Circ_0057452通过调节TGF-β2表达和吸附miR-145-5p，作为一种竞争性内源RNA在增生性瘢痕成纤维细胞增殖和VEGF表达中发挥作用。
Circ_0057452 functions as a ceRNA in hypertrophic scar fibroblast proliferation and VEGF expression by regulating TGF-β2 expression and adsorbing miR-145-5p.
作者信息
Qi Xiaoliu, Liu Yuxin, Yang Ming
机构信息
Department of Burn and Plastic Surgery, The Children's Hospital, Zhejiang University School of Medicine, National Clinical Research Center for Child Health and Disorders Hangzhou, Zhejiang Province, China.
出版信息
Am J Transl Res. 2021 Jun 15;13(6):6200-6210. eCollection 2021.
OBJECTIVE
To explore the mechanism by which circ_0057452/miR-145-5p/TGF-β2 axis regulates fibroblast proliferation as well as VEGF expression in hypertrophic scars (HS).
METHODS
The expression of circ_0057452, miR-145-5p and TGF-β2 in HS tissues and fibroblasts was measured by quantitative real-time Polymerase Chain Reaction (qRT-PCR). The targeting relations between circ_0057452 and miR-145-5p, miR-145-5p and TGF-β2 were identified using dual-luciferase reporter assay. The expression of circ_0057452, miR-145-5p and TGF-β2 in fibroblasts was interfered with and cells were grouped. In each group, changes in cell proliferation were detected using CCK8 assay, apoptosis was measured by flow cytometry, and VEGF secreted in cell culture supernatant was tested by ELISA kit.
RESULTS
Compared with normal tissues and fibroblasts, the expressions of circ_0057452 and TGF-β2 were increased and miR-145-5p decreased in HS tissues and cells (all P<0.05). Compared with the si-NC group, cell proliferation and VEGF expressions were decreased and the apoptotic rate increased in the si_circ_0057452 group (all P<0.05). Compared with the oe-NC group, cell proliferation and VEGF expression were increased and the apoptotic rate decreased in the oe-circ_0057452 group (all P<0.05). Compared with the oe-circ_0057452 + miR-NC group, the number of apoptotic cells was increased, and cell proliferation, as well as VEGF expression were decreased in the oe-circ_0057452 + miR-145-5p mimic group (all P<0.05). Compared with the miR-NC group, cell proliferation and VEGF expression were reduced and the apoptotic rate was increased in the miR-145-5p mimic group (all P<0.05). Compared with the miR-145-5p mimic + vector group, cell proliferation and VEGF expression were elevated, and apoptosis was inhibited in the miR-145-5p mimic + TGF-β2 group (all P<0.05).
CONCLUSION
circ_0057452 can competitively bind miR-145-5p to induce the expression of TGF-β2, and then promote the proliferation of HS fibroblasts and secretion of VEGF, which is expected to be effective in the treatment of HS.
目的
探讨环状RNA_0057452/微小RNA-145-5p/转化生长因子-β2（circ_0057452/miR-145-5p/TGF-β2）轴调控增生性瘢痕（HS）中成纤维细胞增殖及血管内皮生长因子（VEGF）表达的机制。
方法
采用定量实时聚合酶链反应（qRT-PCR）检测HS组织及成纤维细胞中circ_0057452、miR-145-5p和TGF-β2的表达。运用双荧光素酶报告基因检测法鉴定circ_0057452与miR-145-5p、miR-145-5p与TGF-β2之间的靶向关系。干扰成纤维细胞中circ_0057452、miR-145-5p和TGF-β2的表达并进行细胞分组。每组中，采用CCK8法检测细胞增殖变化，通过流式细胞术检测细胞凋亡情况，利用酶联免疫吸附测定（ELISA）试剂盒检测细胞培养上清液中分泌的VEGF。
结果
与正常组织及成纤维细胞相比，HS组织及细胞中circ_0057452和TGF-β2的表达升高，miR-145-5p的表达降低（均P<0.05）。与小干扰RNA阴性对照（si-NC）组相比，小干扰RNA circ_0057452（si_circ_0057452）组细胞增殖及VEGF表达降低，凋亡率升高（均P<0.05）。与过表达阴性对照（oe-NC）组相比，过表达circ_0057452（oe-circ_0057452）组细胞增殖及VEGF表达升高，凋亡率降低（均P<0.05）。与oe-circ_0057452+微小RNA阴性对照（miR-NC）组相比，oe-circ_0057452+miR-145-5p模拟物组凋亡细胞数量增加，细胞增殖及VEGF表达降低（均P<0.05）。与miR-NC组相比，miR-145-5p模拟物组细胞增殖及VEGF表达降低，凋亡率升高（均P<0.05）。与miR-145-5p模拟物+载体组相比，miR-145-5p模拟物+TGF-β2组细胞增殖及VEGF表达升高，凋亡受到抑制（均P<0.05）。
结论
circ_0057452可竞争性结合miR-145-5p以诱导TGF-β2表达，进而促进HS成纤维细胞增殖及VEGF分泌，有望用于HS的治疗。
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