miR-155 promotes m6A modification of SOX2 mRNA through targeted regulation of HIF-1α and delays wound healing in diabetic foot ulcer in vitro models.

IF 3.2 3区 医学
Jiarui Peng, Hong Zhu, Bin Ruan, Zhisheng Duan, Mei Cao
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引用次数: 0

Abstract

Objective: Diabetic foot ulcers (DFU) are one of the most destructive complications of diabetes mellitus. The aim of this study was to link miR-155 and SOX2 with DFU to explore the regulation of wound healing by DFU and its potential mechanism.

Methods: Human keratinocytes (HaCaT) were induced with advanced glycation end products (AGEs) to construct DFU models in vitro. AGE-induced HaCaT cells were subjected to CCK-8 assays, flow cytometry, and wound healing assays to evaluate cell proliferation, apoptosis, and migration capacity, respectively. RT-qPCR and Western blotting were used to determine gene and protein expression levels, respectively. N6-methyladenosine (M6A) levels in total RNA were assessed using an M6A methylation quantification kit.

Results: Our results suggested that the inhibition of miR-155 promoted wound healing in an in vitro DFU model, while the knockdown of HIF-1α reversed this process, and that HIF-1α was a target protein of miR-155. In addition, knockdown of HIF-1α promoted the m6A level of SOX2 mRNA, inhibited the expression of SOX2, and inhibited the activation of the EGFR/MEK/ERK signaling pathway, thus inhibiting the proliferation and migration of HaCaT cells and promoting the apoptosis of HaCaT cells, while overexpression of SOX2 reversed this effect. We also found that METTL3 knockdown had the opposite effect of HIF-1α knockdown.

Conclusions: Inhibition of miR-155 promoted the expression of HIF-1α and attenuated the m6A modification of SOX2 mRNA, thereby promoting the expression of SOX2 and activating the downstream EGFR/MEK/ERK signaling pathway to promote wound healing in an in vitro DFU model.

miR-155 通过靶向调节 HIF-1α 促进 SOX2 mRNA 的 m6A 修饰,并延缓糖尿病足溃疡体外模型的伤口愈合。
目的:糖尿病足溃疡(DFU)是糖尿病最具破坏性的并发症之一:糖尿病足溃疡(DFU)是糖尿病最具破坏性的并发症之一。本研究旨在将 miR-155 和 SOX2 与 DFU 联系起来,探讨 DFU 对伤口愈合的调控及其潜在机制。方法:用高级糖化终产物(AGEs)诱导人角质形成细胞(HaCaT),在体外构建 DFU 模型。对 AGE 诱导的 HaCaT 细胞进行 CCK-8 试验、流式细胞术和伤口愈合试验,分别评估细胞增殖、凋亡和迁移能力。RT-qPCR 和 Western 印迹技术分别用于测定基因和蛋白质的表达水平。使用 M6A 甲基化定量试剂盒评估了总 RNA 中的 N6-甲基腺苷(M6A)水平:结果:我们的研究结果表明,在体外 DFU 模型中,抑制 miR-155 能促进伤口愈合,而敲除 HIF-1α 则能逆转这一过程,HIF-1α 是 miR-155 的靶蛋白。此外,HIF-1α的敲除促进了SOX2 mRNA的m6A水平,抑制了SOX2的表达,抑制了表皮生长因子受体/MEK/ERK信号通路的激活,从而抑制了HaCaT细胞的增殖和迁移,促进了HaCaT细胞的凋亡,而过表达SOX2则逆转了这一效应。我们还发现,METTL3敲除与HIF-1α敲除的效果相反:结论:在体外DFU模型中,抑制miR-155可促进HIF-1α的表达,减轻SOX2 mRNA的m6A修饰,从而促进SOX2的表达并激活下游的表皮生长因子受体/MEK/ERK信号通路,促进伤口愈合。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Journal of Diabetes Investigation
Journal of Diabetes Investigation Medicine-Internal Medicine
自引率
9.40%
发文量
218
期刊介绍: Journal of Diabetes Investigation is your core diabetes journal from Asia; the official journal of the Asian Association for the Study of Diabetes (AASD). The journal publishes original research, country reports, commentaries, reviews, mini-reviews, case reports, letters, as well as editorials and news. Embracing clinical and experimental research in diabetes and related areas, the Journal of Diabetes Investigation includes aspects of prevention, treatment, as well as molecular aspects and pathophysiology. Translational research focused on the exchange of ideas between clinicians and researchers is also welcome. Journal of Diabetes Investigation is indexed by Science Citation Index Expanded (SCIE).
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