Qi Jun, Liu Yifei, Hu Kesu, Zhang Yi, Wu Yangyang, Zhang Xia
Department of Burns and Plastic Surgery, Affiliated Hospital of Nantong University, Nantong, Jiangsu 226001, P.R. China.
Department of Pathology, Affiliated Hospital of Nantong University, Nantong, Jiangsu 226001, P.R. China.
Exp Ther Med. 2018 May;15(5):4332-4338. doi: 10.3892/etm.2018.5984. Epub 2018 Mar 21.
Hypertrophic scar (HS) is a fibrotic disease in which excessive extracellular matrix forms due to the response of fibroblasts to tissue damage. Novel evidence suggests that microRNAs (miRNAs or miRs) may contribute to hypertrophic scarring; however, the role of miRNAs in HS formation remains unclear. In the present study, miR-26a was significantly downregulated in HS tissues and human HS fibroblasts (hHSFs) was detected by reverse transcription-quantitative analysis. TargetScan was used to predict that mothers against decapentaplegic homolog 2 (Smad2) is a potential target gene of miR-26a and a dual-luciferase reporter assay confirmed that Smad2 was a target gene of miR-26a. The expression of Smad2 was upregulated in HS tissues and hHSFs. Cell Counting Kit-8 and flow cytometry analyses demonstrated that the overexpression of miR-26a significantly suppressed the proliferation ability of hHSFs and the apoptotic rate of hHSFs was significantly upregulated in response to miR-26a mimic transfection. Furthermore, the expression of B-cell lymphoma-2 (Bcl-2)-associated X protein was increased and Bcl-2 expression was decreased following miR-26a mimic transfection. The expression of collagens I and III was significantly inhibited following treatment with miR-26a mimics in hHSF cells. Conversely, miR-26a inhibitors served an opposing role in hHSFs. Furthermore, Smad2 overexpression enhanced the expression of collagens I and c III; however, Smad2 silencing inhibited the expression of collagens I and c III. In conclusion, the results of the present study indicate that miR-26a inhibits HS formation by modulating proliferation and apoptosis ad well as inhibiting the expression of extracellular matrix-associated proteins by targeting Smad2.
肥厚性瘢痕(HS)是一种纤维化疾病,其中由于成纤维细胞对组织损伤的反应而形成过多的细胞外基质。新证据表明,微小RNA(miRNA或miR)可能促成肥厚性瘢痕形成;然而,miRNA在HS形成中的作用仍不清楚。在本研究中,通过逆转录定量分析检测到miR-26a在HS组织和人HS成纤维细胞(hHSF)中显著下调。使用TargetScan预测,抗五肢瘫同系物2(Smad2)是miR-26a的潜在靶基因,双荧光素酶报告基因检测证实Smad2是miR-26a的靶基因。Smad2的表达在HS组织和hHSF中上调。细胞计数试剂盒-8和流式细胞术分析表明,miR-26a的过表达显著抑制hHSF的增殖能力,并且响应于miR-26a模拟物转染,hHSF的凋亡率显著上调。此外,miR-26a模拟物转染后,B细胞淋巴瘤-2(Bcl-2)相关X蛋白的表达增加,而Bcl-2表达降低。用miR-26a模拟物处理hHSF细胞后,I型和III型胶原蛋白的表达显著受到抑制。相反,miR-26a抑制剂在hHSF中起相反作用。此外,Smad过表达增强了I型和III型胶原蛋白的表达;然而,Smad2沉默抑制了I型和III型胶原蛋白的表达。总之,本研究结果表明,miR-26a通过调节增殖和凋亡以及通过靶向Smad2抑制细胞外基质相关蛋白的表达来抑制HS形成。