MicroRNA‑200c‑3p suppresses intervertebral disc degeneration by targeting RAP2C/ERK signaling

Cao,Jianping; Jiang,Meng; Ren,Huafeng; Xu,Kai

doi:10.3892/mmr.2021.12505

MicroRNA‑200c‑3p suppresses intervertebral disc degeneration by targeting RAP2C/ERK signaling

Authors:
- Jianping Cao
- Meng Jiang
- Huafeng Ren
- Kai Xu
View Affiliations

Affiliations: Department of Anesthesiology, Qingdao No. 6 People's Hospital, Qingdao, Shandong 266033, P.R. China, Department of Orthopedics, Qingdao No. 6 People's Hospital, Qingdao, Shandong 266033, P.R. China, Department of Functional Inspection, Qingdao Haici Medical Treatment Group, Qingdao, Shandong 266033, P.R. China
Published online on: October 20, 2021 https://doi.org/10.3892/mmr.2021.12505
Article Number: 865
Copyright: © Cao et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Intervertebral disc degeneration (IDD) is a major cause of lower back pain. The high morbidity associated with this disease diminishes the quality of life of those who are affected. MicroRNAs (miRs) play crucial roles in various diseases, including IDD. However, the mechanism via which miR‑200c‑3p plays a role in the development of IDD remains unknown. The present study aimed to investigate the effect of miR‑200c‑3p on the progression of IDD and the underlying mechanism. The expression level of miR‑200c‑3p was evaluated in intervertebral disc tissues from patients with IDD. To construct the IDD cell model, the nucleus pulposus (NP) cells were treated with lipopolysaccharide (LPS) 24 h following transfection with miR‑200c‑3p mimic or inhibitor. A luciferase activity assay was performed, while reverse transcription‑quantitative PCR and western blotting were conducted to determine the RNA and protein expression levels, respectively. The expression level of miR‑200c‑3p in the intervertebral disc tissues of patients with IDD was lower than that of normal subjects. LPS treatment reduced the expression level of miR‑200c‑3p in NP cells. Moreover, miR‑200c‑3p mimic inhibited LPS‑induced NP cell apoptosis. It was found that miR‑200c‑3p attenuated inflammatory cytokine levels and extracellular matrix (ECM) degradation in NP cells. Furthermore, miR‑200c‑3p targeted Ras‑related protein 2C (RAP2C) in NP cells. RAP2C promoted apoptosis, inflammatory cytokine levels and ECM degradation by activating ERK signaling. Knockdown of RAP2C and inhibition of ERK signaling by SCH772984 partially reversed the proinflammatory effect of the miR‑200c‑3p inhibitor on LPS‑treated NP cells. Thus, miR‑200c‑3p inhibits NP cell apoptosis, inflammatory cytokine levels and ECM degradation in IDD by targeting RAP2C/ERK signaling.

