Department of Ophthalmology, Kaohsiung Veterans General Hospital, Kaohsiung, Taiwan.
School of Medicine, National Yang-Ming University, Taipei, Taiwan.
PLoS One. 2019 Mar 14;14(3):e0213932. doi: 10.1371/journal.pone.0213932. eCollection 2019.
Age-related macular degeneration (AMD) is an ocular disease with retinal degeneration. Retinal pigment epithelium (RPE) degeneration is mainly caused by long-term oxidative stress. Kinase activity could be either protective or detrimental to cells during oxidative stress; however, few reports have described the role of kinases in oxidative stress. In this study, high-throughput screening of kinome siRNA library revealed that erb-b2 receptor tyrosine-protein kinase 2 (ERBB2) knockdown reduced reactive oxygen species (ROS) production in ARPE-19 cells during oxidative stress. Silencing ERBB2 caused an elevation in microtubule associated protein light chain C3-II (MAP1LC3B-II/I) conversion and sequesterone (SQSTM)1 protein level. ERBB2 deprivation largely caused an increase in autophagy-regulating protease (ATG4B) expression, a protease that negatively recycles MAP1LC3-II at the fusion step between the autophagosome and lysosome, suggesting ERBB2 might modulate ATG4B for autophagy induction in oxidative stress-stimulated ARPE-19 cells. ERBB2 knockdown also caused an accumulation of nuclear factor erythroid 2-related factor 2 (NRF2) and enhanced its transcriptional activity. In addition, ERBB2 ablation or treatment with autophagy inhibitors reduced oxidative-induced cytotoxic effects in ARPE-19 cells. Furthermore, ERBB2 silencing had little or no additive effects in ATG5/7-deficient cells. Taken together, our results suggest that ERBB2 may play an important role in modulating autophagic RPE cell death during oxidative stress, and ERBB2 may be a potential target in AMD prevention.
年龄相关性黄斑变性 (AMD) 是一种视网膜变性的眼部疾病。视网膜色素上皮 (RPE) 变性主要是由长期氧化应激引起的。激酶活性在氧化应激过程中对细胞可能具有保护作用,也可能具有损害作用;然而,很少有报道描述激酶在氧化应激中的作用。在这项研究中,通过高通量筛选激酶组 siRNA 文库发现,在氧化应激期间,表皮生长因子受体酪氨酸蛋白激酶 2 (ERBB2) 的敲低可减少 ARPE-19 细胞中活性氧 (ROS) 的产生。沉默 ERBB2 导致微管相关蛋白轻链 C3-II (MAP1LC3B-II/I) 转化率和自噬相关蛋白 SQSTM1 水平升高。ERBB2 剥夺在很大程度上导致自噬调节蛋白酶 (ATG4B) 表达增加,ATG4B 是一种在自噬体与溶酶体融合步骤中负回收 MAP1LC3-II 的蛋白酶,表明 ERBB2 可能通过调节 ATG4B 来诱导氧化应激刺激的 ARPE-19 细胞中的自噬。ERBB2 敲低也导致核因子红细胞 2 相关因子 2 (NRF2) 的积累并增强其转录活性。此外,ERBB2 缺失或自噬抑制剂处理可减少 ARPE-19 细胞中氧化诱导的细胞毒性作用。此外,在 ATG5/7 缺陷细胞中,ERBB2 沉默几乎没有或没有相加作用。总之,我们的结果表明,ERBB2 可能在调节氧化应激期间 RPE 细胞自噬性死亡中发挥重要作用,ERBB2 可能是 AMD 预防的潜在靶点。