Department of Traumatic Ankle Surgery, Tianjin Hospital, Tianjin 300211, P.R. China.
Oncol Rep. 2018 Jun;39(6):2695-2702. doi: 10.3892/or.2018.6338. Epub 2018 Mar 28.
The roles of matrix metalloproteinase (MMP)9 in the control of pressure ulcers (PU) after hip fracture as well as how the rs1056629 in MMP9 3'UTR compromises the interaction between MMP9 and miR‑491 were explored. Online miRNA database (http://www.bioguo.org) was utilized to explore gene polymorphism in MMP9 3'UTR that might break the interaction between MMP9 and miRNA. Luciferase assay was utilized to confirm the miRNA targeted MMP9. Real‑time PCR, western blot analysis and immunohistochemistry were carried out to understand the roles of MMP9 in PU as well as how rs1056629 in MMP9 3'UTR compromises the interaction between MMP9 and miR‑491. rs1056629 in MMP9 3'UTR that compromised the interaction between MMP9 and four miRNAs including miR‑194‑3p, miR‑491, miR‑1915‑3p and miR‑941, and only miR‑491 among miR‑194‑3p, miR‑491, miR‑1915‑3p and miR‑941 decreased luciferase activity of wild‑type MMP9 3'UTR, and luciferase activities of mutant‑3 and mutant‑4 MMP9 3'UTR in miR‑491 overexpressing cells was comparable with scramble control. miR‑194‑3p, miR‑491, miR‑1915‑3p and miR‑941 levels in PU group was comparable with healthy control, and miR‑194‑3p, miR‑491, miR‑1915‑3p and miR‑941 in subjects carrying AA genotype was similar with those in AC and CC groups. MMP9 mRNA and protein, and histology score in subjects with PU were much higher, and were also much higher in AA group. Only miR‑491 mimic among miR‑194‑3p, miR‑491, miR‑1915‑3p and miR‑941 mimics downregulated the MMP9 level, and only miR‑491 inhibitor among miR‑194‑3p, miR‑491, miR‑1915‑3p and miR‑941 inhibitors upregulated the MMP9 level. Our study indicated that rs1056629 polymorphism could be a novel biomarker for predicting the occurrence of PU after a hip fracture.
探讨了基质金属蛋白酶（MMP）9 在髋部骨折后压疮（PU）中的作用，以及 MMP9 3'UTR 中的 rs1056629 如何影响 MMP9 与 miR-491 之间的相互作用。利用在线 miRNA 数据库（http://www.bioguo.org）探索 MMP9 3'UTR 中的基因多态性，这些多态性可能会破坏 MMP9 与 miRNA 之间的相互作用。利用荧光素酶报告基因检测证实 MMP9 是 miRNA 的靶基因。通过实时 PCR、western blot 分析和免疫组化检测 MMP9 在 PU 中的作用，以及 MMP9 3'UTR 中的 rs1056629 如何影响 MMP9 与 miR-491 之间的相互作用。MMP9 3'UTR 中的 rs1056629 影响 MMP9 与包括 miR-194-3p、miR-491、miR-1915-3p 和 miR-941 在内的四种 miRNA 之间的相互作用，其中只有 miR-491 降低了野生型 MMP9 3'UTR 的荧光素酶活性，并且在 miR-491 过表达的细胞中，突变型-3 和突变型-4 MMP9 3'UTR 的荧光素酶活性与 scramble 对照相似。PU 组与健康对照组相比，miR-194-3p、miR-491、miR-1915-3p 和 miR-941 的水平相似，携带 AA 基因型的受试者中 miR-194-3p、miR-491、miR-1915-3p 和 miR-941 的水平与 AC 和 CC 组相似。PU 患者的 MMP9 mRNA 和蛋白水平以及组织学评分均较高，AA 组也较高。在 miR-194-3p、miR-491、miR-1915-3p 和 miR-941 模拟物中，只有 miR-491 模拟物降低了 MMP9 水平，而在 miR-194-3p、miR-491、miR-1915-3p 和 miR-941 抑制剂中，只有 miR-491 抑制剂上调了 MMP9 水平。我们的研究表明，rs1056629 多态性可能是预测髋部骨折后发生 PU 的一种新的生物标志物。
