miR-520h对缺氧状态下人牙周膜细胞MMP-2表达的影响

发布时间：2018-05-07 08:15

本文选题：miR-h + 缺氧　；参考：《实用口腔医学杂志》2014年03期

【摘要】：目的:探讨miR-520h对缺氧状态下人牙周膜细胞(HPDLCs)MMP-2表达的影响。方法:将HPDLCs置于1%氧浓度的孵箱中培养0、12、24、48和72 h。用实时荧光定量RT-PCR(qRT-PCR)和Western blot检测细胞中MMP-2 mRNA和蛋白的表达。构建野生型和突变型MMP-2表达载体,用TargetScan和miRanda等软件预测作用于MMP-2基因3'-UTR靶向miRNA,双荧光素酶报告基因检测系统验证靶向MMP-2 3'-非翻译区(3'-UTR)的miRNA。用qRT-PCR和Western blot技术检测miR-520h对MMP-2表达的调控。结果:HPDLCs在缺氧培养中MMP-2 mRNA和蛋白的表达量明显下调(P0.01)。软件预测在MMP-2的3'-UTR 516bp位置有一个与miR-520h的结合位点,HPDLCs被转染pre-miR-520后,其表达量增高(P0.01),MMP-2载体的荧光素酶活性降低(P0.05)。miR-520h都抑制了HPDLCs中MMP-2 mRNA和蛋白表达(P0.05)。结论:miR-520h可抑制缺氧状态下牙周膜细胞MMP-2基因表达。
[Abstract]:Aim: to investigate the effect of miR-520h on the expression of MMP-2 in human periodontal ligament cells under hypoxia. Methods: HPDLCs was incubated in 1% oxygen incubator for 48 and 72 hours. The expression of MMP-2 mRNA and protein was detected by real-time quantitative RT-PCR and Western blot. The wild-type and mutant MMP-2 expression vectors were constructed, and TargetScan and miRanda were used to predict the effect of MMP-2 gene 3'-UTR targeting miRNAs. The double-luciferase reporter gene detection system was used to verify the MIRNAs targeting the MMP-2 _ (3) -untranslated region. The regulation of MMP-2 expression by miR-520h was detected by qRT-PCR and Western blot techniques. Results the expression of MMP-2 mRNA and protein in hypoxic culture was significantly down-regulated (P 0.01). The software predicted that at the 3'-UTR 516bp site of MMP-2, there was a binding site for miR-520h and HPDLCs were transfected into pre-miR-520, and the expression level of P0.01MMPP-2 was increased. The decrease of luciferase activity of P0.05N. MiR-520h inhibited the expression of MMP-2 mRNA and protein in HPDLCs. Conclusion the expression of MMP-2 gene in periodontal ligament cells can be inhibited by 10% miR-520h.
【作者单位】：第三军医大学新桥医院口腔科;
【基金】：国家自然科学基金(编号:81170934) 第三军医大学青年创新基金(编号:2011D274)
【分类号】：R781.4

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