Zhang Rui, Wei You-Heng, Zhao Chun-Yan, Song Hong-Yuan, Shen Ni, Cui Xiao, Gao Xin, Qi Zhong-Tian, Zhong Ming, Shen Wei
Department of Ophthalmology, Changhai Hospital, Second Military Medical University, Shanghai 200433, China.
State Key Laboratory of Genetic Engineering, Institute of Genetics, School of Life Sciences, Fudan University, Shanghai 200438, China.
Int J Ophthalmol. 2018 Jan 18;11(1):18-24. doi: 10.18240/ijo.2018.01.04. eCollection 2018.
To study the effect of discoidin I-like domaincontaining protein 3 (EDIL3) depletion on the proliferation and epithelial-mesenchymal transition (EMT) in human lens epithelial cells (LECs).
RNA interference was used to inhibit the expression of EDIL3 in human LECs . The morphology of cells was observed using an inverted microscope. Cell proliferation was assessed using EdU kit. Cell migration was investigated using Transwell chamber and EMT of LECs was assessed using confocal microscope and Western blotting. The transforming growth factor β (TGFβ) pathway was investigated using Western blotting.
The data showed that silencing EDIL3 expression changed LECs morphology and suppressed LECs proliferation (<0.05) and migration (<0.01). Furthermore, the result of Western blotting showed that EDIL3 depletion reduced the expression of α-smooth muscle actin (α-SMA) (<0.001) and vimentin (<0.01), while increased the expression of E-cadherin (<0.001). EDIL3 depletion could suppress the phosphorylation of Smad2 (<0.01) and Smad3 (<0.01) and the activation of exracellular signal regulated kinase (ERK) (<0.05).
The findings indicate that EDIL3 might participate in the proliferation and EMT in LECs TGFβ pathway and may be a potential therapeutic target for the treatment of posterior capsule opacification.
研究含盘状结构域蛋白3（EDIL3）缺失对人晶状体上皮细胞（LECs）增殖及上皮-间质转化（EMT）的影响。
采用RNA干扰抑制人LECs中EDIL3的表达。用倒置显微镜观察细胞形态。使用EdU试剂盒评估细胞增殖。使用Transwell小室研究细胞迁移，并用共聚焦显微镜和蛋白质免疫印迹法评估LECs的EMT。用蛋白质免疫印迹法研究转化生长因子β（TGFβ）信号通路。
数据显示，沉默EDIL3表达改变了LECs的形态，抑制了LECs的增殖（<0.05）和迁移（<0.01）。此外，蛋白质免疫印迹结果显示，EDIL3缺失降低了α-平滑肌肌动蛋白（α-SMA）（<0.001）和波形蛋白（<0.01）的表达，同时增加了E-钙黏蛋白的表达（<0.001）。EDIL3缺失可抑制Smad2（<0.01）和Smad3（<0.01）的磷酸化以及细胞外信号调节激酶（ERK）的激活（<0.05）。
研究结果表明，EDIL3可能通过TGFβ信号通路参与LECs的增殖和EMT，可能是治疗后囊膜混浊的潜在治疗靶点。
