The First School of Clinical Medicine, Southern Medical University, Guangzhou 510010, China.
Department of Burn & Plastic Surgery, Guangzhou School of Clinical Medicine, Southern Medical University (General Hospital of Southern Theatre Command of PLA), Guangzhou 510010, China.
Burns. 2023 Aug;49(5):1157-1169. doi: 10.1016/j.burns.2022.07.010. Epub 2022 Jul 22.
Long non-coding RNA (lncRNA) dysregulation is demonstrated to be associated with disease progression. Mounting studies show that lncRNA promotes or inhibits the development of keloid. We aimed to disclose the role of homebox A11 antisense RNA (HOXA11-AS) in the formation of keloid.
Quantitative real-time PCR (qPCR) was adopted for expression analysis of HOXA11-AS, miR-182-5p and zinc finger protein 217 (ZNF217) mRNA, and the expression of ZNF protein and marker proteins was detected by western blot. Cell proliferation, cell migration and cell apoptosis were investigated using CCK-8 assay, wound healing assay and flow cytometry assay, respectively. The potential interplay between miR-182-5p and HOXA11-AS or ZNF217 was verified by dual-luciferase reporter assay, RIP assay and pull-down assay. The role of HOXA11 in vivo was studied by establishing animal models.
HOXA11-AS was highly expressed in tissues and fibroblasts of keloid. Deficiency of HOXA11-AS blocked the proliferation and migration of keloid fibroblasts and induced fibroblast apoptosis. HOXA11-AS directly combined to miR-182-5p whose downregulation reversed the effects of HOXA11-AS knockdown. ZNF217 was a target of miR-182-5p, and HOXA11-AS indirectly promoted ZNF217 expression by binding to miR-182-5p. MiR-182-5p enrichment also blocked keloid fibroblast proliferation, survival and migration, while further ZNF217 overexpression abolished these effects. HOXA11-AS knockdown also hindered the growth of keloid in mouse models.
High expression of HOXA11-AS promoted the formation and growth of keloid through the upregulation of ZNF217 by targeting miR-182-5p, and the inhibition of HOXA11-AS might be a novel strategy to prevent keloid development.
长链非编码 RNA（lncRNA）失调与疾病进展有关。越来越多的研究表明，lncRNA 促进或抑制瘢痕疙瘩的发展。我们旨在揭示同源盒 A11 反义 RNA（HOXA11-AS）在瘢痕疙瘩形成中的作用。
采用实时定量 PCR（qPCR）检测 HOXA11-AS、miR-182-5p 和锌指蛋白 217（ZNF217）mRNA 的表达，采用 Western blot 检测 ZNF 蛋白和标记蛋白的表达。采用 CCK-8 assay、划痕愈合assay 和流式细胞术assay 分别检测细胞增殖、细胞迁移和细胞凋亡。通过双荧光素酶报告基因assay、RIP assay 和 pull-down assay 验证 miR-182-5p 与 HOXA11-AS 或 ZNF217 之间的潜在相互作用。通过建立动物模型研究 HOXA11 在体内的作用。
HOXA11-AS 在瘢痕疙瘩组织和成纤维细胞中高表达。HOXA11-AS 缺失抑制瘢痕疙瘩成纤维细胞增殖和迁移，并诱导成纤维细胞凋亡。HOXA11-AS 直接与 miR-182-5p 结合，miR-182-5p 的下调逆转了 HOXA11-AS 敲低的作用。ZNF217 是 miR-182-5p 的靶标，HOXA11-AS 通过与 miR-182-5p 结合间接促进 ZNF217 表达。miR-182-5p 的富集也抑制了瘢痕疙瘩成纤维细胞的增殖、存活和迁移，而进一步过表达 ZNF217 则消除了这些作用。HOXA11-AS 敲低也抑制了小鼠模型中瘢痕疙瘩的生长。
高表达的 HOXA11-AS 通过靶向 miR-182-5p 上调 ZNF217 促进瘢痕疙瘩的形成和生长，抑制 HOXA11-AS 可能是预防瘢痕疙瘩发展的新策略。
