Tang D G, Diglio C A, Honn K V
Department of Radiation Oncology, Wayne State University, Detroit, MI 48202.
Prostaglandins. 1993 Mar;45(3):249-67. doi: 10.1016/0090-6980(93)90051-8.
12(S)-HETE, a lipoxygenase metabolite of arachidonic acid, has been demonstrated to induce a reversible retraction of vascular endothelial cells (EC). 12(S)-HETE-induced microvascular EC retraction was blocked by a selective protein kinase C inhibitor, calphostin C, but not by the protein kinase A inhibitor, H8. EC exposed to 12(S)-HETE demonstrated a gradual dissolution of actin microfilaments and a decrease of vinculin-containing focal adhesions. The intermediate filaments, vimentin, also underwent extensive reorganization (i.e., filament bundling and enrichment to the cell filapodia) following 12(S)-HETE treatment. In vivo phosphorylation studies revealed that 12(S)-HETE induced a hyperphosphorylation of several major cytoskeletal proteins including myosin light chain, actin, and vimentin. The increased phosphorylation of these cytoskeletal proteins following 12(S)-HETE stimulation was abolished by calphostin C but not by H8. Confluent EC express alpha v beta 3 in focal adhesions at both the cell body and the cell-cell borders. 12(S)-HETE induced a sequential rearrangement of the alpha v beta 3-containing focal adhesions, resulting in a general decrease in alpha v beta 3 integrin receptors, especially in those retracted EC. 12(S)-HETE-induced rearrangement of alpha v beta 3 was inhibited by calphostin C but not by H8. In contrast to alpha v beta 3, confluent EC enrich alpha 5 beta 1 integrin receptors primarily at the cell-cell borders, colocalizing with extracellular fibronectin and cell cortical microfilaments. 12(S)-HETE treatment also disrupted the cell-border distribution pattern of alpha 5 beta 1 as EC retracted, but no distinct alterations (such as time-related redistribution and quantitative differences) in alpha 5 beta 1 were observed.
12(S)-羟二十碳四烯酸（12(S)-HETE）是花生四烯酸的一种脂氧合酶代谢产物，已被证明可诱导血管内皮细胞（EC）发生可逆性回缩。12(S)-HETE诱导的微血管EC回缩被选择性蛋白激酶C抑制剂钙泊三醇C阻断，但未被蛋白激酶A抑制剂H8阻断。暴露于12(S)-HETE的EC显示肌动蛋白微丝逐渐溶解，含纽蛋白的粘着斑减少。中间丝波形蛋白在12(S)-HETE处理后也发生了广泛的重排（即丝束形成并富集到细胞丝状伪足）。体内磷酸化研究表明，12(S)-HETE诱导包括肌球蛋白轻链、肌动蛋白和波形蛋白在内的几种主要细胞骨架蛋白发生过度磷酸化。12(S)-HETE刺激后这些细胞骨架蛋白磷酸化增加被钙泊三醇C消除，但未被H8消除。汇合的EC在细胞体和细胞-细胞边界的粘着斑中表达αvβ3。12(S)-HETE诱导含αvβ3的粘着斑发生顺序重排，导致αvβ3整合素受体总体减少，尤其是在那些回缩的EC中。12(S)-HETE诱导的αvβ3重排被钙泊三醇C抑制，但未被H8抑制。与αvβ3不同，汇合的EC主要在细胞-细胞边界富集α5β1整合素受体，与细胞外纤连蛋白和细胞皮质微丝共定位。12(S)-HETE处理也随着EC回缩破坏了α5β1的细胞边界分布模式，但未观察到α5β1有明显改变（如时间相关的重新分布和定量差异）。
