转染Zn^(2+)诱导表达反义bcl-2基因对人视网膜色素上皮细胞的影响  

Effect of transfection with Zn^(2+) induced expression of anti-sense bcl-2 gene on human retinal pigment epithelial cells

作　　者：刘洪雷[1] 王雨生[1] 惠延年[1] 

机构地区：[1]中国西安第四军医大学西京医院眼科,710032

出　　处：《国际眼科杂志》2004年第3期404-408,共5页International Eye Science

基　　金：中国国家自然科学基金资助项目(No.39700154) ~~

摘　　要：目的:观察转染Zn2+诱导表达的反义bcl-2 基因对人视网膜色素上皮(RPE)细胞的影响。方法:培养的正常RPE细胞与转染反义bcl-2 RPE细胞采用免疫组化,克隆形成试验、MTT,流式细胞仪,TUNEL分别予以观察。结果:转染后RPE细胞细胞骨架角蛋白表达阳性;硫酸锌诱导表达的安全剂量为160μmol/L;诱导表达反义bcl-2 RPE大部分细胞bcl-2 表达为阴性,部分弱阳性;在第10d细胞克隆形成率分别为(4.65±0.043)%与(4.85±0.085)%(P >0.05),诱导表达反义bcl-2 的RPE生长曲线在5d后表现为低增生率(P <0.01);柔红霉素作用后G2期细胞由9.9%增加至30.4%;180μg/L柔红霉素作用反义bcl-2转染组TUNEL阳性细胞可见典型凋亡小体。结论:转染可诱导表达的bcl-2 基因的RPE细胞可以进行细胞凋亡的后续研究。AIM: To evaluate the effect of transfection with Zn2+ induced expression of anti-sense bcl-2 gene on human retinal pigment epithelial cells.· METHODS: The cultured human retinal pigment cells (RPE) and anti-sense bcl-2 transfected RPE were examined by immunohistochemical assay, clone formation test, tetrazolium (MTT) colorimetric assay, flow cytometry and TUNEL.· RESULTS: The framework of normal RPE cells and transfected RPE cells were stained with keratine antibody; safe induciable dosage of ZnSO4 was 160μmol/L bcl-2 expression level of RPE cells were negative in general and partially trace positive by immunohistochemical assay; cloning efficiency of normal and transfected RPE cells were (4.65±0.043) % and (4.85±0.085)% at 10d respectively (P >0.05); cells growth curve showed lower proliferation ratio of induced anti-sense bcl-2 RPE cells after 5d (P <0.01); the percentage of cells in G2 increased from 9.9% to 30.4%; typical apoptosis body in situ were observed in TUNEL staining positive cells treated with daunomycin wtih the dosage of 180μg/L.· CONCLUSION: RPE cells transfected with Zn2+ induciable anti-sense bcl-2 gene are available for the preceeding research of apoptosis.

关 键 词：反义bcl-2基因 转染 人视网膜色素上皮细胞 免疫组化 柔红霉素 细胞凋亡 

分 类 号：R774.1[医药卫生—眼科]