Yin Ruoxi, Chen Xiaoya
Postgraduate School, Xuzhou Medical University, Xuzhou Eye Hospital, Xuzhou, Jiangsu 221000, P.R. China.
Department of Ophthalmology, Xuzhou No. 1 People's Hospital, Xuzhou Municipal Hospital Affiliated to Xuzhou Medical University, Xuzhou Eye Hospital, Xuzhou, Jiangsu 221000, P.R. China.
Exp Ther Med. 2019 Jul;18(1):647-653. doi: 10.3892/etm.2019.7584. Epub 2019 May 15.
Role of microRNA (miR)-144-3p in regulating the function of human trabecular meshwork cells (HTMCs) and fibronectin-1 (FN-1) was investigated. HTM cell lines were divided into five groups, of which four groups established oxidative stress HTMC models and one served as a control group. The four model groups were transfected with miR-144-3p independent sequence, inhibitory sequence, over-expression sequence, and the blank group received no transfection. In addition, 40 primary open angle glaucoma patients treated in Xuzhou No. 1 People's Hospital were included in the observation group, and 40 healthy individuals were enrolled as a normal group. RT-qPCR was used for the detection of miR-144-3p expression in serum and cells of patients and healthy people in each group, western blot analysis for FN-1 expression in cells, CCK-8 kit for cell proliferation, and Transwell for cell invasion. The expression of serum miR-144-3p in the observation group was significantly lower than that in the normal group (P<0.05). The cell optical density value in the over-expression group was significantly higher than that in the other groups (P<0.05), and in the inhibition group was significantly lower than that in the other groups (P<0.05). The number of cell-penetration in the over-expression group was significantly higher than that in the other groups (P<0.05). The expression of FN-1 protein in the over-expression group was significantly lower than that in the other four groups (P<0.05), and the expression in the inhibition group was significantly higher than that in the other four groups (P<0.05). The over-expression of miR-144-3p promotes proliferation and invasion of HTMCs by inhibiting the expression of FN-1 in inoxidative stress HTMCs, and is a potential target for glaucoma treatment.
研究了微小RNA(miR)-144-3p在调节人小梁网细胞(HTMCs)功能和纤连蛋白-1(FN-1)中的作用。HTM细胞系分为五组,其中四组建立氧化应激HTMC模型,一组作为对照组。四个模型组分别转染miR-144-3p独立序列、抑制序列、过表达序列,空白组不进行转染。此外,选取徐州市第一人民医院收治的40例原发性开角型青光眼患者作为观察组,选取40例健康个体作为正常组。采用RT-qPCR检测各组患者和健康人血清及细胞中miR-144-3p的表达,采用蛋白质免疫印迹法分析细胞中FN-1的表达,采用CCK-8试剂盒检测细胞增殖,采用Transwell检测细胞侵袭。观察组血清miR-144-3p表达明显低于正常组(P<0.05)。过表达组细胞光密度值明显高于其他组(P<0.05),抑制组明显低于其他组(P<0.05)。过表达组细胞穿膜数明显高于其他组(P<0.05)。过表达组FN-1蛋白表达明显低于其他四组(P<0.05),抑制组明显高于其他四组(P<0.05)。miR-144-3p的过表达通过抑制非氧化应激HTMCs中FN-1的表达促进HTMCs的增殖和侵袭,是青光眼治疗的潜在靶点。