微小RNA-30a在后发性白内障中对上皮间质转化的调控机制  

Mechanism of miR-30a regulates epithelial-mesenchymal transition in posterior capsule opacification

作　　者：马君择[1] 柏凌[1] 刘子瑶[1] 范雅稚[1] 权彦龙[1] Ma Jun-ze;Bai Ling;Liu Zi-yao;Fan Ya-zhi;Quan Yan-long(Department of Ophthalmology,Second Affiliated Hospital of Xi'an Jiaotong University,Xi'an 710004,China)

机构地区：[1]西安交通大学第二附属医院眼科,陕西西安710004

出　　处：《兰州大学学报（医学版）》2018年第4期16-22,共7页Journal of Lanzhou University（Medical Sciences）

基　　金：陕西省重点研究发展计划项目(2017SF-162).

摘　　要：目的探讨微小RNA(miR)-30a在后发性白内障(PCO)中对晶状体上皮细胞上皮间质转化(EMT)的作用及调控机制。方法通过实时荧光定量PCR检测43例PCO晶状体上皮组织和18例正常晶状体上皮组织中的miR-30a和EMT标志物E-cadherin、Fibronectin表达水平,并通过Pearson相关系数分析miR-30a与EMT标志物表达水平的相关性。检测过表达miR-30a后对人晶状体上皮细胞株SAR01/04增殖和侵袭能力的影响,以及对EMT相关蛋白的表达调控。利用miRNA靶基因预测软件预测miR-30a的靶基因,并通过荧光素酶报告基因进行靶基因和结合序列验证。在PCO和正常晶状体上皮原代细胞中过表达miR-30a后检测EMT相关蛋白的表达差异。结果miR-30a和E-cadherin在PCO晶状体上皮细胞中呈低表达水平,而Fibronectin呈高表达水平。miR-30a与E-cadherin呈显著正相关关系(r=0.754,P<0.001),而miR-30a与Fibronectin呈显著负相关关系(r=-0.857,P<0.001)。过表达miR-30a后能够降低人晶状体上皮细胞株SAR01/04的增殖和侵袭活力、上调E-cadherin表达、下调Fibronectin、Cox-2和Slug的表达。报告基因分析显示miR-30a能够通过结合到Slug基因的3’-UTR序列而下调Slug的表达。在PCO和正常晶状体上皮原代细胞中过表达miR-30a能上调E-cadherin表达、下调Fibronectin、Slug和Cox-2的表达。结论miR-30a通过调控Slug的表达来抑制人晶状体上皮细胞的EMT发生而减弱增殖和侵袭。Objective To investigate into the role and regulation mechanism of microRNA(miR)-30 a in epithelial to mesenchymal transition(EMT)of posterior capsular cataract(PCO).Methods The expression levels of miR-30 a and EMT markers(E-cadherin,Fibronectin)in 43 cases of PCO lens epithelium and 18 cases of normal lens epithelium were detected by real-time quantitative PCR.Pearson correlation coefficient was used to analyze the correlation between miR-30 a and EMT markers.The effect of overexpression of miR-30 a on the proliferation and invasion of human lens epithelial cell line SAR01/04,as well as the EMT-related protein expression were detected.The miR-30 a target gene was predicted by miRNA target gene prediction software,validated by the luciferase reporter assay.The overexpression of miR-30 a in PCO and normal lens epithelial primary cells was followed by detection of the differential expression of EMT-related proteins.Results miR30 a and E-cadherin were lowly expressed in PCO lens epithelial cells,while Fibronectin highly expressed.There was a significant positive correlation between miR-30 a and E-cadherin(r=0.754,P<0.001),but a negative correlation between miR-30 a and Fibronectin(r=-0.857,P<0.001).Overexpression of miR-30 a reduced the proliferation and invasion ability of human lens epithelial cell line SAR01/04,followed by E-cadherin up-regulation,and Fibronectin,Slug,and Cox-2 down-regulation.Reporter gene assay revealed that miR-30 was capable of down-regulating the Slug expression by binding to the 3’-UTR sequence of the Slug gene.Overexpression of miR-30 a in PCO and normal lens epithelial cells could up-regulate E-cadherin expression and down-regulate the expression of Fibronectin,Slug,and Cox-2.Conclusions miR-30 a attenuates proliferation and invasion by inhibiting EMT in human lens epithelial cells by down-regulating Slug expression.

关 键 词：后发性白内障 微小RNA-30a 上皮间质转化 调控 

分 类 号：R776.1[医药卫生—眼科]