LINC01121 通过调节 miR-150-5p/MMP16 轴诱导椎间盘退变。
LINC01121 induced intervertebral disc degeneration via modulating miR-150-5p/MMP16 axis.
作者信息
Chen Xin, Li Zheng, Xu Derong, Li Shugang
机构信息
Department of Orthopaedic, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Graduate school of Peking Union Medical College, Beijing, 100042, China.
Department of Orthopedics, The Affiliated Hospital of Qingdao University, Qingdao, China.
出版信息
J Gene Med. 2020 Oct;22(10):e3231. doi: 10.1002/jgm.3231. Epub 2020 Jun 12.
BACKGROUND
Growing evidence indicates that Long noncoding RNAs contribute to cell differentiation, invasion, metabolism, proliferation and metastasis. However, the potential role of LINC01121 in progression of intervertebral disc degeneration (IDD) remains unclear.
METHODS
LINC01121, matrix metalloprotease (MMP)-16 and miR-150-5p expression was determined by a quantitative-reverse transcriptase-polymerase chain reaction assay. Inflammatory cytokines level was measured by an enzyme-linked immunosorbent assay and cell counting kit-8 analysis was used to assess cell proliferation. MMP-16-specific binding with miR-150-5p was verified with a luciferase reporter assay.
RESULTS
We noted that interleukin (IL)-1β and tumor necrosis factor (TNF)-α treatment enhanced LINC01121 and MMP-16 expression in nucleus pulposus (NP) cells. LINC01121 was higher in IDD specimens compared to that in control specimens. Higher expression of LINC01121 was correlated with disc degeneration degree. Ectopic expression of LINC01121 enhanced cell proliferation and promoted ki-67, MMP-3 and ADAMTS5 expression and also suppressed collagen II expression in NP cells. We observed that overexpression of LINC01121 increased the secretion of three inflammatory cytokines, including IL-6, TNF-α and IL-1β. We found that ectopic expression of LINC01121 decreased the miR-150-5p level in NP cells. Luciferase reporter data confirmed that MMP-16 was one direct target of miR-150-5p. Overexpression of miR-150-5p inhibited MMP-16 level and elevated the expression of LINC01121 enhanced MMP-16 level. We also found that MMP-16 was up-regulated in IDD specimens compared to that in control specimens. Higher expression of MMP-16 was correlated with disc degeneration degree. Interestingly, MMP-16 expression was positively related to LINC01121 in IDD specimens. Finally, overexpression of LINC01121 regulated cell growth, extracellular matrix degradation and inflammatory cytokine secretion via modulating MMP-16.
CONCLUSIONS
our data suggested LINC01121 may be a new therapeutic target for IDD.
背景
越来越多的证据表明，长链非编码RNA参与细胞分化、侵袭、代谢、增殖和转移。然而，LINC01121在椎间盘退变（IDD）进展中的潜在作用仍不清楚。
方法
采用定量逆转录聚合酶链反应法检测LINC01121、基质金属蛋白酶（MMP）-16和miR-150-5p的表达。采用酶联免疫吸附测定法检测炎性细胞因子水平，并用细胞计数试剂盒-8分析评估细胞增殖。通过荧光素酶报告基因测定法验证MMP-16与miR-150-5p的特异性结合。
结果
我们注意到白细胞介素（IL）-1β和肿瘤坏死因子（TNF）-α处理可增强髓核（NP）细胞中LINC01121和MMP-16的表达。与对照标本相比，IDD标本中LINC01121更高。LINC01121的高表达与椎间盘退变程度相关。LINC01121的异位表达增强了细胞增殖，促进了NP细胞中ki-67、MMP-3和ADAMTS5的表达，并抑制了胶原蛋白II的表达。我们观察到LINC01121的过表达增加了三种炎性细胞因子的分泌，包括IL-6、TNF-α和IL-1β。我们发现LINC01121的异位表达降低了NP细胞中miR-150-5p的水平。荧光素酶报告基因数据证实MMP-16是miR-150-5p的直接靶标之一。miR-150-5p的过表达抑制了MMP-16水平，而LINC01121的过表达增强了MMP-16水平。我们还发现，与对照标本相比，IDD标本中MMP-16上调。MMP-16的高表达与椎间盘退变程度相关。有趣的是，在IDD标本中，MMP-16表达与LINC01121呈正相关。最后，LINC01121的过表达通过调节MMP-16来调控细胞生长、细胞外基质降解和炎性细胞因子分泌。
结论
我们的数据表明LINC01121可能是IDD的一个新的治疗靶点。
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