miR-524通过Egr1抑制增生性瘢痕成纤维细胞的生长  

MiR-524 inhibits the growth of hypertrophic scar fibroblasts by inhibiting Egr1

作　　者：付志强 白泽明 陶凯 FU Zhiqiang;BAI Zeming;TAO Kai(Department of Burns and Plastic Surgery,General Hospital of Northern Theater Command,Shenyang 110016,China)

机构地区：[1]北部战区总医院整形外科,辽宁沈阳110016

出　　处：《中国美容整形外科杂志》2022年第4期232-235,I0004,共5页Chinese Journal of Aesthetic and Plastic Surgery

摘　　要：目的探讨miR-524是否能够通过调节Egr1影响增生性瘢痕成纤维细胞的生长。方法自2018年9月至2020年6月,北部战区总医院烧伤整形科通过定量逆转录聚合酶链反应检测正常组织和增生性瘢痕组织中miR-524和Egr1的表达情况。获取原代增生性瘢痕成纤维细胞,将其分为三组,分别转染无意义对照链(对照组)、miR-524(miR-524组)、miR-524抑制物(miR-524抑制物组),在0、1、2、3、4 d通过CCK-8实验检测细胞的增殖情况,转染后24 h,采用细胞周期试剂盒检测细胞周期,细胞凋亡试剂盒检测细胞凋亡。STARBASE软件预测miR-524与Egr1结合位点。转染后24 h蛋白免疫印记实验检测Egr1、Cyclin D1和p21表达。结果q RT-PCR实验结果表明,miR-524在增生性瘢痕组织中的相对表达含量为0.568±0.085,在正常组织中的相对表达含量为0.967±0.009,miR-524在增生性瘢痕组织中表达显著低于正常组织(t=7.577,P=0.017)。Egr1在增生性瘢痕组织中的表达含量(1.360±0.120)显著高于正常组织(0.982±0.012,t=5.013,P=0.038)。miR-524转染后能够显著抑制细胞的增殖能力,miR-524过表达能够抑制细胞周期(G1-S期)的转化,与对照组相比miR-524转染后细胞凋亡比例增加。通过生物学软件预测发现,miR-524与Egr1具有结合位点。与对照组相比,miR-524能够下调Egr1、Cyclin D1和p21表达,添加miR-524抑制物后Egr1、Cyclin D1和p21表达上调。结论miR-524通过抑制增生性瘢痕成纤维细胞中的Egr1而抑制增生性瘢痕成纤维细胞的增殖、促进其凋亡。Objective To explore the effect of miR-524 on the growth of hypertrophic scar fibroblasts by regulating Egr1.Methods From September 2018 to June 2020,the expression of miR-524 and Egr1 in normal tissues and hypertrophic scars was detected by quantitative reverse transcription polymerase chain reaction(q RT-PCR).Primary hypertrophic scar fibroblasts were obtained and divided into 3 groups.These fibroblasts were transfected with meaningless chain(control group),miR-524(miR-524 group)and miR-524 inhibitor(miR-524 inhibitor group).The proliferation of primary fibroblasts at 0,1,2,3 and 4 days was detected by CCK8 assay.The cell cycle was detected by cell cycle kit and apoptosis was detected by cell apoptosis kit at 24 h after transfection.The binding sites of miR-524 and Egr1 were predicted by Starbase software.The expression of Egr1,cyclin D1 and p21 at 24 h after transfection was detected by Western blot.Results The results of q RT-PCR showed that the relative expression level of miR-524 in hypertrophic scars was 0.568±085,which significantly lower than that in normal tissues of 0.967±0.009(t=7.577,P=0.017).The expression level of Egr1 in hypertrophic scars was1.360±0.120,which was significantly higher than that in normal tissues of 0.982±0.012(t=5.013,P=0.038).MiR-524 transfection could significantly inhibit fibroblasts proliferation and miR-524 overexpression could inhibit G1/S phase transition in the cell cycle.Compared with the control group,the proportion of apoptosis increased after miR-524 transfection.The binding sites of miR-524 and Egr1 were predicted by biological software.Compared with the control group,miR-524 could down regulate the expression of Egr1,cyclin D1 and p21 and up regulate the expression of Egr1,cyclin D1 and p21 after adding miR-524 inhibitor.Conclusion MiR-524 could inhibit proliferation and promote apoptosis of hypertrophic scar fibroblasts by inhibiting Egr1.

关 键 词：增生性瘢痕 成纤维细胞 微小RNA 早期生长反应因子-1 

分 类 号：R622[医药卫生—整形外科]