Graduate School of Inner Mongolia Medical University, Hohhot City, 010059, Inner Mongolia, China.
Department of Thoracolumbar Spine Surgery, The Second Affiliated Hospital of Inner Mongolia Medical University, No. 1 Yingfang Road, Hohhot City, 010059, Inner Mongolia, China.
J Orthop Surg Res. 2021 Jan 14;16(1):55. doi: 10.1186/s13018-020-02194-4.
Intervertebral disk degeneration (IDD) is a degenerative disease characterized by cytoplasm loss and extracellular matrix degradation. Numerous evidence reported that miRNAs participated in IDD development. Nevertheless, the function of miR-142-3p in IDD development remains unknown. This study mainly explored the potential role and function of miR-142-3p in IDD development.
One percent fetal bovine serum was used to induce the degeneration of ATDC5 cells, and miR-142-3p level was examined by qRT-PCR. Then, miR-142-3p mimic/inhibitor and its corresponding negative control were transfected into ATDC5 normal and degenerative cells. Viability, migration, invasion, apoptosis, cycle, Bax, Bcl-2, P62, and Beclin1 expression levels were assessed using CCK8, wound healing assay, annexin V-FITC/PI staining, western blot, and qRT-PCR, respectively.
The results revealed that the expression levels of MMP13, ADAMTS5, MMP3, and Col-X were increased as well as the expression levels of SOX-9 and Col-II were reduced in ATDC5 degenerative cells, indicating the degeneration model was constructed. We observed that miR-142-3p was decreased in ATDC5 degenerative cells and its suppression could promote ATDC5 cell degeneration. However, miR-142-3p overexpression could reverse the cell viability inhibition, as well as apoptosis and autophagy enhancement in ATDC5 degenerative cells.
Our results proved that miR-142-3p may play an important role in disk degeneration. Further animal study is needed to illustrate the role of the miR-142-3p in IDD development.
椎间盘退变（IDD）是一种以细胞浆丢失和细胞外基质降解为特征的退行性疾病。大量证据表明，miRNAs 参与了 IDD 的发生发展。然而，miR-142-3p 在 IDD 发生发展中的作用尚不清楚。本研究主要探讨了 miR-142-3p 在 IDD 发生发展中的潜在作用和功能。
用 1%胎牛血清诱导 ATDC5 细胞退变，并用 qRT-PCR 检测 miR-142-3p 水平。然后，将 miR-142-3p 模拟物/抑制剂及其相应的阴性对照转染到 ATDC5 正常和退变细胞中。通过 CCK8 法、划痕愈合实验、 Annexin V-FITC/PI 染色、Western blot 和 qRT-PCR 分别评估细胞活力、迁移、侵袭、凋亡、细胞周期、Bax、Bcl-2、P62 和 Beclin1 的表达水平。
结果显示，ATDC5 退变细胞中 MMP13、ADAMTS5、MMP3 和 Col-X 的表达水平升高，而 SOX-9 和 Col-II 的表达水平降低，表明构建了退变模型。我们观察到 miR-142-3p 在 ATDC5 退变细胞中表达下调，其抑制可促进 ATDC5 细胞退变。然而，miR-142-3p 的过表达可以逆转 ATDC5 退变细胞活力抑制以及凋亡和自噬增强。
我们的结果证明 miR-142-3p 可能在椎间盘退变中发挥重要作用。需要进一步的动物研究来阐明 miR-142-3p 在 IDD 发生发展中的作用。
