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微小RNA-497-5p表达下调通过调节Smad7抑制人增生性瘢痕成纤维细胞的细胞活力,减少细胞外基质沉积并诱导细胞凋亡。

MicroRNA-497-5p downregulation inhibits cell viability, reduces extracellular matrix deposition and induces apoptosis in human hyperplastic scar fibroblasts by regulating Smad7.

作者信息

Li Zhiqiang, Wang Pengtao, Zhang Jie, Zhao Dongkui

机构信息

Shanghai Meizhizhen Medical Cosmetology Clinic, Shanghai 200122, P.R. China.

出版信息

Exp Ther Med. 2021 Apr;21(4):384. doi: 10.3892/etm.2021.9815. Epub 2021 Feb 23.

Abstract

Hypertrophic scars (HSs) are characterized by excessive extracellular matrix deposition and excessive growth of dense fibrous tissues. MicroRNAs (miRNAs/miRs) serve key roles in HS formation. The present study investigated the expression, role and mechanism underlying the effects of miR-497-5p in HS formation. miR-497-5p expression was detected via reverse transcription-quantitative PCR. The association between miR-497-5p and Smad7 was analyzed using TargetScan and luciferase reporter assays. Protein expression levels of extracellular matrix markers were measured via western blotting. Cell viability and apoptosis were determined using the Cell Counting Kit-8 assay and flow cytometry, respectively. The results suggested that miR-497-5p expression was upregulated in HS tissues and human HS fibroblasts (hHSFs) compared with healthy control skin tissues and CCC-ESF-1 cells, respectively. Smad7 was directly targeted by miR-497-5p, and was downregulated in HS tissues and hHSFs compared with healthy control skin tissues and CCC-ESF-1 cells, respectively. Moreover, Smad7 upregulation significantly inhibited cell viability, decreased extracellular matrix deposition and induced apoptosis in hHSFs compared with the control-plasmid group. Moreover, the results indicated that, compared with the inhibitor control group, miR-497-5p inhibitor inhibited cell viability, decreased extracellular matrix deposition and induced apoptosis in hHSFs, which were significantly reversed by Smad7 knockdown. In conclusion, the results indicated that miR-497-5p downregulation repressed HS formation by inhibiting extracellular matrix deposition and hHSF proliferation at least partly by targeting Smad7.

摘要

肥厚性瘢痕(HSs)的特征是细胞外基质过度沉积和致密纤维组织过度生长。微小RNA(miRNAs/miRs)在HS形成中起关键作用。本研究调查了miR-497-5p在HS形成中的表达、作用及潜在机制。通过逆转录定量PCR检测miR-497-5p的表达。使用TargetScan和荧光素酶报告基因检测分析miR-497-5p与Smad7之间的关联。通过蛋白质印迹法测量细胞外基质标志物的蛋白质表达水平。分别使用细胞计数试剂盒-8检测法和流式细胞术测定细胞活力和细胞凋亡。结果表明,与健康对照皮肤组织和CCC-ESF-1细胞相比,HS组织和人HS成纤维细胞(hHSFs)中miR-497-5p的表达分别上调。Smad7是miR-497-5p的直接靶标,与健康对照皮肤组织和CCC-ESF-1细胞相比,HS组织和hHSFs中Smad7的表达分别下调。此外,与对照质粒组相比,Smad7上调显著抑制hHSFs的细胞活力,减少细胞外基质沉积并诱导细胞凋亡。此外,结果表明,与抑制剂对照组相比,miR-497-5p抑制剂抑制hHSFs的细胞活力,减少细胞外基质沉积并诱导细胞凋亡,而Smad7基因敲低可显著逆转这些作用。总之,结果表明miR-497-5p下调至少部分通过靶向Smad7抑制细胞外基质沉积和hHSF增殖,从而抑制HS形成。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/39a4/7918061/16b54e0e05ea/etm-21-04-09815-g00.jpg

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