The Effect Of PcDNA3.1/EGR-1 On Proliferation And Apoptosis Of Ovarian Cancer Cell SKOV3

PART IObjective: To construct the plasmid pc DNA3.1/EGR-1 and import it into human ovarian cancer SKOV3 cells.Methods: Construct the plasmid pc DNA3.1/EGR-1 then transient transfection the recombinant plasmid pc DNA3.1/EGR-1 in human ovarian cancer cells by cationic liposome method.Results: Enzyme the plasmid pc DNA3.1/EGR-1 identify its successfully constructed.The transfection rate of human ovarian cancer cell by cationic liposome method under the fluorescence microscope showed 30-40%.Conclusion: By cationic liposome transfection method, recombinant plasmid pc DNA3.1 / EGR-1 can be more efficient transfection.PART IIObjective:To observe the proliferation and apoptosis of human ovarian cancer cells by the effects of pc DNA3.1/EGR-1.Methods: The effects of EGR-1 expression on cell proliferation, cell apoptosis were analyzed by MTT, Annexin V- FITC/ PI flow cytometry.Results: Compared with the control cells,the proliferation of human ovarian cancer cells transfected with EGR-1 was significantly inhibited( P<0.05),the cell apoptotic rate was increased.Conclusion: EGR-1 inhibits the proliferation of human ovarian cancer cells, increases the apoptotic rate.PART IIIObjective: To detect the expression of EGR-1 gene in human ovarian cancer cells.Methods: Transient transfection the recombinant plasmid pc DNA3.1/EGR-1 in human ovarian cancer cells by cationic liposome method. The expression level of EGR-1 m RNA was detected by real-time quantitative PCR. The expression level of EGR-1 protein was detected by Western blotting.Results: Compared with the control cells, the high expression of EGR-1 was detected by real-time quantitative PCR and Western blotting in the transfected human ovarian cancer cells.Conclusion: the expression of EGR-1 gene is higher in the transfection group than the control group.