E2F圈套寡核苷酸抑制人视网膜色素上皮细胞增生的实验研究  

Inhibitive effect of E2F decoy oligodeoxynucleotide on proliferation of human retinal pigment epithelial cells in vitro

作　　者：李晓青[1] 曾水清[1] 徐莉莉[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》2004年第3期182-185,共4页Chinese Journal of Ocular Fundus Diseases

摘　　要：目的　观察转录因子 E2 F的圈套寡核苷酸 (decoy ODNs)对培养的人视网膜色素上皮(HRPE)细胞增生的影响。　方法　在脂质体 lipofectamine TM2 0 0 0介导下将不同浓度的 E2 F decoy ODNs或错义 decoy ODNs转入培养的 HRPE细胞中 ,甲基四唑盐比色 (MTT)法检测其对 HRPE细胞增生活性的影响 ,凝胶迁移率变动分析 (EMSA)法检测 E2 F decoy ODNs与转录因子 E2 F的竞争结合活性。　结果　 0 .2μmol/L E2 F decoy ODNs显著抑制 HRPE细胞的增生 (P=0 .0 0 2 ) ,并在低浓度时呈剂量依赖关系。错义 decoy则无此作用。EMSA检测结果显示 ,E2 F decoy ODNs拮抗转录因子 E2 F的结合活性。　结论　 E2 F decoy ODNs能序列特异性地抑制转录因子 E2 F的结合活性 ,进而抑制 HRPE细胞的增生活性。Objective To investigate the inhibitive effect of E2F decoy oligodeoxynucleotides (E2F decoy ODNs) on cultured human retinal pigment epithelial (HRPE) cells. Methods E2F decoy ODNs or scramble decoy ODNs at varied concentrations were put into the HRPE cells mediated by lipofectamine TM2000. The proliferative activity of HRPE was detected by methythiazolyl-terazollium assay, and the competitive combinative activity of E2F decoy ODNs and transcription factor E2F was detected by electrophoresis mobility-shift assay. Results The proliferation of HRPE was inhibited markedly by E2F decoy ODNs at the concentration of 0.2 μmol/L (P=0.002) in a dose-dependent manner but not by scrambled decoy. The results of electrophoresis mobility-shift assay showed that the combinative activity of transcription factor E2F was abolished completely by E2F decoy ODNs. Conclusions E2F decoy ODNs may sequence-specifically inhibit the combinative activity of transcription factor E2F,and inhibit the proliferation of HRPE cells.

关 键 词：E2F 人视网膜色素上皮细胞 细胞增生 实验 增生性玻璃体视网膜病变 特异性亲合转录因子 圈套寡核苷酸 

分 类 号：R774.1[医药卫生—眼科]