摘要
目的探究miRNA-21通过作用于Erk信号通路关键蛋白Erk的表达从而对视网膜色素上皮细胞的增殖侵袭和凋亡能力的影响。方法通过转染miR-21 mimics、miR-21 inhibitors以及对照物实现人视网膜色素上皮细胞系ARPE-9中miR-21表达的改变,Q-PCR法检测miR-21,Erk基因的表达,用Western blot法检测Erk蛋白的表达水平,MTS法测定ARPE-9细胞的增殖,transwell检测细胞的侵袭,Annexin V-FITC/PI双染法检测细胞凋亡。结果与阴性对照组相比,转染miR-21 mimics/miR-21 inhibitors分别引起细胞中miR-21,Erk表达的升高/降低,增殖能力上调/下调,侵袭能力上调/下调,凋亡水平下调/上调。结论ARPE-9细胞中miRNA-21表达的改变可以引起Erk表达水平的改变,同时引起细胞的增殖侵袭和凋亡水平的改变,此研究对视网膜病变以及后续的机制研究提供了参考。
Objective To explore the effect of miRNA-21 on the proliferation,invasion and apoptosis of retinal pigment epithelial(RPE)cells by acting on the expression of Erk,a key protein of Erk signaling pathway.Methods Transfection of miR-21 mimics,miR-21 inhibitors,and control substances were used to change the expression of miR-21 in the human RPE cell line ARPE-9.Q-PCR method was used to detect the expression of miR-21 and Erk genes.Western blot method was used to detect the expression level of Erk protein,MTS method was used to detect the proliferation of ARPE-9 cells,transwell was used to detect cell invasion,and the Annexin V-FITC/PI double staining method was used to detect cell apoptosis.Results Compared with the negative control group,transfection of miR-21 mimics/miR-21 inhibitors caused increased/decreased expression of miR-21 and Erk,increased/decreased proliferation ability,increased/decreased invasion ability,and decreased/increased apoptosis level,respectively.Conclusions The changes of miRNA-21 expression in ARPE-9 cells can cause changes in the expression level of Erk,as well as changes in the levels of cell proliferation,invasion and apoptosis.This study provides a reference for the retinopathy and subsequent mechanism studies.
作者
冯婷婷
何广辉
董蒙
武斌
梁泽玉
陈松
Feng Tingting;Chen Song;He Guanghui;Liang Zeyu;Dong Meng;Wu Bin(Department of Tianjin Eye Hospital, Clinical College of Ophthalmology of Tianjin Medical University, Tianjin Key Lab of Ophthalmology and Visual Science, Tianjin Eye Institute, Tianjin 300020, China)
出处
《临床眼科杂志》
2021年第2期165-169,共5页
Journal of Clinical Ophthalmology
基金
天津市科学技术委员会基金资助项目(14JCYBJC27400)。