Zhao Feng, Lang Hongxin, Wang Zhe, Zhang Tao, Zhang Dianbao, Wang Rui, Lin Xuewen, Liu Xiaoyu, Shi Ping, Pang Xining
Department of Stem Cells and Regenerative Medicine, Shenyang Key Laboratory for Stem Cells and Regenerative Medicine, Key Laboratory of Cell Biology, Ministry of Public Health, and Key Laboratory of Medical Cell Biology, Ministry of Education, China Medical University, 77 Puhe Street, Shenbei New District, Shenyang City 110122, Liaoning Province, China.
Department of Pathology, Shengjing Hospital of China Medical University, 36 Sanhao Street, Heping District, Shenyang City 110004, Liaoning Province, China.
Mol Ther Nucleic Acids. 2019 Mar 1;14:410-423. doi: 10.1016/j.omtn.2018.12.016. Epub 2019 Jan 10.
Early in gestation, wounds in fetal skin heal by regeneration, in which microRNAs play key roles. Seq-915_x4024 is a novel microRNA candidate confirmed by deep sequencing and mirTools 2.0. It is highly expressed in fetal keratinocytes during early gestation. Using an in vitro wound-healing assay, Transwell cell migration assay, and MTS proliferation assay, we demonstrated that keratinocytes overexpressing seq-915_x4024 exhibited higher proliferative activity and the ability to promote fibroblast migration and fibroblast proliferation. These characteristics of keratinocytes are the same biological behaviors as those of fetal keratinocytes, which contribute to skin regeneration. In addition, seq-915_x4024 suppressed the expression of the pro-inflammatory markers TNF-α, IL-6, and IL-8 and the pro-inflammatory chemokines CXCL1 and CXCL5. We also demonstrated that seq-915_x4024 regulates TGF-β isoforms and the extracellular matrix. Moreover, using an in vivo wound-healing model, we demonstrated that overexpression of seq-915_x4024 in keratinocytes suppresses inflammatory cell infiltration and scar formation. Using bioinformatics analyses, luciferase reporter assays, and western blotting, we further demonstrated that Sar1A, Smad2, TNF-α, and IL-8 are direct targets of seq-915_x4024. Furthermore, the expression of phosphorylated Smad2 and Smad3 was reduced by seq-915_x4024. Seq-915_x4024 could be used as an anti-fibrotic factor for the treatment of wound healing.
在妊娠早期，胎儿皮肤伤口通过再生愈合，其中微小RNA发挥关键作用。Seq-915_x4024是一种经深度测序和mirTools 2.0确认的新型微小RNA候选物。它在妊娠早期的胎儿角质形成细胞中高度表达。通过体外伤口愈合试验、Transwell细胞迁移试验和MTS增殖试验，我们证明过表达seq-915_x4024的角质形成细胞表现出更高的增殖活性以及促进成纤维细胞迁移和成纤维细胞增殖的能力。角质形成细胞的这些特性与胎儿角质形成细胞的生物学行为相同，有助于皮肤再生。此外，seq-915_x4024抑制促炎标志物TNF-α、IL-6和IL-8以及促炎趋化因子CXCL1和CXCL5的表达。我们还证明seq-915_x4024调节TGF-β异构体和细胞外基质。此外，使用体内伤口愈合模型，我们证明角质形成细胞中seq-915_x4024的过表达抑制炎症细胞浸润和瘢痕形成。通过生物信息学分析、荧光素酶报告基因试验和蛋白质印迹法，我们进一步证明Sar1A、Smad2、TNF-α和IL-8是seq-915_x4024的直接靶点。此外，seq-915_x4024降低了磷酸化Smad2和Smad3的表达。Seq-915_x4024可作为一种抗纤维化因子用于伤口愈合的治疗。
