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

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.