Huang Yizhen, Gao Jun, Wang Jianle, Ye Huali, Yao Teng, Xu Yining, Chen Zizheng, Shen Shuying, Ma Jianjun
Department of Orthopaedic Surgery, Sir Run Run Shaw Hospital, Zhejiang University School of Medicine, Zhejiang, China.
Key Laboratory of Musculoskeletal System Degeneration and Regeneration Translational Research of Zhejiang Province, Zhejiang, China.
Cell Death Discov. 2021 Feb 26;7(1):39. doi: 10.1038/s41420-021-00420-4.
The molecular mechanism underlying the development of intervertebral disc disease (IVDD) is not completely understood. Circular RNAs (circRNAs) play a significant role in the occurrence and development of various diseases, and studies have shown that circPKNOX1 is involved in the compensatory response of extracellular matrix synthesis and secretion of the nucleus pulposus (NP) cells. However, the mechanism through which circRNAs regulate IVDD progression remains unclear; therefore, in this study, we explored the significance of circPKNOX1 in IVDD. The expression of circRNAs in NP cells of normal and degenerative patients was detected using microarray analysis, and the role of circPKNOX1 in IVDD was confirmed using RT-qPCR. The interaction networks of circRNAs, miRNAs, and miRNA target genes were detected using bioinformatics analysis, RNA fluorescence in situ hybridization, and immunofluorescence analysis. We found that the expression of circPKNOX1 decreased in IVDD cells. The expression of circPKNOX1 in NP cells, observed using RT-qPCR and western blotting, was consistent with that observed using array screening. Overexpression of circPKNOX1 increased the expression of collagen II, aggrecan, and SOX9 and decreased that of ADAMTS4, ADAMTS-5, MMP3, and MMP13. We further demonstrated that circPKNOX1 played the role of a sponge by competitively binding miR-370-3p to reverse the inhibition of KIAA0355 expression. Our findings indicated that circPKNOX1 affected the progression of IVDD by regulating the expression of KIAA0355 via miR-370-3p. Therefore, circPKNOX1-based therapy may serve as an effective IVDD treatment strategy.
椎间盘疾病(IVDD)发生发展的分子机制尚未完全明确。环状RNA(circRNA)在多种疾病的发生发展中发挥着重要作用,研究表明circPKNOX1参与了髓核(NP)细胞细胞外基质合成与分泌的代偿反应。然而,circRNA调节IVDD进展的机制仍不清楚;因此,在本研究中,我们探讨了circPKNOX1在IVDD中的意义。使用微阵列分析检测正常和退变患者NP细胞中circRNA的表达,并使用RT-qPCR证实circPKNOX1在IVDD中的作用。使用生物信息学分析、RNA荧光原位杂交和免疫荧光分析检测circRNA、miRNA和miRNA靶基因的相互作用网络。我们发现circPKNOX1在IVDD细胞中的表达降低。使用RT-qPCR和蛋白质印迹法观察到的NP细胞中circPKNOX1的表达与阵列筛选结果一致。circPKNOX1的过表达增加了胶原蛋白II、聚集蛋白聚糖和SOX9的表达,并降低了ADAMTS4、ADAMTS-5、MMP3和MMP13的表达。我们进一步证明,circPKNOX1通过竞争性结合miR-370-3p发挥海绵作用,以逆转对KIAA0355表达的抑制。我们的研究结果表明,circPKNOX1通过miR-370-3p调节KIAA0355的表达来影响IVDD的进展。因此,基于circPKNOX1的治疗可能是一种有效的IVDD治疗策略。