MicroRNA-491-5p is involved in the pathogenesis of degenerative lumbar scoliosis by targeting matrix metalloproteinase 9

doi:10.3969/j.issn.2095-4344.2016.02.017

Abstract

Abstract:

BACKGROUND: MicroRNAs are widely involved in the regulation of protein expression, and play a critical role in many physiological and pathological processes in the body. But microRNA expression profile in degenerative lumbar scoliosis is rarely reported and understood.
OBJECTIVE: To compare the microRNA expression profile in the normal intervertebral disc and degenerative lumbar scoliosis and to identify degenerative lumbar scoliosis-specific microRNAs, followed by functional validation.
METHODS: Total RNA samples were extracted from the nucleus pulposus tissues of 57 patients with degenerative lumbar scoliosis as experimental groups and the normal nucleus pulposus tissues of 42 patients with lumbar fractures as control group. An initial screening of differentially expressed microRNAs in the nucleus pulposus tissues by microRNA microarray was performed in 10 samples from each group. Subsequently, differentially expressed microRNAs were validated using real-time quantitative RCR. The level of differentially expressed microRNAs in the degenerative nucleus pulposus tissues was investigated. Then, the functional analysis of microRNAs in regulating collagen II expression was carried out. Western blot and luciferase reporter assay were also used to detect target genes.
RESULTS AND CONCLUSION: We identified 22 microRNAs that were differentially expressed (17 upregulated and 5 downregulated) in degenerative lumbar scoliosis patients compared with the controls. Following real-time quantitative RCR confirmation, miR-491-5p was significantly down-regulated in degenerative nucleus pulposus tissues in comparison with the controls. Moreover, its level was closely correlated with the pathological grading of disc degeneration. Overexpression of miR-491-5p promoted type II collagen expression in nucleus pulposus cells. Bioinformatics target prediction identified matrix metalloproteinase-9 as a putative target of miR-491-5p. Furthermore, luciferase reporter assays demonstrated that miR-491-5p directly targeted matrix metalloproteinase-9 and affected its protein expression in nucleus pulposus cells. These results show that the downregulation of miR-491-5p induces type II collagen loss by directly targeting matrix metalloproteinase-9, thereby resulting in degeneration of the intervertebral disc and degenerative lumbar scoliosis. This study also underscores the potential of miR-491-5p as a novel therapeutic target in degenerative lumbar scoliosis.

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

Wang Lei, Li Tian-wang, Liu Jian-qiang, Liu Xiao-zong, Wang Zhao-guo, Tian Yan, Zhang Yong-xing, Wang Wei . MicroRNA-491-5p is involved in the pathogenesis of degenerative lumbar scoliosis by targeting matrix metalloproteinase 9[J]. Chinese Journal of Tissue Engineering Research, 2016, 20(2): 248-253.

Figures/Tables 4

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