YB-1表达对乳腺癌细胞迁移和侵袭能力的影响  

Effects of Y-box binding protein-1(YB-1) on invasiveness and migration capability of breast cancer cells

作　　者：杨靖[1] 许文林[1] 李娟[1] 弓晋灵[1] 吴晓君[1] 陈巧云[1] 王法春[1] 

机构地区：[1]江苏大学附属人民医院中心实验室,江苏镇江212002

出　　处：《江苏大学学报（医学版）》2010年第3期207-211,共5页Journal of Jiangsu University：Medicine Edition

摘　　要：目的:观察Y-盒结合蛋白-1(YB-1)对肿瘤细胞迁移和侵袭能力的影响。方法:通过脂质体介导的方法将YB-1 shRNA重组载体转染入人类乳腺癌MCF-7细胞,经G418筛选及克隆化培养后获得稳定转染株。运用RT-PCR和蛋白质印迹方法检测MCF-7细胞中YB-1 mRNA和YB-1蛋白的表达水平,运用划痕实验和Transwell小室法检测细胞运动、迁移和侵袭能力的变化,同时运用RT-PCR和蛋白质印迹方法检测细胞中MMP-2的表达水平,并用明胶酶谱法检测MMP-2蛋白的活性。结果:稳定转染YB-1 shRNA的细胞中YB-1的表达明显低于转染随机片段的细胞和未转染组细胞,且细胞的运动、迁移和侵袭能力以及MMP-2的表达及活性也较后两组细胞明显下降。转染随机片段的细胞和未转染组细胞间无明显差异。结论:YB-1 shRNA可以沉默乳腺癌MCF-7细胞中YB-1的表达,并降低细胞的迁移和侵袭能力以及MMP-2的表达及活性。Objective： To study the effects of Y-box binding protein-1（YB-1） on the invasiveness and migration capability of breast cancer cells.Methods： The recombinant eukaryotic expression plasmid including YB-1 shRNA was introduced into MCF-7 cells by lipofectamine mediation and the cell line with stable expression of YB-1 was obtained by G418 screening and colony culture.The expression levels of YB-1 mRNA and protein were determined by RT-PCR and Western blotting.The mobility,invasiveness and migration capability of cells were evaluated by using wound healing assay and Transwell chamber model in vitro while the expression levels of MMP-2 mRNA and protein were determined by RT-PCR and Western blotting,and the activity of MMP-2 protein was detected by Gelatin Zymography.Results： The expression of YB-1 in the cells transfected with YB-1 shRNA was much less than the cells transfected with random fragment and the control group.The mobility,invasiveness,migration capability and the expression of MMP-2 as well as the activity of MMP-2 protein of the cells transfected with YB-1 shRNA were much lower than the other two groups while the differences between the other two groups were not significant.Conclusion： The interference of YB-1 with YB-1 shRNA in breast cancer MCF-7 cells can suppress the expression of YB-1 and the invasiveness and migration capability of cells,and decrease the expression of MMP-2 and the activity of MMP-2 protein.

关 键 词：Y-盒结合蛋白-1 MCF-7 SHRNA 迁移和侵袭能力 MMP-2 

分 类 号：R737.9[医药卫生—肿瘤]