斑块）解体早期TLS形成和重组过程中的逆转。在TLS形成过程中黏着斑蛋白的减少之前，其mRNA的下调，而这种mRNA的上调，在逆转的形态，这些结果表明，细胞骨架和粘着斑蛋白的调制，更具体地说，黏着斑蛋白耦合到其亚细胞再分布是关键的和早期的事件，在体外血管生成过程中的机械化学信号级联由纤维状胶原蛋白诱导。(C)出版社：Academic Press，Inc.
A model of collagen-induced in vitro angiogenesis was used to investigate the modulation of expression and assembly of focal adhesion plaque-associated proteins during the process of differentiation. Human umbilical vein endothelial cells (HUVEC), first attached on an adhesive substratum (gelatin-, fibronectin-, or laminin-coated dish) or adherent collagen gel and then covered by an overlaying collagen gel, organized within 3-4 days in tube-like structures (TLS). Removing the overlaying collagen gel from fully differentiated HUVEC induced a reversion of the process and HUVEC returned to a monolayer pattern. Modulations of focal adhesion-associated proteins occurring in HUVEC during the in vitro differentiation process and its reversal were investigated by Western blot analysis. A significant decrease of expression of vinculin, the integrin alpha(2) subunit, talin, alpha-actinin, and actin was observed in TLS whereas the amount of FVIII-related antigen did not vary as compared to control monolayer cultures. During reversal, all the reduced proteins were markedly reexpressed. Human skin fibroblasts (HSF), submitted to the same experimental conditions, did not form TLS, Most of the focal adhesion proteins in HSF were similarly modulated by an overlaying collagen gel with the exception of vinculin, which was not modified. This particular protein was therefore more thoroughly investigated. In a nondifferentiated monolayer of HUVEC, a significant proportion of vinculin was organized into a detergent-resistant juxtamembranous structure (focal adhesion plaque) which disassembled early in TLS formation and reassembled during the reversal of the process. The reduction of vinculin during TLS formation was preceded by a downregulation of its mRNA while this mRNA was upregulated during reversal of the morphotype, These results suggest that the modulations of the cytoskeletal and focal adhesion proteins and more specifically of vinculin coupled to its subcellular redistribution are critical and early events in the cascade of mechanochemical signaling during in vitro angiogenesis induced by fibrillar collagen. (C) 1996 Academic Press, Inc.