Department of Plastic Surgery, The Third Hospital of Wuhan, Wuhan, Hubei 430060, P.R. China.
Department of Stomatology, Renmin Hospital of Wuhan University, Wuhan, Hubei 430060, P.R. China.
Mol Med Rep. 2021 Jan;23(1). doi: 10.3892/mmr.2020.11695. Epub 2020 Nov 17.
Keloids are a skin fibrotic disease that cause a number of problems for reconstructive surgeons. MicroRNAs (miRs) are crucial for the development of keloids. The present study aimed to investigate the function of the miR‑194‑5p/nuclear receptor subfamily 2 group F member 2 (NR2F2) interactome in human keloid fibroblasts. Microarray analysis was performed to identify key genes that may participate in keloid progression. The expression levels of miR‑194‑5p and NR2F2 mRNA in normal human skin fibroblasts (HSFs) and human keloid fibroblasts (KEL‑FIBs) were measured via reverse transcription‑quantitative PCR. Furthermore, cell proliferation, apoptosis, migration and invasion were assessed in KEL‑FIB cells. Following NR2F2 knockdown and miR‑194‑5p inhibition, NR2F2 expression was measured via western blotting. The microarray analysis identified NR2F2 as a key gene related to keloids. The regulatory association between miR‑194‑5p and NR2F2 was identified using TargetScan Human (version 7.2) and verified by performing a dual‑luciferase reporter assay. miR‑194‑5p expression was decreased in KEL‑FIB cells compared with HSF cells, and miR‑194‑5p overexpression inhibited the aggressive phenotypes of KEL‑FIB cells compared with the negative control group. Meanwhile, NR2F2 expression was negatively correlated with miR‑194‑5p expression. NR2F2 knockdown and miR‑194‑5p overexpression displayed similar effects on KEL‑FIB cells. Moreover, NR2F2 knockdown effectively reversed miR‑194‑5p inhibitor‑mediated effects in keloid fibroblasts. The present study indicated that the novel miR/194‑5p/NR2F2 interactome might be involved in the progression of keloid aggression and may serve as a potential therapeutic target for human keloid in the future.
瘢痕疙瘩是一种皮肤纤维组织疾病,会给整形医生带来诸多问题。微小 RNA(miRNA)在瘢痕疙瘩的发生发展中起着至关重要的作用。本研究旨在探讨miR-194-5p/核受体亚家族 2 组 F 成员 2(NR2F2)相互作用体在人瘢痕疙瘩成纤维细胞中的功能。通过微阵列分析鉴定可能参与瘢痕疙瘩进展的关键基因。采用逆转录定量 PCR 检测正常皮肤成纤维细胞(HSFs)和人瘢痕疙瘩成纤维细胞(KEL-FIBs)中 miR-194-5p 和 NR2F2 mRNA 的表达水平。此外,还评估了 KEL-FIB 细胞的增殖、凋亡、迁移和侵袭。NR2F2 敲低和 miR-194-5p 抑制后,通过 Western blot 检测 NR2F2 的表达。微阵列分析确定 NR2F2 是与瘢痕疙瘩相关的关键基因。通过 TargetScan Human(版本 7.2)鉴定 miR-194-5p 与 NR2F2 的调控关系,并通过双荧光素酶报告基因检测进行验证。与 HSF 细胞相比,KEL-FIB 细胞中 miR-194-5p 的表达降低,与阴性对照组相比,miR-194-5p 过表达抑制了 KEL-FIB 细胞的侵袭表型。同时,NR2F2 的表达与 miR-194-5p 的表达呈负相关。NR2F2 敲低和 miR-194-5p 过表达对 KEL-FIB 细胞的作用相似。此外,NR2F2 敲低可有效逆转 miR-194-5p 抑制剂对瘢痕疙瘩成纤维细胞的作用。本研究表明,新型 miR/194-5p/NR2F2 相互作用体可能参与瘢痕疙瘩侵袭的进展,有望成为未来人类瘢痕疙瘩的潜在治疗靶点。