Liu Lei, Xiao Wei
Department of Ophthalmology, Shengjing Hospital, China Medical University, NO.36 Sanhao Street, Shenyang City, Liaoning Province, 110004, China.
Department of Ophthalmology, Shenyang The Fourth Hospital of People, NO.20 Huanghe South Street, Shenyang City, Liaoning Province, 110031, China.
BMC Ophthalmol. 2017 Aug 1;17(1):135. doi: 10.1186/s12886-017-0532-1.
Posterior Capsular Opacification (PCO) is one of the most common complications of cataract surgery which can result in severe visual damage. Epithelial-Mesenchymal Transition (EMT) of lens epithelium cells (LEC) is the pathological basis of PCO. Recent research showed that hypoxia acted as an inducer of EMT through a Notch1/Snail1/E-cadherin pathway. However, it remains unclear whether the Notch1/Snail1/E-cadherin pathway is involved in PCO under hypoxia.
The morphology of SRA01/04 cells treating with Cobalt Chloride (CoCl) was observed and the markers of EMT and Notch1/Snail1/E-cadherin pathway were analyzed by Western blot and Immunocytochemistry assay. Transwell invasion assay and Wound healing assay were used to detected the effect of p3 × FLAG-CMV-7-NICD1 transfection on the SRA01/04 cells.
The SRA01/04 cells lost cell polarity and cell junction culturing with CoCl. The expression of Keratin, Hypoxia-inducible factor-1 alpha (HIF-1α), Notch1, Snail1were upregulated, on the other side, Fibronectin and E-cadherin were downregulated in hypoxia. Furthermore, the overexpression of Notch1 induced the expression of E-cadherin and increased the invasion and migration ability of SRA01/04 cells.
These results suggest that Notch1/Snail1/E-cadherin pathway facilitates the EMT through HIF-1α in SRA01/04 cells during hypoxia and promotes LEC motility.
后囊膜混浊（PCO）是白内障手术最常见的并发症之一，可导致严重的视力损害。晶状体上皮细胞（LEC）的上皮-间质转化（EMT）是PCO的病理基础。最近的研究表明，缺氧通过Notch1/Snail1/E-钙黏蛋白途径作为EMT的诱导剂。然而，缺氧条件下Notch1/Snail1/E-钙黏蛋白途径是否参与PCO仍不清楚。
观察用氯化钴（CoCl）处理的SRA01/04细胞的形态，并通过蛋白质免疫印迹和免疫细胞化学分析EMT和Notch1/Snail1/E-钙黏蛋白途径的标志物。采用Transwell侵袭实验和伤口愈合实验检测p3×FLAG-CMV-7-NICD1转染对SRA01/04细胞的影响。
用CoCl培养时，SRA01/04细胞失去细胞极性和细胞连接。缺氧时，角蛋白、缺氧诱导因子-1α（HIF-1α）、Notch1、Snail1的表达上调，而纤连蛋白和E-钙黏蛋白的表达下调。此外，Notch1的过表达诱导了E-钙黏蛋白的表达，并增加了SRA01/04细胞的侵袭和迁移能力。
这些结果表明，在缺氧条件下，Notch1/Snail1/E-钙黏蛋白途径通过HIF-1α促进SRA01/04细胞的EMT，并促进LEC的运动。
