Objective To investigate the effect of over-expression of E2 protein GFP-E2,GFP-TAD(N-extremity domain of HPV18 E2)and GFP-DBD(C-extremity domain of HPV18 E2)fusion proteins on the apoptosis of macrophages in uterine ...Objective To investigate the effect of over-expression of E2 protein GFP-E2,GFP-TAD(N-extremity domain of HPV18 E2)and GFP-DBD(C-extremity domain of HPV18 E2)fusion proteins on the apoptosis of macrophages in uterine cervix cancer was studied.Methods TAD or DBD gene was amplified by PCR from pEGFP-C1/HPV18 E2,and cloned into pEGFP-C1 vector.Then the subclone of pEGFP-C1/TAD or pEGFP-C1/DBD was screened respectively.Forty-eight hours after transfection of pEGFP-C1/HPV18 E2,pEGFP-C1/TAD,pEGFP-C1/DBD or pEGFP-C1 into macrophages,the localization or expression of them was analyzed by fluorescent microscope or western blot,respectively.The apoptotic rate of macrophages was detected by flow cytometry.Results The plasmid of pEGFP-C1/TAD or pEGFP-C1/ DBD was constructed respectively.Macrophages transfected with different plasmid,over-expression of GFP-E2 or GFP-TAD fusion protein up-regulated apoptotic rate of macrophages.Furthermore,the effect of GFP -TAD was more powerful than GFP-E2.But the GFP-DBD over-expressed had not the same effects.Conclusion Over-expression of GFP-E2 or GFP-TAD fusion protein could induce apoptosis of macrophages.展开更多
基金supported by the Edueation Committee of Hunan province(No.02C391)
文摘Objective To investigate the effect of over-expression of E2 protein GFP-E2,GFP-TAD(N-extremity domain of HPV18 E2)and GFP-DBD(C-extremity domain of HPV18 E2)fusion proteins on the apoptosis of macrophages in uterine cervix cancer was studied.Methods TAD or DBD gene was amplified by PCR from pEGFP-C1/HPV18 E2,and cloned into pEGFP-C1 vector.Then the subclone of pEGFP-C1/TAD or pEGFP-C1/DBD was screened respectively.Forty-eight hours after transfection of pEGFP-C1/HPV18 E2,pEGFP-C1/TAD,pEGFP-C1/DBD or pEGFP-C1 into macrophages,the localization or expression of them was analyzed by fluorescent microscope or western blot,respectively.The apoptotic rate of macrophages was detected by flow cytometry.Results The plasmid of pEGFP-C1/TAD or pEGFP-C1/ DBD was constructed respectively.Macrophages transfected with different plasmid,over-expression of GFP-E2 or GFP-TAD fusion protein up-regulated apoptotic rate of macrophages.Furthermore,the effect of GFP -TAD was more powerful than GFP-E2.But the GFP-DBD over-expressed had not the same effects.Conclusion Over-expression of GFP-E2 or GFP-TAD fusion protein could induce apoptosis of macrophages.
