KCNQ1OT1 通过 miR-200b-3p/SERP1 轴介导角质形成细胞迁移，促进皮肤伤口愈合。

KCNQ1OT1 mediates keratinocyte migration to promote skin wound healing through the miR-200b-3p/SERP1 axis.

机构信息

Department of Plastic Surgery, Taizhou Central Hospital/Taizhou University Hospital, Taizhou, Zhejiang, China.

Department of Cardiothoracic Surgery, Taizhou Central Hospital/Taizhou University Hospital, Taizhou, Zhejiang, China.

出版信息

Burns. 2023 Mar;49(2):415-424. doi: 10.1016/j.burns.2022.04.011. Epub 2022 Apr 22.

DOI:10.1016/j.burns.2022.04.011

PMID:35523657

Abstract

BACKGROUND

The basic functions of keratinocyte are crucial steps during skin wound healing. KCNQ1OT1 long noncoding RNA was found to accelerate the migration and proliferation of keratinocyte in psoriasis. Here, we elucidated the action and mechanism of KCNQ1OT1 in skin wound healing.

METHODS

Expression levels of genes and proteins were evaluated by quantitative real-time PCR (qRT-PCR) and western blotting. Cell migration was assessed by using scratch and transwell assays. The interaction between miR-200b-3p and KCNQ1OT1 or SERP1 (Stress Associated Endoplasmic Reticulum Protein 1) was confirmed by bioinformatics analysis, dual-luciferase reporter assay, RNA immunoprecipitation (RIP) assay and pull-down assay.

RESULTS

KCNQ1OT1 had increased significantly in wound edge 1 day and 7 day after injury. Functionally, overexpression of KCNQ1OT1 promoted keratinocyte migration. Mechanistically, KCNQ1OT1/miR-200b-3p/SERP1 constituted a competing endogenous RNA (ceRNA) network in keratinocytes. A series of rescue experiments showed that miR-200b-3p up-regulation in keratinocytes attenuated the pro-migration action of KCNQ1OT1 in cells. Moreover, knockdown of miR-200b-3p could promote keratinocyte migration, which was abolished by SERP1 silencing. KCNQ1OT1 competitively sponged for miR-200b-3p to elevate the expression of its target SERP1.

CONCLUSION

KCNQ1OT1 could promote keratinocyte migration by miR-200b-3p/SERP1 axis, suggesting that KCNQ1OT1 might play a crucial role in skin wound healing.

摘要

背景

角质形成细胞的基本功能是皮肤伤口愈合过程中的关键步骤。现已发现 KCNQ1OT1 长非编码 RNA 可加速银屑病角质形成细胞的迁移和增殖。在这里，我们阐明了 KCNQ1OT1 在皮肤伤口愈合中的作用和机制。

方法

通过定量实时 PCR（qRT-PCR）和 Western blot 评估基因和蛋白质的表达水平。通过划痕和 Transwell 测定评估细胞迁移。通过生物信息学分析、双荧光素酶报告基因检测、RNA 免疫沉淀（RIP）实验和下拉实验证实 miR-200b-3p 与 KCNQ1OT1 或 SERP1（应激相关内质网蛋白 1）之间的相互作用。

结果

KCNQ1OT1 在受伤后 1 天和 7 天伤口边缘的表达显著增加。功能上，过表达 KCNQ1OT1 可促进角质形成细胞迁移。在机制上，KCNQ1OT1/miR-200b-3p/SERP1 在角质形成细胞中构成竞争性内源性 RNA（ceRNA）网络。一系列挽救实验表明，角质形成细胞中 miR-200b-3p 的上调可减弱 KCNQ1OT1 在细胞中的促迁移作用。此外，miR-200b-3p 的敲低可促进角质形成细胞迁移，而 SERP1 的沉默则可消除这一作用。KCNQ1OT1 通过竞争性吸附 miR-200b-3p 来提高其靶标 SERP1 的表达。