View Figures

View References

1	Wang SZ, Chang Q, Lu J and Wang C: Growth factors and platelet-rich plasma: Promising biological strategies for early intervertebral disc degeneration. Int Orthop. 39:927–934. 2015. View Article : Google Scholar : PubMed/NCBI
2	Tisherman R, Coelho P, Phillibert D, Wang D, Dong Q, Vo N, Kang J and Sowa G: NF-κB signaling pathway in controlling intervertebral disk cell response to inflammatory and mechanical stressors. Phys Ther. 96:704–711. 2016. View Article : Google Scholar : PubMed/NCBI
3	Zhang Y, Yang J, Zhou X, Wang N, Li Z, Zhou Y, Feng J, Shen D and Zhao W: Knockdown of miR-222 inhibits inflammation and the apoptosis of LPS-stimulated human intervertebral disc nucleus pulposus cells. Int J Mol Med. 44:1357–1365. 2019.PubMed/NCBI
4	Zhou Q, Huang SX, Zhang F, Li SJ, Liu C, Xi YY, Wang L, Wang X, He QQ, Sun CC and Li DJ: MicroRNAs: A novel potential biomarker for diagnosis and therapy in patients with non-small cell lung cancer. Cell Prolif. 50:e123942017. View Article : Google Scholar : PubMed/NCBI
5	Zhang Q, Weng Y, Jiang Y, Zhao S, Zhou D and Xu N: Overexpression of miR-140-5p inhibits lipopolysaccharide-induced human intervertebral disc inflammation and degeneration by downregulating toll-like receptor 4. Oncol Rep. 40:793–802. 2018.PubMed/NCBI
6	Liu W, Xia P, Feng J, Kang L, Huang M, Wang K, Song Y, Li S, Wu X, Yang S and Yang C: MicroRNA-132 upregulation promotes matrix degradation in intervertebral disc degeneration. Exp Cell Res. 359:39–49. 2017. View Article : Google Scholar : PubMed/NCBI
7	Yu G, Jiao Y, Huang JJ, Fan MD, Hao YC, Han JZ and Qu L: Acidic preconditioning reduces lipopolysaccharide-induced acute lung injury by upregulating the expression of angiotensin-converting enzyme 2. Exp Ther Med. 21:4412021. View Article : Google Scholar : PubMed/NCBI
8	Lai Z and Cao Y: Plasma miR-200c-3p, miR-100-5p, and miR-1826 serve as potential diagnostic biomarkers for knee osteoarthritis: Randomized controlled trials. Medicine (Baltimore). 98:e181102019. View Article : Google Scholar : PubMed/NCBI
9	Chen P, Jiang P, Chen J, Yang Y and Guo X: XIST promotes apoptosis and the inflammatory response in CSE-stimulated cells via the miR-200c-3p/EGR3 axis. BMC Pulm Med. 21:021–01582. 2021. View Article : Google Scholar
10	Wang Z, Huang C, Zhang A, Lu C and Liu L: Overexpression of circRNA_100290 promotes the progression of laryngeal squamous cell carcinoma through the miR-136-5p/RAP2C axis. Biomed Pharmacother. 125:1098742020. View Article : Google Scholar : PubMed/NCBI
11	Wu J, Du W, Wang X, Wei L, Pan Y, Wu X, Zhang J and Pei D: Ras-related protein Rap2c promotes the migration and invasion of human osteosarcoma cells. Oncol Lett. 15:5352–5358. 2018.PubMed/NCBI
12	Wang Z, Huang C, Zhang A, Lu C and Liu L: Overexpression of circRNA_100290 promotes the progression of laryngeal squamous cell carcinoma through the miR-136-5p/RAP2C axis. Biomed Pharmacother. 125:109872020. View Article : Google Scholar
13	Wang X, Wang C, Xi L and Yu Z: Rap2c as a novel biomarker for predicting poor prognosis in glioma. Onco Targets Ther. 13:3073–3083. 2020. View Article : Google Scholar : PubMed/NCBI
14	Mi D, Cai C, Zhou B, Liu X, Ma P, Shen S, Lu W and Huang W: Long noncoding RNA FAF1 promotes intervertebral disc degeneration by targeting the Erk signaling pathway. Mol Med Rep. 17:3158–3163. 2018.PubMed/NCBI
15	Li L, Wei K, Ding Y, Ahati P, Xu H, Fang H and Wang H: M2a macrophage-secreted CHI3L1 promotes extracellular matrix metabolic imbalances via activation of IL-13Rα2/MAPK pathway in rat intervertebral disc degeneration. Front Immunol. 12:6663612021. View Article : Google Scholar : PubMed/NCBI
16	Ni L, Zheng Y, Gong T, Xiu C, Li K, Saijilafu, Li B, Yang H and Chen J: Proinflammatory macrophages promote degenerative phenotypes in rat nucleus pulpous cells partly through ERK and JNK signaling. J Cell Physiol. 234:5362–5371. 2019. View Article : Google Scholar : PubMed/NCBI
17	Pfirrmann CW, Metzdorf A, Zanetti M, Hodler J and Boos N: Magnetic resonance classification of lumbar intervertebral disc degeneration. Spine (Phila Pa 1976). 26:1873–1878. 1976. View Article : Google Scholar
18	Ji Y, Hong W, Liu M, Liang Y, Deng Y and Ma L: Intervertebral disc degeneration associated with vertebral marrow fat, assessed using quantitative magnetic resonance imaging. Skeletal Radiol. 49:1753–1763. 2020. View Article : Google Scholar : PubMed/NCBI
19	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
