Department of Orthopedics, Second Hospital of Jilin University, Changchun, 132000, People's Republic of China.
Department of Orthopedics, China-Japan Union Hospital of Jilin University, Changchun, 130033, Jilin Province, People's Republic of China.
Calcif Tissue Int. 2021 Feb;108(2):252-264. doi: 10.1007/s00223-020-00760-w. Epub 2020 Oct 28.
MicroRNAs (miRNAs/miRs) are important biomarkers for the progression of intervertebral disc degeneration (IDD). We investigated the role of miR-30d in IDD progression through its interactions with forkhead box O3 (FOXO3) and C-X-C motif ligand 10 (CXCL10). We first measured the expression of miR-30d, FOXO3, and CXCL10 in NP cells cultured from IDD patients. RNA-immunoprecipitation (RIP), chromatin immunoprecipitation (ChIP) and dual-luciferase reporter assays were then employed to test the relationship among miR-30d, FOXO3, and CXCL10. Besides, gain- and loss-of function approaches were performed to assess the functional roles of miR-30d and FOXO3 in IDD in vitro and in vivo. We found high expression of miR-30d and CXCL10 and low expression of FOXO3 in IDD. We showed that miR-30d specifically targeted FOXO3, and that down-regulation of miR-30d promoted proliferation and inhibited apoptosis of NP cells in IDD by increasing the expression of FOXO3. Besides, FOXO3 inhibited apoptosis of NP cells by downregulation of CXCL10 expression. Moreover, inhibition of miR-30d promoted proliferation and inhibited apoptosis of NP cells in IDD by decreasing CXCL10. Furthermore, findings in the mouse IDD model confirmed the inhibitory role of decreased miR-30d in IDD progression. Thus, we show that downregulation of miR-30d could promote the proliferation of NP cells by increasing FOXO3 and decreasing CXCL10 expression, which may provide a novel therapeutic target for IDD.
微小 RNA（miRNAs/miRs）是椎间盘退行性变（IDD）进展的重要生物标志物。我们通过研究 miR-30d 与叉头框 O3（FOXO3）和 C-X-C 基序配体 10（CXCL10）的相互作用，探讨了其在 IDD 进展中的作用。我们首先测量了来自 IDD 患者的 NP 细胞中 miR-30d、FOXO3 和 CXCL10 的表达。然后采用 RNA 免疫沉淀（RIP）、染色质免疫沉淀（ChIP）和双荧光素酶报告基因检测来检验 miR-30d、FOXO3 和 CXCL10 之间的关系。此外，还采用了增益和缺失功能方法来评估 miR-30d 和 FOXO3 在体外和体内 IDD 中的功能作用。我们发现 IDD 中 miR-30d 和 CXCL10 的表达升高，而 FOXO3 的表达降低。我们表明，miR-30d 特异性靶向 FOXO3，下调 miR-30d 通过增加 FOXO3 的表达促进 IDD 中 NP 细胞的增殖并抑制其凋亡。此外，FOXO3 通过下调 CXCL10 的表达抑制 NP 细胞的凋亡。此外，miR-30d 的抑制作用通过降低 CXCL10 的表达促进 IDD 中 NP 细胞的增殖和抑制其凋亡。此外，在小鼠 IDD 模型中的研究结果证实了 miR-30d 的下调在 IDD 进展中的抑制作用。因此，我们表明下调 miR-30d 通过增加 FOXO3 和降低 CXCL10 的表达促进 NP 细胞的增殖，这可能为 IDD 提供了一个新的治疗靶点。
