反义寡核苷酸对视网膜色素上皮细胞中端粒酶逆转录酶表达及细胞增殖的作用  

Effect of antisense oligonucleotides on telomerase reverse transcriptase gene expression and retinal pigment epithe- lium cell proliferation

作　　者：朱宝义[1] 王为农[1] 潘小凤[1] 

机构地区：[1]第四军医大学唐都医院眼科,陕西西安710038

出　　处：《第四军医大学学报》2006年第11期984-986,共3页Journal of the Fourth Military Medical University

摘　　要：目的:研究视网膜色素上皮(RPE)细胞中端粒酶逆转录酶(TERT)的表达及其反义寡核苷酸(ASODN)对其表达和细胞增生的抑制作用,为增生性玻璃体视网膜病变(PVR)探索基因治疗新途径.方法:体外培养兔眼RPE细胞,在不同时间采用链霉亲合素生物素化过氧化物酶复合物(SABC)免疫组织化学法检测TERT的表达;不同浓度的TERTASODN和正义寡核苷酸(SODN)分别作用于体外培养的RPE细胞,采用免疫组织化学方法检测TERT的表达;四唑盐比色法(MTT)检测在不同浓度的TERTASODN和SODN作用下RPE细胞生长活性及其生长抑制率.结果:体外培养兔眼RPE细胞可表达TERT,在5,10μmol/LASODN作用下,TERT的表达明显受抑制;TERTASODN能明显抑制RPE细胞增生活性,并呈剂量依赖性.结论:一定浓度TERTASODN能序列特异性地抑制RPE细胞TERT表达和增生活性,有望进一步用于PVR的治疗实验研究.AIM： To investigate the telomerase reverse transcriptase （TERT） gene expression in retinal pigment epithelium （RPE） cells and the inhibitory effect of antisense oligonucleotides （ASODN） encoding TERT mRNA to gene expression and proliferation of RPE cells, so as to search for a new genetic therapy for proliferative vitreoretinopathy （PVR）. METHODS ： （ 1 ） Rabbit RPE cells cultured in vitro were detected for TERT expression by streptavidin-biotin complex （ SABC ） immunohistochemistry at several time points. （2） The ASODN and sense oligodeoxynucleotides （SODN） encoding TERT were delivered to the RPE cells at different concentrations, and then TERT expression was detected by immunohistochemistry. （3） Exposed to different concentrations of ASODN and SODN, proliferation activity and growthinhibitory rate of RPE cells were measured by MTT methods. RESULTS： TERT expression was markedly suppressed in the RPE cells treated with 5 and 10 μmol/L TERT ASODN. TERT ASODN significantly inhibited proliferative activity of RPE cells in a dose-dependent manner. CONCLUSION： ASODN complementary to TERT mRNA at some concentration can sequencespecifically suppress TERT expression in RPE cells and cellular proliferative activity, and show a potential application in further experimental study for PVR genetic medication.

关 键 词：视网膜色素上皮 寡核苷酸类 反义 端粒酶逆转录酶 

分 类 号：R322.91[医药卫生—人体解剖和组织胚胎学]