siMIF抑制人乳腺癌细胞系MCF-7体外迁移和侵袭能力的研究

INHIBITORY EFFECTS OF SIMIF ON THE MIGRATION AND INVASION OF HUMAN BREAST CANCER CELL MCF-7

目的探讨人乳腺癌细胞系MCF-7中巨噬细胞移动抑制因子(macrophage migration inhibitory factor,MIF)的表达受抑制后,对细胞体外迁移和侵袭能力的影响。方法用慢病毒构建可以特异性干涉MIF的小干扰RNA(siMIF)载体,转染人乳腺癌细胞系MCF-7。在倒置荧光显微镜下观察siRNA的转染效率,实时荧光定量PCR检测MIF mRNA表达,Western blot法检测乳腺癌细胞中MIF蛋白和MMP14蛋白表达,细胞迁移实验和Transwell方法检测细胞的体外迁移和侵袭能力。结果慢病毒为载体的siMIF能有效地阻断人乳腺癌细胞系MCF-7的MIF mRNA和蛋白表达,MMP14的表达水平相应降低,且细胞的体外迁移和侵袭能力受到了明显的抑制作用。结论 MCF-7细胞转染特异性siMIF后,其MIF mRNA及蛋白表达被显著抑制,同时细胞的体外迁移和侵袭能力及相关蛋白表达也受到明显抑制。

Objective To investigate the influence of down-regulation of macrophage migration inhibitory factor(MIF)expression on the migration and invasion of MCF-7 cells.Methods Constructed lentivirus-mediated small interfering RNA(siRNA)vectors were transfected into breast cancer cells MCF-7.The efficiency of transfection was observed through an inverted fluorescence microscope.The expression of MIF was measured by the means of real-time fluorescent quantitative polymerase chain reaction;and the expression of MIF and MMP14 protein in breast cancer cells was evaluated by Western blot.The migratory ability of siMIF MCF-7 cells was tested with scar assay and Transwell method.The invasion ability of siMIF MCF-7 cells was tested with Matrigel invasion assay.Results The lentivirus-mediated siMIF blocked the expression of MIF mRNA and protein effectively,as well as MMP14.Besides,the migration and invasion ability of MCF-7 cells was dramatically suppressed by siMIF.Conclusions MIF mRNA and protein are remarkablely restrained after specific siMIF treatment meanwhile siMIF can inhibit the migration and invasion ability of MCF-7 cells.