端粒酶逆转录酶反义寡核苷酸对视网膜色素上皮细胞增殖的抑制作用  

Antisense oligonucleotides of TERT inhibit proliferation of RPE cells

作　　者：朱宝义[1] 王为农[1] 潘小凤[1] 

机构地区：[1]中国西安第四军医大学唐都医院眼科,710033

出　　处：《国际眼科杂志》2003年第4期29-31,共3页International Eye Science

摘　　要：目的研究视网膜色素上皮(retinalpigmentepithelium,RPE)细胞中端粒酶逆转录酶(telomerasereversetranscriptase,TERT)表达及其反义寡核苷酸(antisenseoligonucleotides,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(proliferativevitreoretinopathy,PVR)治疗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(streptoavidin-biotin-enzymecomplex,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(senseoligodeoxynucleotides,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TERTASODN能序列特异性地抑制RPE细胞TERT表达和增生活性。Aim To investigate telomerase reverse transcriptase (TERT) gene expre ssion in retinal pigment epithelium (RPE) cells and inhibition of antisense olig onucleotides (ASODN) encoding TERT mRNA to gene expression and proliferation of RPE cells, so as to search for new genetic theraputic methods for pro1iferative vitreoretinopathy (PVR). Methods Rabbit RPE cells cultured in vitro were detec ted for TERT expression by streptoavidin-biotin-enzymecomplex (SABC) immunohisto chemistry at different times. The ASODN and sense oligodeoxynucleotides(SODN) en coding TERT were delivered to the RPE cells at different concentrations, then TE RT expression were detected by immunohistochemistry. Exposed to different concen trations of ASODN and SODN, growth activity and suppressive rate of RPE cells we re measured by MTT methods. Results TERT expression were remarkably suppressed in the RPE cells treated with 5 and 10ìmol/L TERT ASODN. TERT ASODN significant ly inhibited proliferative activity of RPE cells in a dose dependent manner. Con clusion ASODN complementary to TERT mRNA can sequence-specifically suppress TER T expression in RPE cells and cellular proliferative activity.

关 键 词：端粒酶逆转录酶 核苷酸 视网膜色素 上皮细胞 RPE 免疫组织化学 手术 

分 类 号：R774.1[医药卫生—眼科]