EphrinA2基因促进MDA-MB-231乳腺癌细胞迁移的初步研究

目的 在MDA-MB-231人乳腺癌细胞系中过表达EphrinA2基因,研究其对肿瘤细胞迁移能力的影响,并初步探讨其可能的作用机制。 方法 用脂质体法将EphrinA2真核表达质粒转染至MDA-MB-231细胞中,用Western blot法检测转染前后细胞中EphrinA2、E-Cadherin及p27/Kip1蛋白的表达;并用细胞疏散、细胞划痕及Transwell小室试验检测转染前后细胞迁移能力的变化;用Rac1抑制剂NSC23766抑制Rac1活性后用细胞划痕试验检测细胞迁移能力变化。 结果 转染EphrinA2后的细胞迁移能力更强,并且呈现出E-Cadherin及p27/Kip1蛋白表达下降;用NSC23766抑制Rac1活性能抑制EphrinA2介导的细胞迁移。 结论 EphrinA2能促进MDA-MB-231乳腺癌细胞的迁移,其机制可能与E-Cadherin和p27/Kip1蛋白下调及Rac1活性抑制有关。

A Primary Study on EphrinA2 Promoted Migration of MDA-MB-231 Breast Cancer Cells

Objective To investigate the effect of EphrinA2 on cancer cell migration by transfecting recombinant eukaryotic expressing vector containing EphrinA2 cDNA into MDA-MB-231 human breast cancer cells and to explore its potential mechanism. Methods We transfected recombinant eukaryotic expressing vector containing EphrinA2 cDNA into MDA-MB-231 cells by lipofectamine 2000. Expression levels of EphrinA2, E-Cadherin and p27/Kip1 protein were measured by Western blotting. Cell migration ability was evaluated by cell scattering, wound healing assay and Transwell migration assay. We used NSC23766 to inhibit Rac1 activity and the cell migration ability after Rac1 inhibition was tested by wound healing assay. Results Cells transfected with EphrinA2 showed enhanced migration ability accompanied with decreased E-Cadherin and p27/Kip1 protein expression. Inhibition of Rac1 by NSC23766 could inhibit EphrinA2-mediated cell migration. Conclusion These results collectively suggest that EphrinA2 could promote MDA-MB-231 breast cancer cell migration and this function possibly involves E-Cadherin and p27/Kip1 protein down-regulation and Rac1 activity inhibition.