Downregulated Hsa-let-7f contributes to the loss of type II collagen by targeting interleukin-10/STAT3 signaling pathway in degenerative lumbar scoliosis

doi:10.3969/j.issn.2095-4344.2016.15.014

Abstract

Abstract:

BACKGROUND: MicroRNAs (miRNAs) play an important role in a variety of diseases. Investigation of miRNA expression profile in degenerative lumbar scoliosis is beneficial for understanding its pathogenesis, providing a novel therapeutic target. Therefore, we tested the hypothesis that miRNAs promote intervertebral disc degeneration through the interleukin-10/STAT3 signaling pathway, a potential regulator of intervertebral disc degeneration.
OBJECTIVE: To compare the differentially expressed miRNAs in the intervertebral disc tissues from patients with degenerative lumbar scoliosis and normal controls and to identify specific miRNAs in degenerative lumbar scoliosis followed by functional validation.
METHODS: An initial screening of miRNA expression in nucleus pulposus tissues by miRNA Solexa Sequencing was performed in samples from 10 patients with degenerative lumbar scoliosis and 10 controls, respectively. Subsequently, differentially expressed miRNAs were validated using qRT-PCR. The level of differentially expressed miRNAs in degenerative nucleus pulposus tissues was investigated. Then, functional analysis of the miRNAs in regulating type II collagen expression was carried out. Western blot and luciferase reporter assay were used to further confirm the target gene.
RESULTS AND CONCLUSION: We identified 30 miRNAs that were differentially expressed (16 upregulated and 14 downregulated) in patients with degenerative lumbar scoliosis compared with controls. Following qRT-PCR confirmation, Has-let-7f was significantly down-regulated in degenerative nucleus pulposus tissues as compared with controls. Moreover, its level was correlated with the severity of disc degeneration. Overexpression of Has-let-7f promoted type II collagen expression in nucleus pulposus cells. Knockout of interleukin-10 induced effects on nucleus pulposus cells similar to Has-let-7f. Bioinformatics target prediction identified interleukin-10 as a putative target of Has-let-7f. Furthermore, luciferase reporter assays demonstrated that Has-let-7f altered the expression of STAT3 and matrix metalloproteinase-2. These findings indicate that the downregulation of Has-let-7f induces type II collagen loss by directly targeting inleukin-10, thereby resulting in intervertebral disc degeneration and degenerative lumbar scoliosis. Has-let-7f is likely to be a novel therapeutic target for 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

Key words: Scoliosis, MicroRNAs, Interleukin-10, Tissue Engineering

Wang Lei, Li Tian-wang, Liu Jian-qiang, Liu Xiao-zong, Wang Zhao-guo, Tian Yan, Zhang Yong-xing, Wang Wei. Downregulated Hsa-let-7f contributes to the loss of type II collagen by targeting interleukin-10/STAT3 signaling pathway in degenerative lumbar scoliosis[J]. Chinese Journal of Tissue Engineering Research, 2016, 20(15): 2225-2232.

Figures/Tables 4

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