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.
出处
《眼科研究》
CSCD
北大核心
2004年第4期347-350,共4页
Chinese Ophthalmic Research
