血清诱导人视网膜色素上皮细胞中转录因子E2F1的表达及活化  

Expression and activation of transcription factor E2F1 in cultured human retinal pigment epithelial cells induced by serum

作　　者：李晓青[1] 曾水清[1] 徐莉莉[1] 程扬[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院眼科,武汉430022

出　　处：《中华眼底病杂志》2002年第3期224-226,共3页Chinese Journal of Ocular Fundus Diseases

摘　　要：目的　探讨转录因子 E2 F1在人视网膜色素上皮 (retinal pigment epithelium,RPE)细胞中的表达及活化。　方法　培养的人 RPE细胞经同步化后分为两组 :无血清组和含 2 0 %新生牛血清的血清组。采用 Western blot蛋白印迹法检测两组细胞中转录因子 E2 F1蛋白的表达。用凝胶迁移率变动分析法(EMSA)检测其与 DNA的结合活性。　结果　在 RPE细胞核抽提物中检测到相对分子质量为 6 0× 10 3的E2 F1蛋白 ,血清刺激使其表达增加 (P<0 .0 0 1)。EMSA分析示在血清刺激下 ,E2 F1核蛋白与 DNA的结合活性增强。　结论　转录因子 E2 F1存在于人 RPE细胞的细胞核中 ,血清刺激可诱导其表达增加 ,并增强其与 DNA的结合活性 。Objective To explore the expression and activation of transcription factor E2F1 in cultured human retinal pigment epithelium (RPE) cells. Methods Cultured human RPE cells were divided into two groups after synchronization: one was cultured in Dulbecco′s modified Eagle′s medium (DMEM) without serum; the other was cultured in DMEM supplemented with 20% serum of newborn calf. The expressions of E2F1 protein in two groups were detected by Western blot analysis. The E2F1 DNA binding activities were measured by gel mobility shift assay(EMSA). Results E2F1 protein of 60 000 molecular weight was detected in the nuclear extract of human RPE cells, and serum stimulation could increase its expression( P <0.001). EMSA exhibited the increased binding activity of E2F1 in the serum stimulated RPE cells with DNA. Conclusions E2F1 is expressed in the nuclei of human RPE cells. Serum stimulation can increase its protein expression as well as binding activity ,so as to play a regulation role of gene transcription.

关 键 词：血清 人视网膜色素上皮细胞 转录因子 E2F1 病理 增生性玻璃体视网膜病变 

分 类 号：R774.1[医药卫生—眼科]