miR-194-3p对瘢痕疙瘩成纤维细胞迁移的作用  

Effect of miR-194-3p on migration of keloid fibroblasts

作　　者：徐志山 回蔷[1] 李伟[1] 马书丹 郭冰玉[1] 常鹏[1] 陶凯[1] Xu Zhishan;Hui Qiang;Li Wei;Ma Shudan;Guo Bingyu;Chang Peng;Tao Kai(Department of Plastic Surgery,the General Hospital of Shenyang Military Region,Shenyang 110016,China)

机构地区：[1]沈阳军区总医院整形外科,110016

出　　处：《中华整形外科杂志》2018年第11期964-970,共7页Chinese Journal of Plastic Surgery

摘　　要：目的研究miR-194-3p对瘢痕疙瘩成纤维细胞迁移的影响。方法通过基因芯片方法,筛查出8份人瘢痕疙瘩与正常组织中差异性表达的miRNA。选取下调表达明显的miR-194-3p进行研究,通过miRDB预测其与RUNX2结合作用,并在人瘢痕疙瘩成纤维细胞(humankeloidfiroblasts,HKFs)及第3代瘢痕疙瘩细胞中,分别通过荧光报告基因验证miR-194-3p与RUNX2间的作用。在2种细胞中分别通过Transwell实验检测miR-194-3p对成纤维细胞迁移的作用。westernblot及real-timePCR分析HKFs细胞中miR-194-3p对RUNX2及迁移相关蛋白金属基质蛋白酶2(matrixmetallopeptidase2,MMP2)的作用。采用SPSS19.O软件对结果进行统计学分析,使用非配对t检验对各组进行比较,P<O.05为差异具有统计学意义。结果miR-721、miR-21-3p、miR-382-3p、miR-194-3p、miR-3107-5p和miR-144-5p在瘢痕疙瘩中存在异常表达的现象;miRDB软件分析显示,miR-194-3p与RUNX2存在靶向结合位点;报告基因实验证明,与对照组相比miR-194-3p能够抑制约50%的RUNX2活性(t=2.7764,P=O.0005)。基因和蛋白水平实验发现,miR-194-3p能够显著抑制RUNX2及MMP2的表达,进而抑制瘢痕疙瘩成纤维细胞的迁移(t=2.7764,P<0.05)。结论瘢痕疙瘩中miR-194-3p可能通过抑制RUNX2的活性来抑制成纤维细胞的迁移。Objective To study the effect of miR-194-3p on the migration of keloid fibroblasts.Methods Differentially expressed miRNA were screened by gene chip in 8 human keloid and normal tissues. The down regulated miR-194-3p was selected for study and its binding to RUNX2 was predicted by MiRDB, and verified by fluorescent reporter gene in human keloid fibroblasts (HKFs) and passage 3 keloid cells, respectively. The effect of miR-194-3p on the migration of fibroblasts was detected by transwell assay. Western blot and real-time PCR were used to analyze the effect of miR-194-3p on RUNX2 and MMP2 expression in HKFs. The results were analyzed by SPSS 19.0 software and compared by non-paired t test. P<0.05 was considered statistically significant.Results There were abnormal expressions of miR-721, miR-21-3p, miR-382-3p, miR-194-3p, miR-3107-5p and miR-144-5p in keloid. miRDB software analysis showed that miR-194-3p and RUNX2 had targeted binding sites. Reporter gene experiments showed that miR-194-3p inhibited the activity of RUNX2 by about 50% compared with the control group (t=2.7764, P=0.0005). MiR-194-3p could significantly inhibit the expression of RUNX2 and MMP2, and inhibited the migration of keloid fibroblasts (t=2.7764, P<0.05).Conclusion MiR-194-3p may inhibit the migration of fibroblasts by inhibiting the activity of RUNX2 in keloid.

关 键 词：瘢痕疙瘩 成纤维细胞 迁移 miR-194-3p RUNX2 

分 类 号：R622[医药卫生—整形外科]