Upregulation of EphA2 is associated with apoptosis in response to H2O2 and UV radiation-induced cataracts

被引：2

作者：

Zhao, Wei ^{[1
]}

Chen, Shuying ^{[1
]}

Lu, Bing ^{[1
]}

Wu, Di ^{[1
]}

Gu, Yuzhou ^{[1
]}

Hao, Shengjie ^{[1
]}

Sheng, Feiyin ^{[1
]}

Xu, Yili ^{[1
]}

Han, Yu ^{[1
]}

Chen, Rongrong ^{[1
]}

Zhou, Lei ^{[2
,3
]}

Fu, Qiuli ^{[1
]}

Yao, Ke ^{[1
]}

机构：

[1] Zhejiang Univ, Zhejiang Prov Engn Inst Eye Dis,Affiliated Hosp 2, Zhejiang Prov Clin Res Ctr Eye Dis,Eye Ctr, Zhejiang Prov Key Lab Ophthalmol,Sch Med, Hangzhou, Zhejiang, Peoples R China

[2] Hong Kong Polytech Univ, Res Ctr SHARP Vis RCSV, Sch Optometry, Dept Appl Biol & Chem Technol, Hong Kong, Peoples R China

[3] Ctr Eye & Vis Res CEVR, 17W Hong Kong Sci Pk, Hong Kong, Peoples R China

来源：

ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS | 2023年 / 747卷

基金：

中国国家自然科学基金;

关键词：

EphA2; Cataract; Oxidative stress; Apoptosis; Crispr/Cas9 genome editing; CONGENITAL CATARACT; IN-VIVO; LENS; EXPRESSION; EXPOSURE; GROWTH; IRRADIATION; GENERATION; MECHANISM; MUTATION;

D O I：

10.1016/j.abb.2023.109756

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

In this article, we examine the role of erythropoietin-producing hepatocellular receptor A2 (EphA2) in the apoptosis of lens epithelial cells (LECs) in H2O2 and UV radiation-induced cataracts. We treated SRA01/04 cells with H2O2 or ultraviolet (UV) radiation to create a cataract cell model. We constructed a cataract lens model by exposing mice to UV radiation. We used CCK8 assays, Annexin V-FITC analysis, and immunohistochemical staining to explore proliferation and apoptosis of the cataract model. Thereafter, we used quantitative real-time PCR (qPCR) analysis, Western blot assays, and immunofluorescence to determine gene and protein expression levels. We also employed Crispr/Cas9 gene editing to create an EphA2 knockout in SRA01/04 cells. Results: H2O2 or UV radiation induced SRA01/04 cells showed EphA2 gene upregulation. CCK8 and apoptosis assays showed that EphA2 over-expression (OE) reduced epithelial cell apoptosis, but knockout of EphA2 induced it in response to H2O2 and UV radiation, respectively. Mutation of the EphA2 protein kinase domain (c.2003G > A, p. G668D) had a limited effect on cell apoptosis. In vivo, the EphA2 protein level increased in the lenses of UV-treated mice. Our results showed that EphA2 was upregulated in SRA01/04 cells in response to H2O2 and UV radiation. Mutation of the EphA2 protein kinase domain (c.2003G > A, p. G668D) had a limited effect on H2O2 and UV radiation-induced cell apoptosis. We confirmed this change with an experiment on UV-treated mice. The present study established a novel association between EphA2 and LEC apoptosis.

引用

页数：10