端粒酶编码区的反义寡核苷酸对人视网膜色素上皮细胞增生的影响

Effect of antisense oligonucleotide of telomerase coding region on proliferation of human cultured retinal pigment epithelial cells

目的探讨不同浓度端粒酶编码区的反义寡核苷酸对人视网膜色素上皮细胞增生活性的影响。方法用不同浓度(0μmol·L-1、5μmol·L-1、10μmol·L-1、15μmol·L-1)端粒酶编码区的反义寡核苷酸处理人视网膜色素上皮细胞24h后,使用MTT法测定视网膜色素上皮细胞增生的抑制率,使用流式细胞仪检测其对视网膜色素上皮细胞周期的影响。结果不同浓度端粒酶编码区反义寡核苷酸处理过的视网膜色素上皮细胞的抑制率随反义寡核苷酸浓度的增高而升高,并将视网膜色素上皮细胞阻滞在G0/G1期。结论端粒酶编码区的反义寡核苷酸对视网膜色素上皮细胞有抑制作用且呈浓度依赖性,这一结论可能成为增生性玻璃体视网膜疾病基因治疗的一个新靶点。

Objective To investigate the inhibitory effect of antisense oligonucleotide of telomerase coding region on the proliferation of human retinal pigment epithelial cells（hRPECs）. Methods The cultured hRPECs were treated by antisense oligonucleotid telomerase at various concentration of 0μmol·L^-1,5μmol·L^- 1, 10μmol·L^-1 and 15μmol·L^-1 for 24 hours.MTT assay was used to detect its inhibition ratio in cultured hRPECs. Flow cytometry was used to analyze cell cycle. Results Inhibiton ratio in cellular growth was positively correlated with the concentration of antisense oligonucleotide of telomerase coding region and the percentage of cells in G0/G1 phase increased from 42.4% to 74.2%, the percentage of cells in G1 and S decreased from 36.4% to 14.9% and from 21.2% to 10.9% respectively（P〈 0.001）.Conclusion Antisense oligonucleotide of telomerase coding region can inhibit the proliferation of hRPECs thus providing a potential, novel gene therapeutic approach to PVR prevention.