Original Article
miR-211 inhibitor up-regulates the progression of age-related cataract by targeting Notch22-mediated expression of β-crystallin B2
Shi Zhan, Wang Chao, Wang Feng
Published 2019-09-05
Cite as Int J Immunol, 2019,42(5): 449-455. DOI: 10.3760/cma.j.issn.1673-4394.2019.05.001
Abstract
ObjectiveTo investigate the role and its potential mechanism of microRNA-211(miR-211) and β-crystalin B2 in aging-related cataract.
MethodsCCK-8 and flow cytometry were used to detect the cell growth of HLEC-3 cell after treated with different concentrations of hydrogen peroxide or different expressions of miRNA-211.Bioinformatics method was used to predict the potential binding sites of miRNA-211 to Notch2.Westren blot and quantitative reverse transcription polymerase chain reaction(qRT-PCR) were used to detect the expression of Nocth2 protein and Mrna in HLEC-3 cells after treatment with miR-211.Luciferase reporter was used to confirm the direct target gene of miR-211.Western blot was used to detect the changes of beta-crystallin B2 protein after single or co-expression of miRNA-211+ Notch2 or low expression of Notch2.
ResultsHydrogen peroxide (H2O2) could significantly reduce the cell viability of HLCE-3 cells(t=3.67, P<0.05). Down-regulation of miRNA-211 can significantly inhibit the apoptosis of HLEC-B3 induced by H2O2(t=4.07, P<0.05). However, up-regulation of miR-211-211 can significantly increase the expression of oxidizing factors(t=4.21, P<0.05). Luciferase and Western blot studies showed that up-regulation of miRNA-211 targeted inhibition of Notch2 expression(t values were 4.02 and 3.90, both P values <0.05). Similarly, down-regulation of miR-211 could revised the expression of Notch2(t=4.04, P<0.05). In addition to Notch2, miR-211 inhibitor also enhanced β-crystallin B2 expression.Beta-crystallin B2 expression induced by miR-211 inhibitor could be significantly inhibited after Notch2 expression were silenced (t values were 4.20, 4.19 and 4.15 respectively, all P values <0.05).
ConclusionsThese data demonstrated that miR-211 inhibitor increased β-crystallin B2 expression via up-regulating Notch2, which indicated that miR-211 inhibitor might be a promising candidate to prevent progression of age-related cataract.
Key words:
microRNA-211; Age-related cataract; Notch21; β-crystallin B2
Contributor Information
Shi Zhan
Department of Ophthalmology, the First Affiliated Hospital, Harbin Medical University, Harbin 150001, China
Wang Chao
Department of Anesthesia, the First Affiliated Hospital, Harbin Medical University, Harbin 150001, China
Wang Feng
Department of Ophthalmology, the First Affiliated Hospital, Harbin Medical University, Harbin 150001, China