Effect Of Low Expression Of CENP-E On Breast Cancer Cell Line MCF-7

Objective: To explore the role of mitotic protein CENP-E in cancer.Methods: Plasmid vector containing CENP-EWT mRNA special shorthair RNA（shRNA）was transfected into human breast cancer MCF-7cells.Nest PCR was used to detect CENP-EWT mRNA level; and theproliferation of cells with down-regulated CENP-E were analyzed by MTTassay. The apoptosis of cells were detected by flow cytometry（FCM）; themigration and invasion ability of cells were evaluated by transwell assay.Antisense oligonucleotide special for CENP-EI was designed tosilence CENP-EI, and transfected into MCF-7cells. Nest PCR was used todetect CENP-EWT and CENP-EI mRNA level. Real Time PCR wasspecially used to measure CENP-EI mRNA; the proliferation of cells withsilenced CENP-EI were analyzed by MTT assay. Cells colony-formingassay and transwell assay were carried out to detect the changes of invasionand migration in the migration and invasion ability of the cells.Results Compared with the control group, the CENP-E mRNA levelin MCF-7cells transfected shRNA-CENP-EWT were decreased; the cellgrowth were inhibited in shRNA-CENP-EWT group（P<0.05）.The results of FCM showed that the cell apoptosis ratio were obviously increased afterdown-regulated CENP-EWT; but the cell ability of migration and invasionin shRNA-CENP-EWT group were increased(P<0.05).Antisense oligonucleotide special for CENP-EI can effectively inhibitthe expression of CENP-EI mRNA, compared with sense group and blankgroup, meanwhile the CENP-EWT mRNA was not effected. The cellgrowth were enhanced in antisense oligonucleotide group(P<0.05). Thecell colony formation rate of experimental group was （21.67±1.2）%，increased versus the control group（9.6±1.45）%(P<0.05).and the cell abilityof migration and invasion in Antisense oligonucleotide group wereincreased(P<0.05).Conclusion: Down-regulated part of CENP-EWT gene expression inbreast cancer MCF-7cells can inhibit cell proliferation, induce cellapoptosis, meanwhile enhance the cell ability of migration and invasion.While silencing CENP-EI mRNA can enhance the cells growth, increasethe cell colony formation rate and the ability of migration and invasion.