Tang D G, Honn K V
Department of Radiation Oncology, Wayne State University, Detroit, MI 48202, USA.
Adv Exp Med Biol. 1997;400A:349-61. doi: 10.1007/978-1-4615-5325-0_48.
Adherent B16 amelanotic melanoma (B16a) cells exposed to fatty acid 12(S)-HETE, a lipoxygenase metabolite of arachidonic acid, demonstrated a gradual dissolution of stress fibers and bundling-together of vimentin. The 12(S)-HETE effects on tumor cell cytoskeleton appeared 5 min after treatment, became prominent approximately 15 min following stimulation, and generally disappeared by 30 min. Simultaneous treatment of cells with 12(S)-HETE and okadaic acid (OA) prevented disappearance of the 12(S)-HETE effects by 30 min. Quantitative double immunoblotting of actin and vimentin indicated that actin, but not vimentin, underwent a time-related depolymerization. On the other hand, enhanced phosphorylation of vimentin but not of actin was observed after 12(S)-HETE treatment. 12(S)-HETE-enhanced vimentin phosphorylation was abolished by protein kinase C (PKC) inhibitor calphostin C, thus suggesting the involvement of PKC.
暴露于脂肪酸12(S)-HETE（花生四烯酸的脂氧合酶代谢产物）的贴壁B16无黑色素黑色素瘤（B16a）细胞，显示出应激纤维逐渐溶解以及波形蛋白聚集在一起。12(S)-HETE对肿瘤细胞细胞骨架的作用在处理后5分钟出现，刺激后约15分钟变得明显，通常在30分钟时消失。用12(S)-HETE和冈田酸（OA）同时处理细胞可防止12(S)-HETE的作用在30分钟时消失。肌动蛋白和波形蛋白的定量双免疫印迹表明，肌动蛋白而非波形蛋白经历了与时间相关的解聚。另一方面，在12(S)-HETE处理后观察到波形蛋白而非肌动蛋白的磷酸化增强。12(S)-HETE增强的波形蛋白磷酸化被蛋白激酶C（PKC）抑制剂钙磷蛋白C消除，因此提示PKC参与其中。
