Jing Wanli, Liu Wei
Department of Orthopaedics, Tianjin First Central Hospital, Tianjin, China.
Department of Orthopaedics, Baodi Peopele's Hospital, Tianjin, China.
Front Mol Biosci. 2021 Jul 2;8:643997. doi: 10.3389/fmolb.2021.643997. eCollection 2021.
LncRNAs are a new modulator in the development of intervertebral disc degeneration. However, the functional role and mechanism of HOXC13-AS in intervertebral disc degeneration remain unclear. qRT-PCR analysis was performed to measure the relative expression levels of HOXC13-AS and miR-497-5p, and the levels of IL-1β, IL-6, and TNF-α in the medium supernatant were analyzed by ELISA. The related mechanism between HOXC13-AS and miR-497-5p was detected by luciferase assays. The results revealed that TNF-α and IL-1β induced HOXC13-AS expression in NP cells. HOXC13-AS was overexpressed in IDD specimens compared to control specimens, and higher expression of HOXC13-AS was correlated with high Pfirrmann scores. Ectopic expression of HOXC13-AS promoted MMP-3 and ADAMTS4 and inhibited aggrecan and collagen II expression in NP cells. Furthermore, overexpression of HOXC13-AS increased the expression of inflammatory cytokines, including IL-1β, IL-6, and TNF-α. Our results demonstrated that TNF-α and IL-1β induced ADAMTS5 expression and suppressed miR-497-5p expression. miR-497-5p was downregulated in IDD specimens compared to control specimens, and the lower expression of miR-497-5p was correlated with high Pfirrmann scores. The miR-497-5p level was negatively proportional to HOXC13-AS expression in IDD specimens. Luciferase analysis data indicated that ADAMTS5 was a direct target gene of miR-497-5p. HOXC13-AS induced inflammatory cytokine expression and ECM degradation by modulating miR-497-5p/ADAMTS5. HOXC13-AS may be a treatment target for IDD.
长链非编码RNA是椎间盘退变发展过程中的一种新型调节因子。然而，HOXC13-AS在椎间盘退变中的功能作用和机制仍不清楚。进行qRT-PCR分析以测量HOXC13-AS和miR-497-5p的相对表达水平，并通过ELISA分析培养基上清液中IL-1β、IL-6和TNF-α的水平。通过荧光素酶报告基因检测法检测HOXC13-AS与miR-497-5p之间的相关机制。结果显示，TNF-α和IL-1β可诱导髓核细胞中HOXC13-AS的表达。与对照标本相比，HOXC13-AS在椎间盘退变标本中过表达，且HOXC13-AS的高表达与高Pfirrmann评分相关。HOXC13-AS的异位表达促进了髓核细胞中MMP-3和ADAMTS4的表达，并抑制了聚集蛋白聚糖和胶原蛋白II的表达。此外，HOXC13-AS的过表达增加了包括IL-1β、IL-6和TNF-α在内的炎性细胞因子的表达。我们的结果表明，TNF-α和IL-1β可诱导ADAMTS5的表达并抑制miR-497-5p的表达。与对照标本相比，miR-497-5p在椎间盘退变标本中表达下调，且miR-497-5p的低表达与高Pfirrmann评分相关。在椎间盘退变标本中，miR-497-5p水平与HOXC13-AS表达呈负相关。荧光素酶分析数据表明，ADAMTS5是miR-497-5p的直接靶基因。HOXC13-AS通过调节miR-497-5p/ADAMTS5诱导炎性细胞因子表达和细胞外基质降解。HOXC13-AS可能是椎间盘退变的一个治疗靶点。
