Abstract
BackgroundRecent evidence has shown that long noncoding RNAs (lncRNAs) are involved in the process of epithelial-mesenchymal transition (EMT). However, little research has focused on the expression profile of lncRNAs during EMT in human lens epithelial cells (LECs) and their functions have not yet been described.MethodsDysregulated lncRNAs and mRNAs in normal human lens epithelial B-3(HLE B-3) cells and during transforming growth factor β2(TGF-β2)-induced EMT were analyzed via lncRNA microarray. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) Pathway analyses of differentially expressed mRNAs were performed to identify their functions and pathologic pathways. Six candidate lncRNAs were validated via quantitative real-time reverse transcription polymerase chain reaction(qRT-PCR) to confirm the microarray data.ResultsA total of 775 lncRNAs (325 up-regulated and 450 down-regulated) and 935 mRNAs (329 up-regulated and 606 down-regulated) were differentially expressed in HLE B-3 cells during TGF-β2-induced EMT compared to normal HLE B-3 cells. GO and KEGG Pathway analyses indicated the functions of differentially expressed mRNAs in the TGF-β2-induced EMT in HLE B-3 cells. qRT-PCR confirmed the trends indicated in microarray analysis for all 6 candidate lncRNAs.ConclusionOur study lays the foundation for future research in lncRNAs related to EMT in HLE B-3 cells and could provide new avenues for the prevention and treatment of posterior capsule opacification (PCO).
Highlights
Recent evidence has shown that long noncoding RNAs are involved in the process of epithelial-mesenchymal transition (EMT)
QRT-PCR/Western blot/Immunofluorescence of cells Phase-contrast micrographs showed, after stimulated with 10 ng/ml Transforming growth factor β2 (TGF-β2), cells were transformed from single polygonal to long and spindle-shaped (Fig. 1)
The long noncoding RNAs (lncRNAs) and mRNA expression patterns across the samples were distinguishable in the heat map generated by hierarchical clustering (Fig. 2c, d)
Summary
Recent evidence has shown that long noncoding RNAs (lncRNAs) are involved in the process of epithelial-mesenchymal transition (EMT). Little research has focused on the expression profile of lncRNAs during EMT in human lens epithelial cells (LECs) and their functions have not yet been described. Posterior capsule opacification (PCO) is the most common complication after cataract surgery [3]. The residual lens epithelial cells (LECs) in the anterior capsule after cataract surgery, over proliferate, and migrate to the posterior capsule, and epithelial-mesenchymal transition (EMT) takes place. This dynamic process is the main mechanism of PCO [4,5,6]. TGF-β2-induced EMT is a proper model for studying PCO [10]
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