摘要
目的:研究肌动蛋白磷酸化在溶血磷脂酸(LPA)致乳腺癌细胞(MDA-MB-231)肌动蛋白细胞骨架重构及细胞迁移中的作用。方法:10μmol/LLPA孵育MDA-MB-231细胞4h,用Transwell法检测细胞迁移速度;用Western blot及免疫印迹技术检测细胞内骨架组分及胞浆组分中肌动蛋白的分布改变;用二维电泳法分离并比较细胞内骨架组分及胞浆组分中磷酸化和非磷酸化的肌动蛋白含量。结果:10μmol/L浓度的LPA可明显增加MDA-MB-231细胞迁移速度。LPA处理后细胞内F-肌动蛋白含量明显增加,而肌动蛋白总量并未出现明显变化。此外,LPA处理还可明显升高细胞内胞浆组分中磷酸化的肌动蛋白量;在骨架组分中,肌动蛋白仅以磷酸化的形式存在,且含量在LPA刺激前后无明显差异。结论:LPA可促进乳腺癌细胞的迁移能力及细胞骨架发生聚合,其作用可能与其改变细胞胞浆中肌动蛋白的磷酸化水平有关。
Objective：To investigate the effect of actin phosphorylation on the migration ability in human breast （MDA-MB-231） cancer cells and its role in the reorganization of actin cytoskeleton. Methods：The MDA-MB-231 cells were treated with LPA（10 μmol/L） for 4 h. Migration rate was measured by Transwell assay. The contents of G-actin （in cytosolic fraction） and F-actin （in cytoskeletal fraction） in cells were evaluated by Western blotting. Two-dimensional electrophoresis was used to separate phosphorylated actin from unphosphorylated actin in cytosolic and cytoskeletal fraction of cells. Results：After treated with LPA for 4h, the ability of migration in MDA-MB-231 cells was significantly increased compared to the control cells. Consistently,the cellular F-actin was also polymerized after LPA treatment. The proportion of phoshprylated actin after LPA treatment was higher than that of control cells. The further study demonstrated that dephosphorylated actin was only found in cytoplasmic fraction rather than in cytoskletonal fraction. Conclusion： LPA can enhance the ability of migration and polymerization of actin cytoskeleton in MDA-MB-231 cells probably through the phosphorylation of actin.
出处
《南京医科大学学报（自然科学版）》
CAS
CSCD
北大核心
2009年第5期609-612,共4页
Journal of Nanjing Medical University(Natural Sciences)
基金
卫生部基金资助(WKJ2005-2-02)
国家自然科学基金资助(30872926)
