The Inhibitive Effect of Transfection of Eukaryotic Bicistronic Expression Vector pGFAP-IRES2-EGFP-p27 on Astrocytic Proliferation in vitro
ZHU Zhou▲,PAN Deng-ji,XU Yi,YU Zhi-yuan,XU Sha-bei,WANG Wei.▲Department of Neurology,Tongji Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430030,China
Objective: The effect of transfected liposome-mediated eukaryotic bicistronic expression vector pGFAP-IRES2-EGFP-p27 on astrocytic proliferation was explored in vitro.Methods: The plasmid pGFAP-IRES2-EGFP-p27 was transfected to cultured astrocytes with liposome,then the expressions of EGFP,p27 and Ki67 in astrocytes were observed by immunocytochemistry. Results: Fluorescent microscopy revealed that the expression of EGFP could be detectable at 24 h after the plasmid was transfected to the astrocytes,and the expression of EGFP peaked at 48-72 h.Meanwhile,expression of p27 increased significantly in EGFP-positive cells,while the rate of Ki67-positive cells was much lower in EGFP-positive cells than that in EGFP-negative cells(P0.01). Conclusions: The vectors containing the GFAP promoter resulted in EGFP and p27 expression in astrocytes.EGFP can be a reporter gene to indicate the expression of p27.Astrocytic proliferation can be inhibited significantly by transduction of exogenous gene p27.