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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。
AIM: To investigate the role of actin phosphorylation in actin cytoskeleton remodeling and cell migration in lysophosphatidic acid (LPA) -induced breast cancer cells (MDA-MB-231). Methods: MDA-MB-231 cells were incubated with 10μmol / LLPA for 4h. The migration of cells was detected by Transwell method. The distribution of actin in cytoskeleton and cytoplasm was detected by Western blot and Western blotting. The electrophoretic method was used to separate and compare phosphorylated and non-phosphorylated actin contents in cytoskeletal components and cytoplasmic fractions. Results: LPA at a concentration of 10μmol / L significantly increased the migration rate of MDA-MB-231 cells. LPA treatment significantly increased intracellular F-actin content, while the total amount of actin did not show significant changes. In addition, LPA treatment also significantly increased the amount of phosphorylated actin in the intracellular cytoplasm fraction; actin was only phosphorylated in the cytosolic fraction, and there was no significant difference in the content of LPA before and after stimulation with LPA . CONCLUSION: LPA can promote the migration of breast cancer cells and the cytoskeleton polymerization, which may be related to the changes of actin phosphorylation in cytoplasm.