中文摘要：

目的：探讨脂质体介导真核表达载体pGFAP-IRES2-EGFP-p27转染体外培养星形胶质细胞后对其增殖的影响。方法：利用脂质体将质粒pGFAP-IRES2-EGFP-p27转染入纯化培养的星形胶质细胞，通过免疫荧光化学标记观察转染质粒pGFAP-IRES2-EGFP-p27后星形胶质细胞内外源基因EGFP、p27的表达情况，并观察其对星形胶质细胞Ki67表达的影响。结果：脂质体介导质粒pGFAP-IRES2-EGFP-p27转染后24h即开始有EGFP的表达，48-72h EGFP表达达高峰；通过免疫荧光标记发现，表达EGFP的细胞同时有p27表达水平明显增高；和EGFP阴性细胞相比，EGFP阳性细胞中Ki67的阳性率明显下调（P〈0．01）。结论：GFAP启动子能启动目的基因p27和EGFP在星形胶质细胞的表达；连于p27的下游EGFP可作为报告基因，通过观察EGFP的表达可了解外源性p27的表达情况；导入外源性p27能有效抑制星形胶质细胞的增殖。

英文摘要：

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 （P〈0.01）. 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.