miR-210靶向沉默Bcl-2诱导人晶状体上皮细胞凋亡

MiR-210 induced human lens epithelial cell apoptosis by target silencing Bcl-2

目的观察miR-210在年龄相关性白内障晶状体组织中的表达,并探讨其对人晶状体上皮细胞凋亡的影响与调控机制。方法收集年龄相关性白内障患者与新鲜人眼透明晶状体前囊膜(正常对照组)标本;培养人晶状体上皮细胞SRA01/04,予或不予(正常对照组)紫外线照射,利用实时荧光定量PCR检测上述组织或细胞中miR-210表达。此外,将遭受紫外线照射的SRA01/04细胞分为空白对照组、阴性对照组、miR-210模拟物组和miR-210抑制物组,分别予培养液及miR-210阴性对照物、模拟物、抑制物处理48 h,行实时荧光定量PCR检测miR-210表达,以验证转染效率;Western blot检测miR-210的靶基因Bcl-2表达,流式细胞术分析细胞凋亡率。结果与正常对照组相比,年龄相关性白内障患者晶状体前囊膜组织和紫外线诱导的SRA01/04细胞凋亡模型中miR-210表达均显著上调(均为P<0.01)。相对于空白对照组,miR-210模拟物组miR-210水平和细胞凋亡率均增加(均为P<0.01),Bcl-2蛋白表达水平降低(P<0.01);而miR-210抑制物组miR-210水平和细胞凋亡率均减少(均为P<0.05),Bcl-2蛋白表达水平升高(P<0.01);阴性对照组上述指标与空白对照组比较,差异无统计学意义(P>0.05)。结论 miR-210在年龄相关性白内障患者晶状体组织中呈高表达,miR-210可能通过靶向沉默Bcl-2促进人晶状体上皮细胞凋亡。

Objective To observe the expression of miR-210 in lens tissues of age-related cataract,and explore its effects on human lens epithelial cell apoptosis and regulating mechanisms. Methods The anterior lens capsules of age-related cataract patients and fresh human eye transparent lens（ normal control group） were collected. After culturing,human lens epithelial cell line SR A01 /04 was treated with or without（ normal control group） ultraviolet irradiation. Fluorescence real time quantitative PC R was used to detect miR-210 expression in the tissues and cells mentioned above. In addition,SR A01 /04 cells exposed to ultraviolet were randomly divided into blank control group,negative control group,miR-210 mimic group and miR-210 inhibitor group. These cells were then treated with culture medium,control miR-210,miR-210 mimic and miR-210 inhibitor for 48 hours,respectively. Expression of miR-210 was then measured by real time quantitative PC R to validate transfection efficiency. W estern blot was used to examine the expression of Bcl-2,a target gene of miR-210. Apoptotic rate of SR A01 /04 cells was also analyzed using flow cytometry. Results C ompared with normal control group,miR-210 expression was significantly increased in the anterior lens capsules of age-related cataract patients and ultraviolet-induced apoptosis model of SR A01 /04 cells（ all P 〈 0. 01）. C ompared with blank control group,miR-210 levels and apoptotic rate were significantly increased（ all P〈 0. 01）,but Bcl-2protein expression levels were significantly decreased in miR-210 mimic group（ P〈 0. 01）,however,a marked reduction of miR-210 levels and apoptotic rate as well as a marked enhancement of Bcl-2 protein expression levels were seen in miR-210 inhibitor group（ all P〈 0. 01）. There was no significant difference in above indicators betw een negative control group and blank control group（ P〉 0. 05）. Conclusion MiR-210 is highly expressed in lens tissues of age-related cataract patients,and miR-210 may directly target Bcl-2 and then promote apoptosis of human lens epithelial cells.