在年龄相关性白内障中，miR-211通过靶向沉默信息调节因子2同源物1促进晶状体上皮细胞凋亡。
miR-211 promotes lens epithelial cells apoptosis by targeting silent mating-type information regulation 2 homolog 1 in age-related cataracts.
作者信息
Lu Bo, Christensen Ian T, Ma Li-Wei, Wang Xin-Ling, Jiang Ling-Feng, Wang Chun-Xia, Feng Li, Zhang Jin-Song, Yan Qi-Chang
机构信息
Department of Ophthalmology, the Fourth Affiliated Hospital of China Medical University, Key Laboratory of Lens Research of Liaoning Province, Eye Hospital of China Medical University, Shenyang 110005, Liaoning Province, China.
University of Utah School of Medicine, Salt Lake City, Utah 84132, USA.
出版信息
Int J Ophthalmol. 2018 Feb 18;11(2):201-207. doi: 10.18240/ijo.2018.02.04. eCollection 2018.
AIM
To detect the expression of miR-211 in age-related cataract tissue, explore the effects of miR-211 on lens epithelial cell proliferation and apoptosis, and identify its target gene.
METHODS
This study used real-time quantitative polymerase chain reaction (RT-qPCR) to measure the expression of miR-211 and its predicted target gene [silent mating-type information regulation 2 homolog 1 (SIRT1)] in 46 anterior lens capsules collected from age-related cataract patients. Human lens epithelial cell line (SRA01/04) cells were transfected with either miR-211 mimics, mimic controls, miR-211 inhibitors or inhibitor controls, 72h after transfection, miRNA and protein expression of SIRT1 were measured using RT-qPCR and Western blotting; then cells were exposed to 200 µmol/L HO for 1h, whereupon cell viability was measured by MTS assay, caspase-3 assay was performed. Dual luciferase reporter assay was performed to verify the relationship between miR-211 of SIRT1.
RESULTS
Compared to the control group, expression of miR-211 was significantly increased (<0.001), the miRNA and protein expression of SIRT1 were significantly decreased (<0.001) in the anterior lens capsules of patients with age-related cataracts. Relative to the control group, SIRT1 miRNA and protein levels in the miR-211 mimic group were significantly reduced, cell proliferation activity significantly decreased, and caspase-3 activity was significantly increased (<0.001). In the miR-211 inhibitor group, SIRT1 miRNA and protein expression were significantly increased, cell proliferation activity significantly increased, and caspase-3 activity was significantly decreased (<0.001). A dual luciferase reporter assay confirmed that SIRT1 is a direct target of miR-211.
CONCLUSION
miR-211 is highly expressed in the anterior lens capsules of patients with age-related cataracts. By negatively regulating the expression of SIRT1, miR-211 promotes lens epithelial cell apoptosis and inhibits lens epithelial cell proliferation.
目的
检测miR-211在年龄相关性白内障组织中的表达，探讨miR-211对晶状体上皮细胞增殖和凋亡的影响，并鉴定其靶基因。
方法
本研究采用实时定量聚合酶链反应（RT-qPCR）检测46例年龄相关性白内障患者的前囊膜中miR-211及其预测靶基因[沉默信息调节因子2同源物1（SIRT1）]的表达。将人晶状体上皮细胞系（SRA01/04）细胞分别转染miR-211模拟物、模拟物对照、miR-211抑制剂或抑制剂对照，转染72小时后，用RT-qPCR和蛋白质印迹法检测SIRT1的miRNA和蛋白质表达；然后将细胞暴露于200μmol/L过氧化氢1小时，随后用MTS法检测细胞活力，进行半胱天冬酶-3检测。进行双荧光素酶报告基因检测以验证miR-211与SIRT1之间的关系。
结果
与对照组相比，年龄相关性白内障患者前囊膜中miR-211的表达显著增加（<0.001），SIRT1的miRNA和蛋白质表达显著降低（<0.001）。相对于对照组，miR-211模拟物组中SIRT1的miRNA和蛋白质水平显著降低，细胞增殖活性显著降低，半胱天冬酶-3活性显著增加（<0.001）。在miR-211抑制剂组中，SIRT1的miRNA和蛋白质表达显著增加，细胞增殖活性显著增加，半胱天冬酶-3活性显著降低（<0.001）。双荧光素酶报告基因检测证实SIRT1是miR-211的直接靶标。
结论
miR-211在年龄相关性白内障患者的前囊膜中高表达。通过负调控SIRT1的表达，miR-211促进晶状体上皮细胞凋亡并抑制晶状体上皮细胞增殖。
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