Wu Hong-Lian, Shao Yan, Chen Zhen-Na, Zhang Hui, Zhang Xiao-Min, Li Xiao-Rong
Tianjin Key Laboratory of Retinal Functions and Diseases; Tianjin International Joint Research and Development Centre of Ophthalmology and Vision Science; Eye Institute and School of Optometry, Tianjin Medical University Eye Hospital, Tianjin 300384, China.
Department of Ophthalmology, Affiliated Hospital of Weifang Medical University, Weifang 261031, Shandong Province, China.
Int J Ophthalmol. 2022 Jun 18;15(6):894-904. doi: 10.18240/ijo.2022.06.06. eCollection 2022.
To evaluate the effect of miRNA-451 on rhesus macaque choroid-retinal endothelial (RF/6A) cell function and proteome profile.
The RF/6A cells were transfected with miRNA-451 mimic and inhibitor. The role of miRNA-451 on proliferation ability was evaluated by CCK-8 assay. Furthermore, iTRAQ quantitative proteomic analysis was applied to comprehensively illuminate the change of cellular proteins and biological function between different groups.
In miRNA-451 overexpression group, cell proliferation of RF/6A decreased both at 24h and 48h; while in miRNA-451 inhibition group, on the contrary, RF/6A cell proliferation was increased at 48h. Based on iTRAQ quantitative proteomic analysis, 23 differentially expressed proteins (DEPs) were detected in the comparison of miRNA-451 mimic and mimic control-transfected RF/6A cells, and 30 DEPs were identified in the comparison of RF/6A cells transfected with miRNA-451 inhibitor and inhibitor control. DEPs such as GORASP2, KRT1, SLC7A2, RIC8A, DDX42, CAP1, PCBP2 might be closely related to the inhibitory effect of miRNA-451 on RF/6A cell proliferation, while PCYT1A, MGAT1, TUBB, MCU, SIL1, BID, MSH6 might account for the positive effect of miRNA-451 inhibitor on RF/6A cell growth. PTPN1, as the only protein exhibiting an opposite trend between miRNA-451 mimic and inhibitor-transfected cells, was most likely accountable for the inhibition of miRNA-451 mimic on RF/6A cell growth, and the promotion of miRNA-451 inhibitor on RF/6A cell proliferation.
miRNA-451 overexpression can suppress the growth of RF/6A cells while knockdown of miRNA-451 can promote RF/6A cell viability. Among all DEPs, increased PTPN1 is most likely to account for the negative regulation of miRNA-451 on RF/6A proliferation. miRNA-451 can be a protective factor for neovascular disease of fundus regulating choroid retinal endothelial cell function.
评估miRNA - 451对恒河猴脉络膜视网膜内皮（RF/6A）细胞功能和蛋白质组图谱的影响。
用miRNA - 451模拟物和抑制剂转染RF/6A细胞。通过CCK - 8法评估miRNA - 451对增殖能力的作用。此外，应用iTRAQ定量蛋白质组分析全面阐明不同组之间细胞蛋白质和生物学功能的变化。
在miRNA - 451过表达组中，RF/6A细胞在24小时和48小时的增殖均下降；而在miRNA - 451抑制组中，相反，RF/6A细胞在48小时时增殖增加。基于iTRAQ定量蛋白质组分析，在miRNA - 451模拟物与模拟对照转染的RF/6A细胞比较中检测到23种差异表达蛋白（DEP），在miRNA - 451抑制剂与抑制剂对照转染的RF/6A细胞比较中鉴定出30种DEP。诸如GORASP2、KRT1、SLC7A2、RIC8A、DDX42、CAP1、PCBP2等DEP可能与miRNA - 451对RF/6A细胞增殖的抑制作用密切相关，而PCYT1A、MGAT1、TUBB、MCU、SIL1、BID、MSH6可能解释了miRNA - 451抑制剂对RF/6A细胞生长的积极作用。PTPN1作为在miRNA - 451模拟物和抑制剂转染细胞之间呈现相反趋势的唯一蛋白质，最有可能是miRNA - 451模拟物对RF/6A细胞生长抑制以及miRNA - 451抑制剂对RF/6A细胞增殖促进的原因。
miRNA - 451过表达可抑制RF/6A细胞生长，而敲低miRNA - 451可促进RF/6A细胞活力。在所有DEP中，PTPN1增加最有可能是miRNA - 451对RF/6A增殖负调控的原因。miRNA - 451可能是调节脉络膜视网膜内皮细胞功能的眼底新生血管疾病的保护因子。
