Regulation mechanism of miR-204 targeted SIRT1 in human lens epithelial cell apoptosis

中国实用眼科杂志 Pub Date : 2017-05-10 DOI:10.3760/CMA.J.ISSN.1006-4443.2017.05.022

Xiao-Liang Xie, Jiangyue Zhao, Fangkun Zhao, Yu Qin

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Abstract

Objective To investigate the effect of miR-204 expression on apoptosis of human lens epithelial cell line (LECs) SRA01/04 in vitro and the mechanism of targeting SIRT1. Methods Hydrogen peroxide (H2O2) was used to construct the cell apoptosis model of human lens epithelial cell line SRA01/04 in vitro. MiR-204 mimic and inhibitor were transiently transfected into SRA01/04 cells using Lipofectamine 2000 to regulate the expression level of miR-204. Flow cytometry was used to detect the cell apoptosis rate in each group SRA01/04 cells. By microRNA databases and recent study researches, we predicted that SIRT1 maybe a potential target gene of miR-204. Quantitative real-time PCR (qRT-PCR) were used to detect the expression level of miR-204 and SIRT1 gene. Results Quantitative real-time PCR results showed that the expression level of miR-204 in cell apoptosis model of SRA01/04 was significantly higher than the normal SRA01/04 cells. Overexpression of miR-204 in SRA01/04 promoted H2O2-induced cell apoptosis, and down-regulated the expression level of SIRT1 gene. While downexpression of miR-204 in SRA01/04 reduced H2O2-induced cell apoptosis, and up-regulated the expression level of SIRT1 gene. Each result was statistically significant (P <0.01). Conclusions The expression level of miR-204 in cell apoptosis model of SRA01/04 is significantly higher than the normal SRA01/04 cells. MiR-204 can regulate the apoptosis of the human lens epithelial cells by targeting SIRT1 gene. Thus, miR-204 may play an important role in the pathogenesis of caracts with lens epithelial cell apoptosis. Key words: miR-204; SIRT1; Cell apoptosis; Cataract

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miR-204靶向SIRT1调控人晶状体上皮细胞凋亡的机制

目的探讨miR-204在体外表达对人晶状体上皮细胞系SRA01/04细胞凋亡的影响及其靶向SIRT1的机制。方法采用过氧化氢(H2O2)体外构建人晶状体上皮细胞株SRA01/04细胞凋亡模型。用Lipofectamine 2000将MiR-204 mimic和inhibitor瞬时转染到SRA01/04细胞中，调节MiR-204的表达水平。流式细胞术检测各组SRA01/04细胞的细胞凋亡率。通过microRNA数据库和近期的研究，我们预测SIRT1可能是miR-204的潜在靶基因。采用实时荧光定量PCR (qRT-PCR)检测miR-204和SIRT1基因的表达水平。结果实时荧光定量PCR结果显示，miR-204在SRA01/04细胞凋亡模型中的表达水平明显高于正常SRA01/04细胞。在SRA01/04中过表达miR-204促进h2o2诱导的细胞凋亡，下调SIRT1基因的表达水平。而miR-204在SRA01/04中下调表达，可减少h2o2诱导的细胞凋亡，上调SIRT1基因的表达水平。各结果均有统计学意义(P <0.01)。结论miR-204在SRA01/04细胞凋亡模型中的表达水平明显高于正常SRA01/04细胞。MiR-204可通过靶向SIRT1基因调控人晶状体上皮细胞的凋亡。因此，miR-204可能在晶状体上皮细胞凋亡的白内障发病机制中发挥重要作用。关键词:miR-204;SIRT1;细胞凋亡;白内障

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中国实用眼科杂志

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期刊介绍： China Practical Ophthalmology was founded in May 1983. It is supervised by the National Health Commission of the People's Republic of China, sponsored by the Chinese Medical Association and China Medical University, and publicly distributed at home and abroad. It is a national-level excellent core academic journal of comprehensive ophthalmology and a series of journals of the Chinese Medical Association. China Practical Ophthalmology aims to guide and improve the theoretical level and actual clinical diagnosis and treatment ability of frontline ophthalmologists in my country. It is characterized by close integration with clinical practice, and timely publishes academic articles and scientific research results with high practical value to clinicians, so that readers can understand and use them, improve the theoretical level and diagnosis and treatment ability of ophthalmologists, help and support their innovative development, and is deeply welcomed and loved by ophthalmologists and readers.