miR-142-3p and HMGB1 Are Negatively Regulated in Proliferation, Apoptosis, Migration, and Autophagy of Cartilage Endplate Cells
miR-142-3p and HMGB1 Are Negatively Regulated in Proliferation, Apoptosis, Migration, and Autophagy of Cartilage Endplate Cells
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DOI：
10.1177/19476035211012444
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发表时间：
2021-03
期刊：
影响因子：
2.8
通讯作者：
Bo Wang;Demin Ji;Wenhua Xing;Feng Li;Zhi Huang;Wenkai Zheng;Jianmin Xue;Yong Zhu;Xuejun Yang
中科院分区：
文献类型：
--
作者：
Bo Wang;Demin Ji;Wenhua Xing;Feng Li;Zhi Huang;Wenkai Zheng;Jianmin Xue;Yong Zhu;Xuejun Yang
背景软骨终板（CEP）退变在椎间盘退变的病理过程中起重要作用。此前已有报道，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节作用已被陆续确定，但在CEP退行性变中尚未见报道。因此，我们旨在确定miR-142-3p/HMGB1对CEP细胞增殖、凋亡、迁移和自噬的调控或潜在的分子机制。方法采用双荧光素酶法检测miR-142-3p的靶基因。我们选择了ATDC5细胞系。CCK-8法检测细胞增殖。采用实时荧光定量聚合酶链反应检测基因表达水平，western blot检测蛋白表达水平。我们采用流式细胞术检测细胞凋亡和细胞周期。结果荧光素酶检测结果显示，miR-142-3p在CEP细胞中的靶基因为HMGB1。miR-142-3p敲低可抑制HMGB1的表达水平，抑制CEP细胞的增殖和迁移，但可促进CEP细胞凋亡。此外，凋亡或自噬相关蛋白的检测结果显示，miR-142-3p敲低可促进细胞凋亡和自噬。结论miR-142-3p/HMGB1的负调控可影响CEP细胞的增殖、凋亡、迁移和自噬。我们的研究结果为抑制HMGB1的表达靶向治疗CEP变性提供了新的思路。
Background 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. Methods 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. Results 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. Conclusion 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.