Zhang Qian, Guo Bingyu, Hui Qiang, Chang Peng, Tao Kai
Cell Physiol Biochem. 2018;49(3):985-995. doi: 10.1159/000493236. Epub 2018 Sep 5.
BACKGROUND/AIMS: There are few effective treatment options for hypertrophic scars. MircoRNAs are a class of small, noncoding RNAs involved in multiple biological functions.
Gene chip screening was used to screen out the differential expression of miRNAs in hypertrophic scars and normal tissues. Western blot and real-time PCR were used to confirm the expression of pleiotrophin (PTN) in hypertrophic scars. After analyze the correlation between PTN and miR-137 using correlation analysis, we used miRDB software to analyze the binding sites of miR-137 and PTN. Luciferase reporter gene, western blot and real-time PCR experiments were used to detect the regulatory effect of miR-137 on PTN. MTT and transwell assay were used to detect the effect of miR-137 on proliferation and metastasis. Western blot and real-time PCR were used to detect the regulatory effects of miR-137 on cyclin B1, matrix metalloproteinase 9 (MMP9), α-smooth muscle actin (α-SMA), vimentin, and type-I collagen (COL1A). Finally, miR-137 inhibitor was transfected into fibroblasts which was silent PTN, and the proliferation and migration of cells were detected by MTT and transwell. Western blot and real-time PCR were used to detect the expression of related proteins.
Various miRNAs was abnormal expressed in hypertrophic scars. miR-137 was decreased in hypertrophic scar, however PTN was up regulated in hypertrophic scars. miR-137 induced proliferation and metastasis in fibroblasts. This effect was accompanied by decreased expression of cyclin B1, MMP9, α-SMA, vimentin, and COL1A mediated via the target of PTN.
Modulation of miR-137 expression in fibroblasts could provide an important therapeutic strategy for hypertrophic scars.
背景/目的：肥厚性瘢痕的有效治疗方案较少。微小RNA是一类参与多种生物学功能的小的非编码RNA。
采用基因芯片筛选技术筛选肥厚性瘢痕组织与正常组织中差异表达的微小RNA。采用蛋白质免疫印迹法和实时荧光定量PCR法验证多效生长因子（PTN）在肥厚性瘢痕中的表达情况。通过相关性分析PTN与miR-137之间的相关性后，利用miRDB软件分析miR-137与PTN的结合位点。采用荧光素酶报告基因实验、蛋白质免疫印迹法和实时荧光定量PCR实验检测miR-137对PTN的调控作用。采用MTT法和Transwell实验检测miR-137对细胞增殖和转移的影响。采用蛋白质免疫印迹法和实时荧光定量PCR法检测miR-137对细胞周期蛋白B1、基质金属蛋白酶9（MMP9）、α-平滑肌肌动蛋白（α-SMA）、波形蛋白和I型胶原蛋白（COL1A）的调控作用。最后，将miR-137抑制剂转染至沉默PTN的成纤维细胞中，采用MTT法和Transwell实验检测细胞的增殖和迁移能力。采用蛋白质免疫印迹法和实时荧光定量PCR法检测相关蛋白的表达。
多种微小RNA在肥厚性瘢痕中表达异常。miR-137在肥厚性瘢痕中表达降低，而PTN在肥厚性瘢痕中表达上调。miR-137可诱导成纤维细胞增殖和转移。这种作用伴随着细胞周期蛋白B1、MMP9、α-SMA、波形蛋白和COLIA的表达降低，其是通过PTN靶点介导的。
调节成纤维细胞中miR-137的表达可为肥厚性瘢痕提供重要的治疗策略。
