下调CENP-E对乳腺癌MCF-7细胞的影响

Effect of Low Expression of CENP-E on Breast Cancer Cell Line MCF-7

目的:探讨有丝分裂检查点蛋白着丝粒蛋白-E(CENP-E)基因在肿瘤发生发展中的作用。方法:利用shRNA下调CENP-E基因的表达,分别用巢式PCR和Western blot检测CENP-E mRNA和蛋白的表达;MTT检测CENP-E下调后MCF-7细胞的增殖变化;流式细胞术检测CENP-E下调后对MCF-7细胞凋亡的影响;Transwell试验检测MCF-7细胞的迁移和侵袭能力变化;间接免疫荧光检测细胞内CENP-E蛋白和有丝分裂情况。结果:shRNA能有效抑制CENP-E mRNA和蛋白的表达。MTT结果显示CENP-E下调后MCF-7细胞的增殖能力减弱(P<0.05);流式细胞术显示下调CENP-E后能促进MCF-7细胞的凋亡;间接荧光结果显示CENP-E干扰后MCF-7细胞内CENP-E蛋白减少并伴有核分裂异常;Transwell试验显示CENP-E干扰组细胞的迁移和侵袭能力增强(P<0.05)。结论:下调部分CENP-E的表达能抑制MCF-7细胞的增殖,促进MCF-7细胞的凋亡,增强MCF-7细胞的迁移和侵袭能力。

Objective To explore the role of mitotic protein CENP-E in cancer.Methods Plasmid vector containing CENP-E special short hair RNA（shRNA）was transfected into human breast cancer MCF-7 cells.Nest PCR and Western blot was used to detect CENP-E mRNA and protein level;the proliferation of cells with down-regulated CENP-E were analyzed by MTT assay;the apoptosis of cells were detected by flow cytometry（FCM）;the migration and invasion ability of cells were evaluated by transwell assay;indirect immunofluorescence was used to detect CENP-E protein and cell mitosis.Results Compared with the control group,the CENP-E mRNA and protein level in MCF-7 cells transfected shRNA-CENP-E were decreased;the cell growth were inhibited in shRNA-CENP-E group（P0.05）;the results of FCM showed that the cell apoptosis ratio were obviously increased after down-regulated CENP-E;indirect immunofluorescence indicated CENP-E protein was reduced after transfected with shRNA-CENP-E,and the nucleus showed abnormal caryogram;but the cell ability of migration and invasion in shRNA-CENP-E group were increased（P0.05）.Conclusion： Down-regulated part of CENP-E gene expression in breast cancer MCF-7 cells can inhibit cell proliferation,induce cell apoptosis,meanwhile enhance the cell ability of migration and invasion.