%0 Journal Article
%T 微小RNA222靶向调控基质金属蛋白酶1促进增生性瘢痕组织成纤维细胞生长
%A 张谊()
%A 张璃
%A 张启瑜
%A 洪炜龙
%A 林孝华
%A ()
%J 浙江大学学报(医学版)
%D 2017
%R 10.3785/j.issn.1008-9292.2017.12.06
%X 目的: 观察微小RNA（miR-）222在增生性瘢痕（HS）组织中的表达，并研究其调控成纤维细胞生长的机制。方法: 采用实时定量RT-PCR检测36例患者HS和正常组织中miR-222的表达；MTT法、流式细胞术和蛋白质印迹法分别检测成纤维细胞的增殖能力、生长周期、凋亡和相关蛋白表达。双荧光素酶报告分析确定miR-222的靶基因。结果: 与正常皮肤组织相比，HS组织中miR-222表达显著上调（P < 0.05）。上调miR-222的表达可显著提高成纤维细胞的增殖能力，增加增殖细胞核抗原（PCNA）、Ⅰ型和Ⅲ型胶原α1的mRNA和蛋白表达，上调S期细胞比例和细胞周期相关蛋白的表达，下调成纤维细胞的凋亡率和凋亡相关蛋白的表达。MMP1是miR-222的靶基因，miR-222通过绑定MMP1-3'UTR区负向调控MMP1在成纤维细胞的表达。MMP1可逆转miR-222的部分促纤维化作用。结论: miR-222通过负调控其靶基因MMP1的表达来实现其调控成纤维细胞的生长和凋亡。miR-222/MMP1信号通路可能成为HS诊断和治疗的生物学标志物和靶点。</br>Abstract: Objective: To explore the effect of microRNA(miR)-222 on cell proliferation and apoptosis of fibroblasts in hypertrophic scar (HS) and the underlying mechanisms. Methods: The expression of miR-222 in the HS and the normal skin tissues was detected by real-time RT-PCR. The HS fibroblasts were transfected with miR-222 mimic and miR-222 inhibitor respectively. The cell viability was tested with MTT assay, cell cycle distribution and apoptosis were detected with flow cytometry and the expression levels of proliferation, apoptosis and cell cycle related proteins were determined with Western blot. Direct target of miR-222 was evaluated by dual-luciferase reporter assay. Results: miR-222 was significantly up-regulated in HS tissues compared with normal skin tissues(P < 0.05). Overexpression of miR-222 enhanced the cell viability of HS fibroblasts; increased mRNA and protein expressions of proliferating cell nuclear antigen (PCNA), collagen alpha-1 (Ⅰ) chain (Col1A1) and collagen alpha-1 (Ⅲ) chain (Col3A1); increased cell population in S phase and protein expressions of cyclin D1, cyclin E1 and cyclin-dependent kinases 1 (CDK1); inhibited cell apoptosis and reduced protein expressions of caspase-3/9. Overexpression of MMP1 attenuated the effects of miR-222 on the cell viability and apoptosis in fibroblasts, reduced expression levels of PCNA, cyclin D1 and the expression of caspase-3 was increased. Conclusion: miR-222 enhances cell proliferation and inhibits cell apoptosis of HS fibroblasts through negative regulation of MMP1, which suggests that miR-222 and MMP1 might be used as novel biomarkers and targets in diagnostic and therapeutic approaches for HS. Key words: MicroRNAs/pharmacology Matrix metalloproteinase 1 Fibroblasts Cicatrix, hypertrophic/etiology Cicatrix, hypertrophic/therapy Cell proliferation
%K MicroRNAs/pharmacology Matrix metalloproteinase 1 Fibroblasts Cicatrix
%K hypertrophic/etiology Cicatrix
%K hypertrophic/therapy Cell proliferation
%U http://www.zjujournals.com/med/CN/10.3785/j.issn.1008-9292.2017.12.06