Knockdown of lncRNA XIST prevents the epithelial-mesenchymal transition of TGF-β2-induced human lens epithelial cells via miR-124/Slug axis

XUE JIANG,HONG ZHANG

doi:10.32604/biocell.2022.016167

Abstract

Posterior capsular opacification (PCO) is linked to the pathological process of lens epithelial cells, which includes proliferation, migration, and epithelial-mesenchymal transition (EMT). Our goal was to investigate whether long noncoding RNA (lncRNA) XIST contributes to EMT via targeting miR-124/Slug axis in TGF-β2-induced SRA01/04 cells. EMT was induced by stimulating SRA01/04 cells with 10 ng/mL TGF-β2 for 24 h, and PCO microenvironment was simulated. The expressions levels of lncRNA XIST, miR-124, and Slug were measured by real-time polymerase chain reaction (RT-PCR) and western blot. The role and mechanism of lncRNA XIST in promoting EMT of TGF-β2-treated SRA01/04 cells were investigated by using cell transfection, cell counting kit-8 (CCK-8), immunofluorescence staining, transwell assay, wound-healing assay, RT-PCR, western blot and dual-luciferase reporter assay. The expression of Slug and lncRNA XIST was markedly increased, while miR-124 was downregulated in TGF-β2-treated SRA01/04 cells compared with the control group. Knockdown of lncRNA XIST reduced EMT, migration, and cell viability in TGF-β2-induced SRA01/04 cells; moreover, it adversely modulated miR-124 and adjusted the expression of Slug in SRA01/04 cells, while these effects were diminished by co-transfection with AMO-miR-124. Our data demonstrated that lncRNA XIST functioned as a competitive endogenous RNA (ceRNA) of miR-124 to modulate the expression level of Slug, thereby modulating EMT, migration, and cell viability in SRA01/04 cells. In conclusion, lncRNA XIST has a crucial role in promoting TGF-β2-induced EMT via modulating the miR-124/Slug axis in SRA01/04 cells, and it may provide a novel therapeutic option for PCO treatment.

Highlights

Posterior capsular opacification (PCO) is one of the most common complications in recent cataract surgeries; it is known as “secondary cataract” (Awasthi et al, 2009)
The expression levels of Slug and long noncoding RNA (lncRNA) X inactive specific transcript (XIST) were increased, whereas miR-124 was decreased in Transforming growth factor-β2 (TGF-β2)-induced in SRA01/04 cells To explore whether epithelial– mesenchymal transition (EMT) takes part in the pathogenesis of PCO and to determine the expression of various substances in the EMT cell model, we used different concentrations (0, 1, 5, 10 ng/mL) of TGF-β2 for treating SRA01/04 cells for 0, 6, 12, and 24 h to establish the EMT cell model in vitro
It mainly develops due to cell viability, migration, and EMT of residual human lens epithelial cells (HLECs) postoperatively (Li et al, 2019) ; EMT likely plays a core role in the PCO formation (Dong, 2019a), but the underlying mechanisms of molecular regulation in the development of PCO remain unclear

Summary

Introduction

Posterior capsular opacification (PCO) is one of the most common complications in recent cataract surgeries; it is known as “secondary cataract” (Awasthi et al, 2009). Within 2–5 years after the operation, 20%–40% of the patients experience vision decline due to PCO (Awasthi et al, 2009; Nibourg et al, 2015). The only effective treatment for PCO is Nd: YAG laser posterior capsulotomy, but the risk of complications still exists (Zhang and Xie, 2020). PCO mainly results from the pathological process of residual lens epithelial cells after cataract surgery and includes epithelial– mesenchymal transition (EMT), proliferation, and migration. EMT is the primary causal factor for PCO development (Dong, 2019a; Zhang et al, 2017).

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