Department of Plastic Surgery, Lanzhou University Second Hospital, Lanzhou, PR China.
J Dermatolog Treat. 2021 Dec;32(8):973-982. doi: 10.1080/09546634.2020.1728210. Epub 2020 Feb 21.
This study explored the function and mechanism of miR-486-5p in HSFBs.
Qualitative real-time-polymerase chain reaction (qRT-PCR) was performed to detect the expression of miR-486-5p in HS and hypertrophic scar fibroblasts (HSFBs). Viability, migration, invasion ability, apoptosis, and expressions of Collagen I, Collagen III, α-SMA and Cleaved caspase-3 in HSFBs after transfection with miR-486-5p mimic or inhibitor were measured by CCK-8, wound-healing, transwell, and Western blot, respectively. Interaction between miR-486-5p and IGF1 was predicted by Targetscan version 7.2 and further confirmed by dual-luciferase assay, and functional rescue experiments were conducted to verify the predicted molecular mechanism. The activation of PI3K/AKT pathway was also analyzed by Western blot.
MiR-486-5p was low-expressed in HS and HSFBs, and that overexpression of miR-486-5p suppressed the viability, migration, invasion, and expressions of Collagen I, Collagen III, and α-SMA of HSFBs, meanwhile, it also promoted apoptosis and Cleaved caspase-3 expression in HSFBs. Moreover, IGF1 was targeted by miR-486-5p, and increased viability, migration, invasion, and collagens expressions, the activation of PI3K/Akt pathway, and decreased apoptosis and Cleaved caspase-3 induced by miR-486-5p inhibitor could be partly alleviated by siIGF1.
Overexpressed miR-486-5p inhibited the hyperproliferation and excessive production of collagen in HSFBs IGF1/PI3K/AKT pathway.
本研究旨在探讨 miR-486-5p 在 HSFBs 中的功能和作用机制。
采用实时荧光定量聚合酶链反应（qRT-PCR）检测 HS 和增生性瘢痕成纤维细胞（HSFBs）中 miR-486-5p 的表达。通过 CCK-8 法、划痕实验、Transwell 实验和 Western blot 法分别检测转染 miR-486-5p 模拟物或抑制剂后 HSFBs 的活力、迁移、侵袭能力、凋亡以及 Collagen I、Collagen III、α-SMA 和 Cleaved caspase-3 的表达。通过 Targetscan version 7.2 预测 miR-486-5p 与 IGF1 的相互作用，并通过双荧光素酶报告基因实验进一步验证，同时进行功能拯救实验以验证预测的分子机制。还通过 Western blot 分析 PI3K/AKT 通路的激活情况。
miR-486-5p 在 HS 和 HSFBs 中低表达，过表达 miR-486-5p 抑制 HSFBs 的活力、迁移、侵袭以及 Collagen I、Collagen III 和 α-SMA 的表达，同时促进 HSFBs 的凋亡和 Cleaved caspase-3 的表达。此外，IGF1 是 miR-486-5p 的靶标，增加 miR-486-5p 抑制剂诱导的活力、迁移、侵袭和胶原表达、PI3K/Akt 通路的激活以及凋亡和 Cleaved caspase-3 的减少可部分被 siIGF1 缓解。
过表达 miR-486-5p 通过 IGF1/PI3K/AKT 通路抑制 HSFBs 的过度增殖和胶原过度产生。
