Department of Plastic Surgery, the First Affiliated Hospital of Zhengzhou University, Zhengzhou, China.
Cell Cycle. 2022 Dec;21(23):2471-2483. doi: 10.1080/15384101.2022.2102796. Epub 2022 Jul 25.
Increasing evidence has shown that circular RNAs (circRNAs) play critical roles in various diseases, including keloid. The purpose of this study was to investigate the role of circ_0057452 and related action mechanisms during the development of keloid. The expression levels of circ_0057452, microRNA-7-5p (miR-7-5p) and GRB2 associated binding protein 1 (GAB1) mRNA were determined by quantitative real-time PCR (qRT-PCR). Cell proliferation was evaluated using methylthiazolyldiphenyl-tetrazolium bromide (MTT) and 5-Ethynyl-2'-deoxyuridine (Edu) assays. Flow cytometry analysis was utilized to determine cell cycle distribution and cell apoptosis. Western blot assay was used to measure apoptosis-related, collagen synthesis-related, and GAB1 protein levels. Cell migration and invasion were detected by wound healing assay and transwell assay. The interaction between miR-7-5p and circ_0057452 or GAB1 was confirmed by dual-luciferase reporter, RNA pull-down, and RNA Immunoprecipitation (RIP) assays. Circ_0057452 and GAB1 were upregulated in keloid tissues and keloid fibroblasts (KFs), while miR-7-5p was downregulated. Circ_0057452 knockdown or miR-7-5p overexpression inhibited the proliferation, migration, invasion, and collagen synthesis and induced cell cycle arrest and apoptosis of KFs. MiR-7-5p was targeted by circ_0057452, and its inhibition overturned the effects of circ_0057452 knockdown. In addition, GAB1 was a target of miR-7-5p, and GAB1 upregulation abolished the role of miR-7-5p overexpression and circ_0057452 knockdown in KFs. Circ_0057452 regulated the expression of GAB1 by adsorbing miR-7-5p in KFs. Circ_0057452 knockdown suppressed keloid development by regulating miR-7-5p/GAB1 axis, which might provide a promising therapeutic target for keloid.
越来越多的证据表明，环状 RNA（circRNA）在包括瘢痕疙瘩在内的各种疾病中发挥着关键作用。本研究旨在探讨 circ_0057452 在瘢痕疙瘩发展过程中的作用及其相关作用机制。通过实时定量 PCR（qRT-PCR）测定 circ_0057452、microRNA-7-5p（miR-7-5p）和 GRB2 相关结合蛋白 1（GAB1）mRNA 的表达水平。通过甲基噻唑基二苯基四唑溴盐（MTT）和 5-乙炔基-2'-脱氧尿苷（Edu）测定法评估细胞增殖。通过流式细胞术分析测定细胞周期分布和细胞凋亡。Western blot 检测法用于测量凋亡相关、胶原合成相关和 GAB1 蛋白水平。通过划痕愈合试验和 Transwell 试验检测细胞迁移和侵袭。通过双荧光素酶报告、RNA 下拉和 RNA 免疫沉淀（RIP）试验证实 miR-7-5p 与 circ_0057452 或 GAB1 之间的相互作用。circ_0057452 和 GAB1 在瘢痕疙瘩组织和瘢痕疙瘩成纤维细胞（KFs）中上调，而 miR-7-5p 下调。circ_0057452 敲低或 miR-7-5p 过表达抑制 KFs 的增殖、迁移、侵袭和胶原合成，并诱导细胞周期停滞和凋亡。circ_0057452 靶向 miR-7-5p，其抑制作用逆转了 circ_0057452 敲低的作用。此外，GAB1 是 miR-7-5p 的靶标，GAB1 的上调消除了 miR-7-5p 过表达和 circ_0057452 敲低在 KFs 中的作用。Circ_0057452 通过吸附 miR-7-5p 在 KFs 中调节 GAB1 的表达。Circ_0057452 通过调节 miR-7-5p/GAB1 轴抑制瘢痕疙瘩的发展，这可能为瘢痕疙瘩提供一个有前途的治疗靶点。
