| 摘要: |
| 目的观察成纤维胶原酶1（MMP1）和miR-222在增生性瘢痕（HS）成纤维细胞中的表达水平，探讨miR-222对HS发生、发展的调控机制。方法选取36例HS患者的HS组织，并各留取HS旁正常组织作为对照。分离培养出HS组织与正常组织的成纤维细胞；采用RT-PCR法检测成纤维细胞MMP1mRNA和miR-222表达水平；采用Westernblot法检测成纤维细胞MMP1蛋白表达水平；采用MTT法检测成纤维细胞增殖情况；采用双荧光素酶报告实验检测miR-222是否可调控MMP1基因。结果与正常组织比较，HS组织成纤维细胞MMP1mRNA、蛋白表达水平均下调（均P＜0.05），miR-222表达水平上调（P＜0.05）。与空白对照成纤维细胞比较，HS组织转染MMP1过表达质粒后的成纤维细胞MMP1mRNA、蛋白表达水平均上调（均P＜0.05），增殖速度明显减慢（P＜0.05）；转染antagomiR-222质粒后的成纤维细胞miR-222表达水平下调（P＜0.05），MMP1mRNA表达水平上调（P＜0.05），增殖速度明显减慢（P＜0.05）。双荧光素酶报告实验结果表明miR-222能与MMP1基因的3′-UTR区相结合，从而调控其表达。结论miR-222在HS组织中存在过表达，miR-222或通过负调控其靶基因MMP1的表达而发挥其调控成纤维细胞增殖和凋亡的作用。 |
| 关键词: 增生性瘢痕 microRNA-222 基质金属蛋白酶 1 |
| DOI：10.12056/j.issn.1006-2785.2017.39.24.2017-823 |
| 分类号: |
| 基金项目:浙江省医药卫生项目（2015KYB244） |
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| MicroRNA-222 regulates the proliferation of fibroblasts in hypertrophic scar via matrix metalloproteinase 1 |
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the First Affiliated Hospital of Wenzhou Medical University
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| Abstract: |
| Objective To determine the expressions of matrix metalloproteinase 1 (MMP1) and miR -222 in fibroblasts of hypertrophic scar (HS), and investigate the regulatory mechanism of miR-222 on the occurrence and development of HS. Methods HS tissues and HS-adjacent normal tissues were both collected from 36 HS patients. Primary fibroblasts were obtained. qRT-PCR was used to measure the level of mRNA of MMP1 and miR-222. Western blotting was carried out to determine MMP1 protein expression. MTT assay was employed to detect the proliferation of fibroblasts. Dual luciferase reporter assay was performed to identify the binding of miR-222 with MMP1 mRNA. Results MMP1 mRNA/protien were significantly downregulated
and miR-222 was significantly upregulated in HS fibroblasts, compared with control fibroblasts (P＜0.05). The expression of
MMP1mRNA/protein were significantly upregulated (P＜0.05),and the cell proliferation abilities was significantly inhibited in HS fibroblasts after transfected with pcDNAMMP1(P＜0.05). miR-222 was downregulated (P＜0.05), MMP1 mRNA was upregulated and the cell proliferation abilities was inhibited (P＜0.05)in HS fibroblasts after transfected with antagomiR-222,Dual-Luciferase Reporter assay showed miR-222 regulated the expression of MMP1 by binding with its 3′-UTR. Conclusion miR-222 overexpressed in HS fibroblasts, miR-222 may regulate the proliferation and apoptosis of fibroblasts in HS via negatively regulating the expression of target gene MMP1. |
| Key words: Hypertrophic scar MicroRNA-222 Matrix metalloproteinase 1 |
