Clinical College of Ophthalmology, Tianjin Medical University, Tianjin Eye Hospital, Tianjin, People's Republic of China.
Department of Ophthalmology, Beijing Shijitan Hospital, Capital Medical University, Beijing, People's Republic of China.
Invest Ophthalmol Vis Sci. 2015 Jan 27;56(2):993-1001. doi: 10.1167/iovs.14-15860.
MicroRNA-181a (miR-181a) is thought to be involved in posterior capsule opacification (PCO). This study investigated the role of miR-181a in the proliferation, migration, and epithelial-mesenchymal transition (EMT) of lens epithelial cells (LECs).
The expression of miR-181a was detected in human PCO-attached LECs and LECs obtained from patients with anterior polar cataracts by quantitative RT-PCR (qRT-PCR). The proliferation of SRA01/04 cells transfected with miR-181a mimics was analyzed by MTT assays and bromodeoxyuridine (BrdU)-incorporation assays. The migration of SRA01/04 cells was evaluated by wound-healing assays and Transwell migration. Luciferase reporter assays were used to validate the regulation of a putative target of miR-181a.
The expression of miR-181a is decreased in human PCO-attached LECs and LECs obtained from patients with anterior polar cataracts. A significant decrease in proliferation was observed in SRA01/04 cells transfected with miR-181a mimics. The overexpression of miR-181a inhibited the migration ability of LECs. Downregulation of fibronectin, Slug, and cyclooxygenase-2 (COX-2) expression and upregulation of E-cadherin expression were induced in human PCO-attached LECs transfected with miR-181a mimics and miR-181a-overexpressing LECs obtained from patients with anterior polar cataracts. Furthermore, luciferase assays using a reporter carrying a putative miR-181a target site in the 3' untranslated region of c-Met, Slug, and COX-2 revealed that miR-181a directly targets c-Met, Slug, and COX-2.
These data reveal that miR-181a can inhibit the proliferation, migration, and EMT of LECs and suggest that the restoration of miRNA-181a expression may be a potential novel therapeutic target for the prevention and treatment of posterior capsule opacification.
微小 RNA-181a（miR-181a）被认为参与后囊膜混浊（PCO）的形成。本研究旨在探讨 miR-181a 在晶状体上皮细胞（LECs）增殖、迁移和上皮-间充质转化（EMT）中的作用。
通过实时定量 RT-PCR（qRT-PCR）检测人 PCO 相关 LECs 和前极性白内障患者 LECs 中 miR-181a 的表达。通过 MTT 分析和溴脱氧尿苷（BrdU）掺入实验分析转染 miR-181a 模拟物的 SRA01/04 细胞的增殖。通过划痕愈合实验和 Transwell 迁移实验评估 SRA01/04 细胞的迁移。荧光素酶报告实验用于验证 miR-181a 的潜在靶标调控。
miR-181a 在人 PCO 相关 LECs 和前极性白内障患者 LECs 中的表达降低。转染 miR-181a 模拟物的 SRA01/04 细胞增殖显著减少。miR-181a 的过表达抑制了 LECs 的迁移能力。在转染 miR-181a 模拟物的人 PCO 相关 LECs 和前极性白内障患者中，miR-181a 过表达的 LECs 中纤维连接蛋白、Slug 和环氧化酶-2（COX-2）的表达下调，E-钙黏蛋白的表达上调。此外，使用携带 3'非翻译区中 miR-181a 靶位点的报告基因进行的荧光素酶实验表明，miR-181a 可直接靶向 c-Met、Slug 和 COX-2。
这些数据表明 miR-181a 可抑制 LECs 的增殖、迁移和 EMT，提示恢复 miRNA-181a 的表达可能是预防和治疗后囊膜混浊的潜在新治疗靶点。
