Yuan Xiaoye, Chen Baiye, Wang Xueming
Department of Plastic Surgery, Fujian Maternal and Child Health Hospital, Fuzhou, Fujian 350000, China.
Department of Pediatric Burn and Plastic Surgery, Fujian Children's Hospital, Fuzhou, Fujian 350000, China.
Burns. 2023 May;49(3):622-632. doi: 10.1016/j.burns.2022.04.009. Epub 2022 Apr 22.
Circular RNAs (circRNAs) are identified as important regulators in human diseases, including keloid. The purpose of this study is to reveal the role and molecular mechanism of circSLC8A1 in keloid formation.
Expression of circSLC8A1, microRNA (miR)-181a-5p, and hypoxia inducible factor 1 alpha inhibitor (HIF1AN) were detected by quantitative real-time PCR. Protein expression of extracellular matrix (ECM) deposition markers and HIF1AN was detected by western blot analysis. Furthermore, the interaction between miR-181a-5p and circSLC8A1 or HIF1AN was confirmed by dual-luciferase reporter assay, RIP assay and RNA pull-down assay.
Expression of circSLC8A1 was downregulated in keloid tissues and HKFs. Overexpression of circSLC8A1 suppressed HKFs proliferation, migration, ECM deposition, and promoted apoptosis. MiR-181a-5p is targeted by circSLC8A1, and its mimic reversed the effect of circSLC8A1 on the biological function of HKFs. HIF1AN was a target of miR-181a-5p, and it was positively regulated by circSLC8A1. Knockdown of HIF1AN also reversed the negatively regulation of circSLC8A1 on the biological functions of HKFs.
Our data showed that circSLC8A1 regulates the miR-181a-5p/HIF1AN axis to restrain HKFs biological functions, confirming that circSLC8A1 might serve as a novel therapeutic target for keloids.
环状RNA（circRNAs）被认为是人类疾病（包括瘢痕疙瘩）中的重要调节因子。本研究的目的是揭示circSLC8A1在瘢痕疙瘩形成中的作用和分子机制。
通过定量实时PCR检测circSLC8A1、微小RNA（miR）-181a-5p和缺氧诱导因子1α抑制剂（HIF1AN）的表达。通过蛋白质免疫印迹分析检测细胞外基质（ECM）沉积标志物和HIF1AN的蛋白表达。此外，通过双荧光素酶报告基因检测、RNA免疫沉淀实验（RIP实验）和RNA下拉实验证实miR-181a-5p与circSLC8A1或HIF1AN之间的相互作用。
circSLC8A1在瘢痕疙瘩组织和人瘢痕疙瘩成纤维细胞（HKFs）中表达下调。circSLC8A1的过表达抑制了HKFs的增殖、迁移、ECM沉积，并促进了细胞凋亡。miR-181a-5p是circSLC8A1的靶标，其模拟物可逆转circSLC8A1对HKFs生物学功能的影响。HIF1AN是miR-181a-5p的靶标，且受circSLC8A1的正向调控。敲低HIF1AN也可逆转circSLC8A1对HKFs生物学功能的负向调控。
我们的数据表明，circSLC8A1通过调节miR-181a-5p/HIF1AN轴来抑制HKFs的生物学功能，证实circSLC8A1可能是瘢痕疙瘩的一个新的治疗靶点。
