Effect of Subcellular Localization of Death Domain-associated Protein on Cellular Function of MCF-7 Cells
Objective To explore the expression of death domain-associated protein(DAXX)and its subcellular localization in MCF-7 cells,discussing their effect on cellular function of MCF-7 cells.Methods MCF-7 cells were transfected with LV-DAXX-RNAi-GFP plasmid and LV-DAXX-GFP overexpression lentiviral plasmid,DAXX was knocked down and overexpressed.The expression and subcellular localization of DAXX in MCF-7 cells were detected by nuclear-cytoplasm separation methods and immu-nofluorescence,cell apoptosis and cell cycle were detected by flow cytometry,and cell proliferation was detected by MTT method.Results Both of the nuclear-cytoplasm separation methods and immunofluorescence confirmed that DAXX was mainly located in nucleus of MCF-7 cells.The transfection of LV-DAXX-GFP lentiviral plasmid mainly up-regulated the expression of DAXX in the cytoplasm of MCF-7 cells,while the transfection of LV-DAXX-RNAi-GFP lentiviral plasmid mainly down-regulated the expression of DAXX in the nucleus of MCF-7 cells.Flow cytometry confirmed that down-regulated the expression of DAXX located in nucleus could increase the apoptosis rate of MCF-7 cells,decreasing the cell proliferation,and block the cells in S phase,while up-regulated the expression of DAXX in cytoplasm had no significant effect on cell apoptosis,cell cycle and cell proliferation of MCF-7 cells.Conclusion DAXX is mainly located in the nucleus of MCF-7 cells.The expression of DAXX in the nucleus or cytoplasm may have the opposite effect on the cellular function of MCF-7 cells.
Death domain-associated proteinBreast cancerSubcellular localization
万方数据
维普
