Chen You, Zhao Tong, Han Mengyu, Chen Yi
Department of Ophthalmology, China-Japan Friendship Hospital Beijing 100029, China.
Am J Transl Res. 2024 Feb 15;16(2):446-457. doi: 10.62347/BXFG4038. eCollection 2024.
Cataract causes the greatest number of blindnesses worldwide. This study aims to investigate the role of miR-143 in lens epithelial cells.
Clustering analysis was conducted to systematically compare miRNA expression levels across cataract and myopia. The levels of miR-143 and Bromodomain containing 2 (BRD2) were determined using real-time quantitative PCR (RT-qPCR) assay in lens epithelial cells. Transwell and wound healing assays were conducted to detect cell invasive and migratory abilities. The regulation relationship between MiR-143 and BRD2 was assessed using dual-luciferase reporter gene assays. BRD2 was knocked down using siRNA-BRD2, and siRNA-BRD2, and miR-143 inhibitors were transfected into cells with lipofectamine 2000.
Through retrieving five databases, 2690 miRNAs were selected. Volcano plot results demonstrated that 200 miRNAs were differentially expressed between cataract and myopia, in which 152 miRNAs were upregulated and 48 miRNAs downregulated in myopia compared with cataract. MiR-143 was upregulated in cataract compared with myopia (P<0.05). MiR-143 inhibitor suppressed the proliferation, invasion and migration of lens epithelial cells (all P<0.05). Luciferase reporter assays confirmed that BRD2 was a miR-143 target gene in SRA01/04 cells. Knockdown of BRD2 promoted SRA01/04 cell proliferation, invasion and migration (all P<0.05). In addition, silencing of BRD2 partially reversed the functions of miR-143 inhibitor on proliferation, invasion and migration (all P<0.05).
MiR-143 suppresses lens epithelial cell proliferation, invasion and migration by regulating BRD2, which may support a novel therapeutic strategy for cataract patients.
白内障是全球导致失明人数最多的病因。本研究旨在探讨miR-143在晶状体上皮细胞中的作用。
进行聚类分析以系统比较白内障和近视患者的miRNA表达水平。采用实时定量PCR(RT-qPCR)检测晶状体上皮细胞中miR-143和含溴结构域2(BRD2)的水平。通过Transwell实验和伤口愈合实验检测细胞侵袭和迁移能力。利用双荧光素酶报告基因实验评估MiR-143与BRD2之间的调控关系。使用siRNA-BRD2敲低BRD2,并将siRNA-BRD2和miR-143抑制剂用脂质体2000转染到细胞中。
通过检索五个数据库,筛选出2690个miRNA。火山图结果显示,白内障和近视患者之间有200个miRNA差异表达,其中与白内障相比,近视患者中有152个miRNA上调,48个miRNA下调。与近视相比,白内障患者中miR-143上调(P<0.05)。miR-143抑制剂抑制了晶状体上皮细胞的增殖、侵袭和迁移(均P<0.05)。荧光素酶报告实验证实BRD2是SRA01/04细胞中miR-143的靶基因。敲低BRD2促进了SRA01/04细胞的增殖、侵袭和迁移(均P<0.05)。此外,沉默BRD2部分逆转了miR-143抑制剂对增殖、侵袭和迁移的作用(均P<0.05)。
MiR-143通过调控BRD2抑制晶状体上皮细胞的增殖、侵袭和迁移,这可能为白内障患者提供一种新的治疗策略。