Department of Plastic and Burn Surgery, The First Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu Province, China.
Bioengineered. 2022 Mar;13(3):7303-7315. doi: 10.1080/21655979.2022.2048771.
Scars are nearly impossible to avoid after a skin injury, but despite advancements in the treatment modalities, they remain a clinical problem, especially hypertrophic scars (HS). Many studies include the mechanism of formation and inhibition of HS, but it is not fully understood yet. Circular RNA HECTD1 (circHECTD1), for the first time, has been found to have roles in HS physiology. We determined the relative circHECTD1 levels in HS fibrous cells and tissues by RT-qPCR. Afterward, the effect of circHECTD1 knockdown on the proliferation, migration, invasion, fibrosis, and Transforming Growth Factor-beta/small mothers against decapentaplegic (TGF-β/Smad) signaling was studied using CCK-8, wound healing, Transwell, and western blot assays. After the role of circHECTD1 was clarified, its targeted micro RNA (miR) was predicted using the Starbase database, and we constructed a miR-142-3p mimic to study the details of its regulation mechanism. We used the TargetScan database to predict the downstream target high mobility group box 1 (HMGB1) of miR-142-3p, and the luciferase report assay verified the binding, and then its effect was determined by RT-qPCR. circHECTD1 is highly expressed in HS tissues and human skin hypertrophic scar fibroblasts (HSF); its loss of function inhibits cell proliferation, migration, invasion, fibrosis, and TGF-β/Smad signaling. However, miR-142-3p inhibitor reverses the effect of circHECTD1 on all the above-mentioned aspects, including HMGB1 expression. In conclusion, circHECTD1 knockdown interrupts TGF-β/Smad signaling through miR-142-3p/HMGB1 and suppresses scar fibrosis.
瘢痕在皮肤损伤后几乎无法避免，但尽管治疗方法有所进步，它们仍然是一个临床问题，尤其是增生性瘢痕（HS）。许多研究包括 HS 的形成和抑制机制，但尚未完全了解。环状 RNA HECTD1（circHECTD1）首次被发现具有 HS 生理学作用。我们通过 RT-qPCR 确定了 HS 纤维细胞和组织中的相对 circHECTD1 水平。之后，使用 CCK-8、划痕愈合、Transwell 和 Western blot 测定研究了 circHECTD1 敲低对增殖、迁移、侵袭、纤维化和转化生长因子-β/小 mothers against decapentaplegic（TGF-β/Smad）信号的影响。阐明 circHECTD1 的作用后，使用 Starbase 数据库预测其靶向 micro RNA（miR），并构建 miR-142-3p 模拟物以研究其调节机制的细节。我们使用 TargetScan 数据库预测 miR-142-3p 的下游靶 high mobility group box 1（HMGB1），并通过荧光素酶报告测定验证结合，然后通过 RT-qPCR 确定其作用。circHECTD1 在 HS 组织和人皮肤增生性瘢痕成纤维细胞（HSF）中高表达；其功能丧失抑制细胞增殖、迁移、侵袭、纤维化和 TGF-β/Smad 信号。然而，miR-142-3p 抑制剂逆转了 circHECTD1 对所有上述方面的作用，包括 HMGB1 表达。总之，circHECTD1 敲低通过 miR-142-3p/HMGB1 中断 TGF-β/Smad 信号通路，并抑制瘢痕纤维化。
