June 2017
Volume 58, Issue 8
Open Access
ARVO Annual Meeting Abstract  |   June 2017
Long non-coding RNA H19 regulates human lens epithelial cells function: implication in the pathogenesis of age-related nuclear cataract
Author Affiliations & Notes
  • Xin Liu
    Ophthalmology, Eye and ENT hospital of Fudan University, Shanghai, China
  • Chang Liu
    Ophthalmology, Eye and ENT hospital of Fudan University, Shanghai, China
  • Kun Shan
    Ophthalmology, Eye and ENT hospital of Fudan University, Shanghai, China
  • Shujie Zhang
    Ophthalmology, Eye and ENT hospital of Fudan University, Shanghai, China
  • Yi Lu
    Ophthalmology, Eye and ENT hospital of Fudan University, Shanghai, China
  • Biao Yan
    Ophthalmology, Eye and ENT hospital of Fudan University, Shanghai, China
  • Yi Luo
    Ophthalmology, Eye and ENT hospital of Fudan University, Shanghai, China
  • Footnotes
    Commercial Relationships   Xin Liu, None; Chang Liu, None; Kun Shan, None; Shujie Zhang, None; Yi Lu, None; Biao Yan, None; Yi Luo, None
  • Footnotes
    Support  None
Investigative Ophthalmology & Visual Science June 2017, Vol.58, 2040. doi:
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    • Get Citation

      Xin Liu, Chang Liu, Kun Shan, Shujie Zhang, Yi Lu, Biao Yan, Yi Luo; Long non-coding RNA H19 regulates human lens epithelial cells function: implication in the pathogenesis of age-related nuclear cataract. Invest. Ophthalmol. Vis. Sci. 2017;58(8):2040.

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      © ARVO (1962-2015); The Authors (2016-present)

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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.

 

(A-C)Differential lncRNAexpression between transparent and nuclear ARC lenses samples.
(D-E)LncRNA H19 expression levels positively correlates with nuclear ARC grade.
(F-M)Effects of H19 knockdown on HLECs function.H19 expression levels,cell viability, apoptotic cells, mitochondrial membrane potentials, HLECs proliferation and migration were detected.

(A-C)Differential lncRNAexpression between transparent and nuclear ARC lenses samples.
(D-E)LncRNA H19 expression levels positively correlates with nuclear ARC grade.
(F-M)Effects of H19 knockdown on HLECs function.H19 expression levels,cell viability, apoptotic cells, mitochondrial membrane potentials, HLECs proliferation and migration were detected.

 

(A-F)H19/miR675-5p downregulates the expression of CRYAA.
(G-M)miR675-5p is involved in the regulation of HLEC function. Cell viability, apoptotic cells, mitochondrial membrane potentials, HLECs proliferation and migration were detected.

(A-F)H19/miR675-5p downregulates the expression of CRYAA.
(G-M)miR675-5p is involved in the regulation of HLEC function. Cell viability, apoptotic cells, mitochondrial membrane potentials, HLECs proliferation and migration were detected.

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