Bcl-2反义寡核苷酸抑制视网膜色素上皮细胞增生的实验研究  

Inhibition of hRPE cells proliferation by bcl-2 AS-ODN

作　　者：华振宇[1] 赵宏[1] 朱冬梅[1] 高延庆[1] 张蕾[1] 李铮[1] 

机构地区：[1]郑州市第五人民医院卓美眼科中心,450003

出　　处：《眼科研究》2004年第4期347-350,共4页Chinese Ophthalmic Research

摘　　要：目的　探讨不同浓度的Bcl 2反义寡核苷酸 (bcl 2AS ODN)对人视网膜色素上皮 (RPE)细胞增生活性的影响。方法　使用不同浓度 ( 0、15、3 0、45、60 μmol/L)的bcl 2AS ODN和bcl 2S ODN处理人RPE细胞 2 4h后 ,使用MTT、氚 -标记脱氧胸苷掺入试验 ( 3 H TdR)测定RPE细胞增生的抑制率及DNA合成抑制率 ;使用流式细胞仪检测其对RPE细胞周期的影响 ;使用免疫组织化学染色法检测不同浓度bcl 2AS ODN作用下的RPE细胞bcl 2的表达。结果　不同浓度bcl 2AS ODN处理过的RPE细胞的抑制率和DNA合成抑制率随bcl 2AS ODN浓度的增高而升高 ,bcl 2AS ODN将RPE细胞阻滞在G0 /G1期 ,免疫组化结果显示 :bcl 2AS ODN可抑制RPE细胞bcl 2的表达。结论　bcl 2AS ODN对RPE细胞的增生有抑制作用且呈浓度依赖性 ,这一结果有可能成为增生性玻璃体视网膜病变 (PVR)基因治疗的一个新靶点。ObjectiveTo investigate the inhibition of bcl-2 AS-ODN on the proliferation of retinal pigment epithelium (RPE) cells.MethodsThe cultured human RPE cells were treated with bcl-2 AS-ODN at various concentration of 0 μmol/L,15 μmol/L,30 μmol/L, 45 μmol/L and 60 μmol/L for 24 hours,and then quantity of OD and CMP reflecting live cell number and cellular DNA synthesis were respectively measured by methyl thiazolyl tetrazolium (MTT) and tritium-labelled thymidine dexoxyribose( 3H-TdR)incorporation.Flow cytometry was used to analyze cell cycle,and SABC staining was used to detect the expression of bcl-2 in hRPE cells treated with bcl-2 AS-ODN.ResultsInhibiting rate of cellular growth and DNA synthesis were positively correlated with the concentration of bcl-2 AS-ODN,and the percentage of cells in G 0/G 1phase increased from 43.3% to 68.3%.SABC staining demonstrated that expression of bcl-2 showed gradual decrease with an increase in bcl-2 AS-ODN dosage.ConclusionBcl-2 AS-ODN can inhibit the proliferation of hRPE cells thus providing a potential, novel gene therapeutic approach to PVR prevention.

关 键 词：BCL-2反义寡核苷酸 视网膜色素上皮细胞 MTT 氚-标记脱氧胸苷掺人试验 流式细胞技术 免疫组 织化学染色方法 

分 类 号：R774[医药卫生—眼科]