Objective:To investigate the inhibitory effects of adenovirus-mediated NDRG2 on the proliferation of human renal cell carcinoma cell line OS-RC-2 in vitro.Methods:NDRG2 was harvested by RT-PCR,confirmed by DNA sequenc...Objective:To investigate the inhibitory effects of adenovirus-mediated NDRG2 on the proliferation of human renal cell carcinoma cell line OS-RC-2 in vitro.Methods:NDRG2 was harvested by RT-PCR,confirmed by DNA sequencing,and then cloned into the eukaryotic expression vector pIRES2-EGFP,which encodes green fluorescent protein(GFP),to construct p1RES2-EGFP-NDRG2 plasmid.OS-RC-2 cells with NDRG2 negative expression were transfected with p1RES2-EGFP-NDRG2 plasmid.The growth of transfected OS-RC-2 cells was observed under light and fluorescence microscopes.After colony-forming cell assays,cell proliferation detection and MTT assays,the growth curves of cells in each group were plotted to investigate the inhibitory effects of adenovirus-mediated NDRG2 on the proliferation of OS-RC-2 cells.Cell cycle was determined by flow cytometry.Confocal laser scanning microscopy showed that NDRG2 protein was specifically located on subcellular organelle.Results:A eukaryotic expression vector p1RES2-EGFP-NDRG2 was successfully constructed.After NDRG2 transfection,the growth of OS-RC-2 cells was inhibited.Flow cytometry showed that cells were arrested in S phase but the peak of cell apoptosis was not present,and confocal laser scanning microscopy showed that NDRG2 protein was located in mitochondrion.Conclusions:NDRG2 can significantly inhibit the proliferation of OS-RC-2 cells in vitro and its protein is specifically expressed in the mitochondrion.展开更多
基金supported by Jiangsu Province Natural Science Foundation of China(js2817298)
文摘Objective:To investigate the inhibitory effects of adenovirus-mediated NDRG2 on the proliferation of human renal cell carcinoma cell line OS-RC-2 in vitro.Methods:NDRG2 was harvested by RT-PCR,confirmed by DNA sequencing,and then cloned into the eukaryotic expression vector pIRES2-EGFP,which encodes green fluorescent protein(GFP),to construct p1RES2-EGFP-NDRG2 plasmid.OS-RC-2 cells with NDRG2 negative expression were transfected with p1RES2-EGFP-NDRG2 plasmid.The growth of transfected OS-RC-2 cells was observed under light and fluorescence microscopes.After colony-forming cell assays,cell proliferation detection and MTT assays,the growth curves of cells in each group were plotted to investigate the inhibitory effects of adenovirus-mediated NDRG2 on the proliferation of OS-RC-2 cells.Cell cycle was determined by flow cytometry.Confocal laser scanning microscopy showed that NDRG2 protein was specifically located on subcellular organelle.Results:A eukaryotic expression vector p1RES2-EGFP-NDRG2 was successfully constructed.After NDRG2 transfection,the growth of OS-RC-2 cells was inhibited.Flow cytometry showed that cells were arrested in S phase but the peak of cell apoptosis was not present,and confocal laser scanning microscopy showed that NDRG2 protein was located in mitochondrion.Conclusions:NDRG2 can significantly inhibit the proliferation of OS-RC-2 cells in vitro and its protein is specifically expressed in the mitochondrion.
