The E156K mutation in the CRYAA gene affects the epithelial–mesenchymal transition and migration of human lens epithelial cells

Zhennan Zhao; Jiahui Chen; Yongxiang Jiang; Yi Lu

Heliyon (Jan 2024)

The E156K mutation in the CRYAA gene affects the epithelial–mesenchymal transition and migration of human lens epithelial cells

Zhennan Zhao,
Jiahui Chen,
Yongxiang Jiang,
Yi Lu

Affiliations

Zhennan Zhao: Department of Ophthalmology and Vision Science, Eye Ear Nose and Throat Hospital of Fudan University, Shanghai, China; National Health Commission Key Laboratory of Myopia (Fudan University), Laboratory of Myopia, Chinese Academy of Medical Sciences, Shanghai, China; Key Laboratory of Visual Impairment and Restoration of Shanghai, Shanghai, China
Jiahui Chen: Department of Ophthalmology and Vision Science, Eye Ear Nose and Throat Hospital of Fudan University, Shanghai, China; National Health Commission Key Laboratory of Myopia (Fudan University), Laboratory of Myopia, Chinese Academy of Medical Sciences, Shanghai, China; Key Laboratory of Visual Impairment and Restoration of Shanghai, Shanghai, China
Yongxiang Jiang: Department of Ophthalmology and Vision Science, Eye Ear Nose and Throat Hospital of Fudan University, Shanghai, China; National Health Commission Key Laboratory of Myopia (Fudan University), Laboratory of Myopia, Chinese Academy of Medical Sciences, Shanghai, China; Key Laboratory of Visual Impairment and Restoration of Shanghai, Shanghai, China; Corresponding author. Eye Institute and Department of Ophthalmology, Eye & ENT Hospital, Fudan University, Shanghai, 200031, China.
Yi Lu: Department of Ophthalmology and Vision Science, Eye Ear Nose and Throat Hospital of Fudan University, Shanghai, China; National Health Commission Key Laboratory of Myopia (Fudan University), Laboratory of Myopia, Chinese Academy of Medical Sciences, Shanghai, China; Key Laboratory of Visual Impairment and Restoration of Shanghai, Shanghai, China; Corresponding author. Eye Institute and Department of Ophthalmology, Eye & ENT Hospital, Fudan University, Shanghai, 200031, China.

Journal volume & issue: Vol. 10, no. 1
p. e23690

Abstract

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Purpose: To investigated the biological effects of E156K-mutated αA-crystallin (CRYAA) in human lens epithelial cells (HLECs). Methods: FLAG-tagged, human, full-length, wild-type (WT), or E156K-mutated CRYAA was expressed in HLECs under CRYAA knockdown. CRYAA expression was determined by quantitative reverse transcription polymerase chain reaction and western blotting (WB). Rhodamine cytoskeleton staining was used to observe the changes in cell morphology following transfection with WT or E156K-mutated CRYAA plasmids. WB was performed to assess the expression of markers related to epithelial–mesenchymal transition (EMT) and migration. Results: Rhodamine cytoskeleton staining revealed changes in the morphology of cells transfected with E156K-mutated CRYAA and opposite responses occurred after treatment with a β-catenin inhibitor. Cells transfected with E156K-mutated CRYAA expressed remarkably higher levels of the mesenchymal biomarkers N-cadherin and vimentin but decreased levels of the epithelial biomarker E‐cadherin, whereas opposite trends were observed in cells treated with the β-catenin inhibitor, ICG001. The migratory capability of E156K-mutated CRYAA cells was significantly greater than that of WT cells (P < 0.001). This effect was accompanied by significantly increased expression levels of phosphorylated (p)-focal adhesion kinase (FAK) and p-Src. These changes were decreased significantly by treatment with FAK and Src inhibitors. Conclusion: E156K-mutated CRYAA induced EMT, in which the HLECs lost cell polarity, and acquired a mesenchymal phenotype with greater migratory capability. These biological effects may be associated with activation of the Wnt/β-Catenin and FAK/Src signaling pathways.

Published in Heliyon

ISSN: 2405-8440 (Online)
Publisher: Elsevier
Country of publisher: United Kingdom
LCC subjects: Science: Science (General); Social Sciences: Social sciences (General)
Website: https://www.cell.com/heliyon/home

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