Src激酶抑制剂PP2对乳腺癌MDA-MB-231细胞黏附、迁移能力的影响

Effects of Src inhibitor PP2 on the adhesion and migration of human breast cancer cells(MDA-MB-231)

目的:研究Src激酶抑制剂PP2对人乳腺癌细胞(MDA-MB-231)肌动蛋白细胞骨架及黏附、迁移能力的影响。方法:不同浓度Src激酶抑制剂PP2(0.2、1.0、5.0和25.0!mol/L)孵育MDA-MB-231细胞24h。用FITC标记的鬼笔环肽标记F-肌动蛋白,用荧光显微镜分析肌动蛋白细胞骨架的重组情况;用免疫印迹技术检测细胞内骨架组分(Triton不溶组分)及胞浆组分(Triton可溶组分)中肌动蛋白的分布;划痕损伤实验检测细胞迁移速度;四甲基偶氮唑盐比色法(MTT)检测细胞对人工重组基底膜(Matrigel)的黏附能力。结果:PP2处理可导致细胞内肌动蛋白细胞骨架发生解聚,F-肌动蛋白排列和分布及细胞形态发生明显改变;此外,PP2可明显降低MDA-MB-231细胞迁移速度及其对人工重组基底膜的黏附能力,其效应呈剂量依赖性。结论:Src激酶抑制剂PP2可影响乳腺癌细胞的黏附及迁移能力,其作用可能与其对肌动蛋白细胞骨架的重组有关。

Objective：To investigate the effects of Src signal pathway on the adhesion and migration in human breast cancer cells （MDA-MB-231 ） and its role in the reorganization of actin cytoskeleton. Methods ：The MDA-MB-231 cells were treated with Src inhibitor PP2 in various concentrations（0.2, 1.0,5.0 and 25 μmol/L） for 24 h. MTI： assay was used to detect the adhesive ability of MDA-MB- 231 cells to artificial basement membrane. Migration rate was measured by wound healing assay. To detect the reorganization of actin cytoskeleton,the contents of G-actin （in cytosolic fraction） and F-actin （in cytoskeletal fraction） in cells were evaluated by Western blot, and the distribution of F-actin stained by FITC-phalloidin in cells was also examined by fluorescence microscopy. Results： After treated with PP2 for 24 h,the ability of adhesion and migration in MDA-MB-231 cells were significantly reduced compared to the control cells in a dose-dependent manner. Consistently,the celluar F-actin was also depolymerized after PP2 treatment. Conclusion： Src inhibitor PP2 can inhibit the ability of adhesion and migration in MDA-MB-231 cells, which probably be through the reorganization of actin cytoskeleton.