School of Graduate, Inner Mongolia Medical University, Hohhot, Inner Mongolia, China.
Surgical Department of Thoracolumbar, the Second Affiliated Hospital of Inner Mongolia Medical University, Hohhot, Inner Mongolia, China.
Cartilage. 2021 Dec;13(2_suppl):592S-603S. doi: 10.1177/19476035211012444. Epub 2021 May 6.
Cartilage endplate (CEP) degeneration plays a vital role in the pathological process of intervertebral disc degeneration. It has been previously reported that microRNAs may participate in the occurrence and development of intervertebral disc degeneration through regulating its target genes directly. The regulatory roles of miR-142-3p/HMGB1 in some orthopedic diseases have been determined successively, but there was no report about the degeneration of CEP. Therefore, we aimed to determine the regulation of miR-142-3p/HMGB1 or potential molecular mechanisms on proliferation, apoptosis, migration, and autophagy of CEP cells.
The target gene of miR-142-3p was determined by double luciferase assay. We selected ATDC5 cell lines. CCK-8 method was used to detect cell proliferation. Real-time fluorescence quantitative polymerase chain reaction was used to determine gene expression levels, and western blot analysis was used to determine protein expression levels. We chose flow cytometry to measure cell apoptosis and cell cycle.
The result of luciferase detection showed that the target gene of miR-142-3p in CEP cells was HMGB1. Knockdown of the miR-142-3p inhibited the expression level of HMGB1, the proliferation and migration of CEP cells, but it promoted apoptosis of CEP cells. In addition, the detection results of the proteins related to apoptosis or autophagy showed that knockdown of miR-142-3p promoted apoptosis and autophagy.
The negative regulation of miR-142-3p/HMGB1 can affect the proliferation, apoptosis, migration, and autophagy of CEP cells. Our results provide a new idea for the targeted treatment of CEP degeneration by inhibiting the expression of HMGB1.
软骨终板（CEP）退变在椎间盘退变的病理过程中起着至关重要的作用。先前有研究报道，microRNAs 可能通过直接调控其靶基因参与椎间盘退变的发生发展。miR-142-3p/HMGB1 在一些骨科疾病中的调控作用已相继被确定，但CEP 退变方面尚无相关报道。因此，我们旨在确定 miR-142-3p/HMGB1 或其对 CEP 细胞增殖、凋亡、迁移和自噬的潜在分子机制的调节作用。
采用双荧光素酶报告基因实验确定 miR-142-3p 的靶基因。我们选择 ATDC5 细胞系。CCK-8 法检测细胞增殖。实时荧光定量聚合酶链反应检测基因表达水平，Western blot 分析检测蛋白表达水平。采用流式细胞术检测细胞凋亡和细胞周期。
荧光素酶检测结果显示，CEP 细胞中 miR-142-3p 的靶基因是 HMGB1。miR-142-3p 的下调抑制了 HMGB1 的表达水平，CEP 细胞的增殖和迁移，但促进了 CEP 细胞的凋亡。此外，与凋亡或自噬相关的蛋白检测结果显示，miR-142-3p 的下调促进了凋亡和自噬。
miR-142-3p/HMGB1 的负调控可影响 CEP 细胞的增殖、凋亡、迁移和自噬。我们的研究结果为通过抑制 HMGB1 的表达靶向治疗 CEP 退变提供了新的思路。
