| 张爱新,张卓,张杰.miR-363在脆性骨折患者血中的表达及对MC3T3-E1细胞成骨分化及增殖的影响.[J].中南医学科学杂志.,2022,(2):206-210. |
| miR-363在脆性骨折患者血中的表达及对MC3T3-E1细胞成骨分化及增殖的影响 |
| Expression of miR-363 in blood of patients with brittle fracture and its effect on osteogenic differentiation and proliferation of MC3T3-E1 cells |
| 投稿时间：2021-04-14 修订日期：2021-10-19 |
| DOI：10.15972/j.cnki.43-1509/r.2022.02.012 |
| 中文关键词: miR-363 小鼠胚胎成骨细胞前体细胞 成骨分化 Dickkopf相关蛋白1 Wnt/β-catenin |
| 英文关键词:miR-363 MC3T3-E1 osteogenic differentiation DKK1 Wnt/β-catenin |
| 基金项目: |
|
| 摘要点击次数: 382 |
| 全文下载次数: 201 |
| 中文摘要: |
| 目的研究miR-363在脆性骨折患者血中的表达及对小鼠胚胎成骨细胞前体细胞(MC3T3-E1)成骨分化与增殖的影响。方法收集脆性骨折患者外周血,采用荧光定量PCR检测患者miR-363、矮小相关转录因子2(RUNX2)、骨钙素(OCN)、细胞周期蛋白D1(CCND1)、连环蛋白β1(CTNNB1)和骨髓细胞瘤癌基因(MYC)表达。采用茜素红S染色与噻唑蓝检测过表达miR-363对细胞成骨分化及增殖的影响。采用双荧光素酶报告基因与Western blotting检测miR-363对Dickkopf相关蛋白1(DKK1)的调控作用。结果miR-363在脆性骨折患者外周血中的表达水平低于正常健康者,而术后1周、2周及4周外周血中的表达水平高于术前并呈上调趋势(P＜0.05)；模拟物组MC3T3-E1细胞中miR-363表达水平高于阴性对照组(P＜0.05)；模拟物组MC3T3-E1细胞矿化程度、RUNX2及OCN mRNA表达水平及1天、2天、3天光密度(OD)值高于阴性对照组(P＜0.05)；DKK1是miR-363的靶基因,模拟物组MC3T3-E1细胞中DKK1蛋白表达水平低于阴性对照组,而CCND1、CTNNB1及MYC mRNA表达水平高于阴性对照组(P＜0.05)。结论miR-363可能通过调控靶基因DKK1与Wnt/β-catenin信号通路促进MC3T3-E1细胞成骨分化与增殖。 |
| 英文摘要: |
| To investigate the expression of miR-363 in the blood of patients with brittle fracture and the effect of miR-363 on the osteogenic differentiation and proliferation of mouse embryonic osteoblast precursor cells (MC3T3-E1). MethodsThe peripheral blood of patients with fragility fractures were collected, and the expression of miR-363, runt-related transcription factor 2 (RUNX2), osteocalcin (OCN), cyclin D1 (CCND1), catenin beta 1 (CTNNB1) and myelocytomatosis oncogene (MYC) were detected by fluorescence quantitative PCR. The effect of overexpression of miR-363 on osteogenic differentiation and proliferation of cells was detected by alizarin red S staining and thiazole blue.The regulation of miR-363 on Dickkopf-1 (DKK1) was detected by dual luciferase reporter gene and Western blotting. ResultsThe expression level of miR-363 in peripheral blood of fragility fracture patients was lower than that of normal healthy individuals, but its expression level in peripheral blood of these patients at 1 week, 2 weeks, and 4 weeks after operation was higher than that of pre-operation and showed an upward-regulated trend (P＜0.05); the expression level of miR-363 in MC3T3-E1 cells in the mimic group was higher than that in the negative control group (P＜0.05); the degree of mineralization, the mRNA expression levels of RUNX2 and OCN, and optical density (OD) at 1 day, 2 days, and 3 days in MC3T3-E1 cells in the mimic group were higher than those in the negative control group (P＜0.05); DKK1 is the target gene of miR-363, the expression level of DKK1 protein in MC3T3-E1 cells in the mimic group was lower than that in the negative control group, while the mRNA expression levels of CCND1, CTNNB1 and MYC were higher than that in the negative control group (P＜0.05). ConclusionmiR-363 may promote osteogenic differentiation and proliferation of MC3T3-E1 cells by regulating target gene DKK1 and Wnt/β-catenin signaling pathway. |
| 查看全文 查看/发表评论 下载PDF阅读器 |
| 关闭 |
|
|
|