miR-449a对上皮后囊混浊晶状体上皮细胞增殖、迁移及间质转化的影响及机制  

Effects of miR-449a on proliferation,migration and epithelial mesenchymal transition of lens epithelial cells with posterior capsular opacification

作　　者：徐晓玮 田爱军 陈红 郝微 XU Xiaowei;TIAN Aijun;CHEN Hong;HAO Wei(The Eye Hospital of Hebei Province,Xingtai 054001,China)

机构地区：[1]河北省眼科医院,河北邢台054001

出　　处：《山东医药》2018年第37期45-48,共4页Shandong Medical Journal

摘　　要：目的探讨miR-449a对后囊混浊晶状体上皮细胞增殖、迁移及上皮间质转化的影响及机制。方法常规培养后囊混浊组织晶状体上皮细胞(PCO-LECs)及正常附着组织晶状体上皮细胞(normal-LECs),分别用qRTPCR法、酶联免疫吸附法检测细胞内miR-449a及上皮钙黏蛋白(E-cadherin)、纤黏蛋白(fibronectin)表达。向PCOLECs中转染miR-449a特异性的mimics(mimics组)及scrambled序列(miR-NC组),分别用qRT-PCR、酶联免疫吸附法检测细胞内miR-449a及相关蛋白质表达,用MTT法检测细胞增殖活力,用Transwell法检测细胞迁移能力,用双荧光素酶实验验证miR-449a与c-Met、c-Myc的靶向关系。结果与normal-LECs比较,PCO-LECs中miR-449a、E-cadherin低表达,fibronectin高表达(P均<0. 05)。与miR-NC组比较,mimics组miR-449a高表达(P <0. 05),细胞增殖活力、迁移能力低(P均<0. 05),E-cadherin高表达(P均<0. 05),fibronectin低表达(P均<0. 05)。双荧光素酶实验结果表明miR-449a具有潜在靶向抑制c-Met、c-Myc 3'UTR端的能力。结论 miR-449a可抑制后囊混浊晶状体上皮细胞增殖、迁移及上皮间质转化,其作用机制可能与靶向抑制c-Myc、c-Met表达有关。Objective To investigate the effects of miR-449a on the proliferation,migration and epithelial mesenchymal transition(EMT)of lens epithelial cells(LECs)with posterior capsular opacification(PCO)and its mechanism.Methods After collecting LECs from patients with or without PCO which were PCO-LECs and normal-LECs,we used QRT-PCR to assess miR-449a expression level and Western blotting to investigate the expression levels of EMT related proteins E-cadherin and fibronectin.We transfected miR-449a-specific mimics(mimics group)and scrambled sequences(miR-NC group)into PCO-LECs.Besides,we evaluated the influence of miR-449a on proliferation,migration and EMT.In addition,we used dual luciferase assay to verify the interaction between miR-449a and c-Met or c-Myc.Results Compared with the normal-LECs group,miR-449a,and E-cadherin expression was significantly down-regulated in the PCO-LECs group,but fibronectin was up-regulated(all P<0.05).Compared with the miR-NC group,mimics group showed over-expression of miR-449a(P<0.05),resulting in cell viability and migration ability attenuation,lower expression level of fibronectin or higher expression level of E-cadherin.In addition,dual luciferase assay suggested that miR-449a had a potential role in targeting c-Met and c-Myc via binding to their 3′UTR.Conclusion The miR-449a inhibits the proliferation,migration,and EMT of LECs by inhibiting the c-Myc and c-Met expression.

关 键 词：后囊混浊 白内障 晶状体上皮细胞 微小RNA-449a 

分 类 号：R779.66[医药卫生—眼科]