Small Interfering RNA Targeted to ASPP2 Promotes Progression of Experimental Proliferative Vitreoretinopathy.
Small Interfering RNA Targeted to ASPP2 Promotes Progression of Experimental Proliferative Vitreoretinopathy.
复制标题
靶向 ASPP2 的小干扰 RNA 促进实验性增殖性玻璃体视网膜病变的进展
DOI：
10.1155/2016/7920631
复制
发表时间：
2016
影响因子：
4.6
通讯作者：
Li XX
中科院分区：
文献类型：
--
作者：
Chen XL;Bai YJ;Hu QR;Li SS;Huang LZ;Li XX
背景视网膜色素上皮（RPE）的上皮间质转化（EMT）在增生性玻璃体视网膜病变（PVR）的发生发展中起着重要作用。最近有报道p53的凋亡刺激蛋白（ASPP 2）参与EMT。然而，ASPP 2在PVR发病机制中的作用尚未确定。方法.应用免疫组化法检测PVR患者视网膜前膜中ASPP 2的表达。用ASPP 2-siRNA转染ARPE-19细胞，随后测量细胞毒性、增殖和迁移能力。免疫印迹法或流式细胞术检测EMT标志物及相关炎症和纤维化细胞因子。此外，通过玻璃体内注射用ASPP 2-siRNA转染的ARPE-19细胞诱导PVR大鼠模型，并相应地进行评价。结果免疫荧光分析显示ASPP 2在PVR膜中的表达较弱。ASPP 2敲低促进了RPE细胞的增殖和迁移，并增强了间充质标志物如α平滑肌肌动蛋白、纤连蛋白和ZEB 1的表达。同时，ASPP 2-siRNA增加EMT相关和炎性细胞因子，包括TGF-β、CTGF、VEGF、TNF-α和白细胞介素。PVR严重程度在ASPP 2-siRNA治疗的大鼠模型中更明显。结论. ASPP 2基因敲除可促进ARPE-19细胞的体外EMT，并可能通过炎症和纤维化细胞因子加剧体内实验性PVR的进展。
Background. Epithelial-mesenchymal transition (EMT) of retinal pigment epithelium (RPE) is vital in proliferative vitreoretinopathy (PVR) development. Apoptosis-stimulating proteins of p53 (ASPP2) have recently been reported to participate in EMT. However, the role of ASPP2 in PVR pathogenesis has not been identified. Methods. Immunohistochemistry was used to investigate the expression of ASPP2 in epiretinal membranes of PVR patients. ARPE-19 cells were transfected with ASPP2-siRNA, followed with measurement of cell cytotoxicity, proliferation, and migration ability. EMT markers and related inflammatory and fibrosis cytokines were measured by western blot or flow cytometry. Additionally, PVR rat models were induced by intravitreal injection of ARPE-19 cells transfected with ASPP2-siRNA and evaluated accordingly. Results. Immunofluorescence analysis revealed less intense expression of ASPP2 in PVR membranes. ASPP2 knockdown facilitated the proliferation and migration of RPE cells and enhanced the expression of mesenchymal markers such as alpha smooth muscle actin, fibronectin, and ZEB1. Meanwhile, ASPP2-siRNA increased EMT-related and inflammatory cytokines, including TGF-β, CTGF, VEGF, TNF-α, and interleukins. PVR severities were more pronounced in the rat models with ASPP2-siRNA treatment. Conclusions. ASPP2 knockdown promoted EMT of ARPE-19 cells in vitro and exacerbated the progression of experimental PVR in vivo, possibly via inflammatory and fibrosis cytokines.