Abstract
Purpose :
Age-related cataract (ARC) remains the leading cause of visual impairment among elder population. Long non-coding RNAs (lncRNAs) have emerged as potential regulators in cataractgenesis. However, the role of lncRNAs in nuclear ARC, a subtype of ARC, remains to be further elucidated.
Methods :
LncRNA sequencing was performed to identify differentially expressed lncRNAs between the capsules of transparent and nuclear ARC lenses. Expression validation was confirmed by qRT-PCR. MTT assay, Calcein-AM and propidium iodide double staining, rhodamine 123 and Hoechst double staining, EdU and transwell assay were used to determine the role of H19 or miR-675 in the viability, apoptosis, proliferation and migration of human lens epithelial cells (HLECs). Bioinformatics and luciferase reporter assays were used to identify the binding target of miR-675.
Results :
Sixty-three lncRNAs are differentially expressed between the capsules of transparent and nuclear ARC lenses. One top abundantly expressed lncRNA, H19, is significantly up-regulated in the nuclear ARC lens capsules and positively associated with nuclear ARC grade. H19 knockdown accelerates apoptosis development and reduces the proliferation and migration of HLECs upon oxidative stress. H19 is the precursor of miR-675 and reduction of H19 inhibits miR-675 expression. miR-675 regulates CRYAA expression by directly targeting the binding site within the 3’UTR. Moreover, miR-675 increases the proliferation and migration, and decreases the apoptosis of HLECs upon oxidative stress.
Conclusions :
H19 regulates human lens epithelial cells function through miR-675-mediated CRYAA expression in the pathogenesis of nuclear ARC. This study would provide a novel insight into the pathogenesis of nuclear ARC.
This is an abstract that was submitted for the 2017 ARVO Annual Meeting, held in Baltimore, MD, May 7-11, 2017.