体外转染γ-干扰素基因抑制人眼球筋膜囊成纤维细胞的研究  

Experimental study on the effects of interferon-γ gene on Tenon′s capsule fibroblasts in vitro

作　　者：蓝育青[1] 葛坚[2] 林明楷[2] 郑健樑[2] 魏菁[1] 刘海泉[2] 李艳艳[2] 

机构地区：[1]中山大学附属第二医院眼科,广州510060 [2]中山大学附属第二医院中山眼科中心,广州510060

出　　处：《中华眼科杂志》2005年第6期527-532,共6页Chinese Journal of Ophthalmology

基　　金：国家自然科学基金资助项目(39700153);广东省自然科学基金资助项目(970082);广东省卫生厅基金资助项目(B2001037)

摘　　要：目的探讨阳离子脂质体介导的γ-干扰素(IFN-γ)基因对体外培养的人眼球筋膜囊成纤维细胞增殖的抑制作用。方法采用脂质体介导的方法,以质粒pcDNA3IFNγ基因转染体外培养的人眼球筋膜囊成纤维细胞,同时以空载体重组质粒pcDNA3作为对照,G418筛选抗性克隆并扩增。以RTPCR、免疫组化及流式细胞仪间接免疫荧光法检测IFN-γ基因的表达,应用流式细胞仪进行细胞周期分析,采用四甲基偶氮唑盐法检测IFN-γ基因对成纤维细胞的作用。结果转染pcDNA3IFN-γ基因的成纤维细胞在转录水平和蛋白水平表达IFN-γ基因,转染的IFN-γ基因可明显抑制成纤维细胞的增殖。结论转染的IFN-γ基因对体外培养的人眼球筋膜囊成纤维细胞的增殖具有抑制作用。Objective To investigate the effects of interferon-gamma (IFN-γ) gene on Tenon′s capsule fibroblasts in vitro.Methods Using lipofectAMINE, IFN-γ gene was transferred to human Tenon′s capsule fibroblasts with plasmid pcDNA3IFN-γ in vitro. Cells treated with plasmid pcDNA3 without IFN-γ gene were used as the controls. The transfected fibroblasts were screened by G418. The expression of IFN-γ was determined by RT-PCR, immunohistochemistry and flow cytometry assay. The effects of IFN-γ on the proliferation of Tenon′s capsule fibroblasts were evaluated by flow cytometry and MTT. Results The Tenon′s capsule fibroblasts transferred the IFN-γ gene could express the IFN-γ RNA and protein transiently. The proliferation of the fibroblasts transferred the IFN-γ gene was inhibited.Conclusion The proliferation of the Tenon′s capsule fibroblasts in vitro can be inhibited by transferred IFN-γ gene.

关 键 词：眼球筋膜囊 Γ-干扰素 基因抑制 体外转染 IFN-γ基因 pcDNA3 阳离子脂质体介导 四甲基偶氮唑盐法 成纤维细胞增殖 间接免疫荧光法 流式细胞仪 体外培养 RT-PCR 细胞周期分析 抑制作用 基因转染 重组质粒 抗性克隆 免疫组化 

分 类 号：R346[医药卫生—基础医学]