20	Yang Y, Zhong Z, Zhao Y, Ren K and Li N: LincRNA-SLC20A1 (SLC20A1) promotes extracellular matrix degradation in nucleus pulposus cells in human intervertebral disc degeneration by targeting the miR-31-5p/MMP3 axis. Int J Clin Exp Pathol. 12:3632–3643. 2019.PubMed/NCBI
21	Navone SE, Marfia G, Giannoni A, Beretta M, Guarnaccia L, Gualtierotti R, Nicoli D, Rampini P and Campanella R: Inflammatory mediators and signalling pathways controlling intervertebral disc degeneration. Histol Histopathol. 32:523–542. 2017.PubMed/NCBI
22	Han YC, Ma B, Guo S, Yang M, Li LJ, Wang SJ and Tan J: Leptin regulates disc cartilage endplate degeneration and ossification through activation of the MAPK-ERK signalling pathway in vivo and in vitro. J Cell Mol Med. 22:2098–2109. 2018. View Article : Google Scholar : PubMed/NCBI
23	Roskoski R Jr: ERK1/2 MAP kinases: Structure, function, and regulation. Pharmacol Res. 66:105–143. 2012. View Article : Google Scholar : PubMed/NCBI
24	Roskoski R Jr: Targeting ERK1/2 protein-serine/threonine kinases in human cancers. Pharmacol Res. 142:151–168. 2019. View Article : Google Scholar : PubMed/NCBI
25	Sampara P, Banala RR, Vemuri SK, Av GR and Gpv S: Understanding the molecular biology of intervertebral disc degeneration and potential gene therapy strategies for regeneration: A review. Gene Ther. 25:67–82. 2018. View Article : Google Scholar : PubMed/NCBI
26	Shen L, Xiao Y, Wu Q, Liu L, Zhang C and Pan X: TLR4/NF-κB axis signaling pathway-dependent up-regulation of miR-625-5p contributes to human intervertebral disc degeneration by targeting COL1A1. Am J Transl Res. 11:1374–1388. 2019.PubMed/NCBI
27	Tan H, Zhao L, Song R, Liu Y and Wang L: microRNA-665 promotes the proliferation and matrix degradation of nucleus pulposus through targeting GDF5 in intervertebral disc degeneration. J Cell Biochem. 119:7218–7225. 2018. View Article : Google Scholar : PubMed/NCBI
28	Chen Z, Liu M, Zhang W, Deng M, Zhou Y and Li Y: miR-24-3p induces human intervertebral disc degeneration by targeting insulin-like growth factor binding protein 5 and the ERK signaling pathway. Life Sci. 243:1172882020. View Article : Google Scholar : PubMed/NCBI
29	Dong W, Liu J, Lv Y, Wang F, Liu T, Sun S, Liao B, Shu Z and Qian J: miR-640 aggravates intervertebral disc degeneration via NF-κB and WNT signalling pathway. Cell Prolif. 52:e126642019. View Article : Google Scholar : PubMed/NCBI
30	Lagarrigue F, Kim C and Ginsberg MH: The Rap1-RIAM-talin axis of integrin activation and blood cell function. Blood. 128:479–487. 2016. View Article : Google Scholar : PubMed/NCBI
31	Paganini S, Guidetti GF, Catricalà S, Trionfini P, Panelli S, Balduini C and Torti M: Identification and biochemical characterization of Rap2C, a new member of the Rap family of small GTP-binding proteins. Biochimie. 88:285–295. 2006. View Article : Google Scholar : PubMed/NCBI
32	Liao X, Zhan W, Tian B, Luo Y, Gu F and Li R: Circular RNA ZNF609 promoted hepatocellular carcinoma progression by upregulating PAP2C expression via sponging miR-342-3p. Onco Targets Ther. 13:7773–7783. 2020. View Article : Google Scholar : PubMed/NCBI
33	Shen Z, Zhou R, Liu C, Wang Y, Zhan W, Shao Z, Liu J, Zhang F, Xu L, Zhou X, et al: MicroRNA-105 is involved in TNF-α-related tumor microenvironment enhanced colorectal cancer progression. Cell Death Dis. 8:32132017. View Article : Google Scholar : PubMed/NCBI
34	Qiao Y, Peng C, Li J, Wu D and Wang X: Spinal cord ischemia-reperfusion causes damage of neurocyte by inhibiting RAP2C. Neurol Res. 39:877–884. 2017. View Article : Google Scholar : PubMed/NCBI
35	Zhu X, Qiu J, Zhang T, Yang Y, Guo S, Li T, Jiang K, Zahoor A, Deng G and Qiu C: MicroRNA-188-5p promotes apoptosis and inhibits cell proliferation of breast cancer cells via the MAPK signaling pathway by targeting Rap2c. J Cell Physiol. 235:2389–2402. 2020. View Article : Google Scholar : PubMed/NCBI
36	Chen Y, Zheng Z, Wang J, Tang C, Khor S, Chen J, Chen X, Zhang Z, Tang Q, Wang C, et al: Berberine suppresses apoptosis and extracellular matrix (ECM) degradation in nucleus pulposus cells and ameliorates disc degeneration in a rodent model. Int J Biol Sci. 14:682–692. 2018. View Article : Google Scholar : PubMed/NCBI
37	Ruan Z, Ma H, Li J, Liu H, Jia H and Li F: The long non-coding RNA NEAT1 contributes to extracellular matrix degradation in degenerative human nucleus pulposus cells. Exp Biol Med (Maywood). 243:595–600. 2018. View Article : Google Scholar : PubMed/NCBI
38	Zhang J, Li Z, Chen F, Liu H, Wang H, Li X, Liu X, Wang J and Zheng Z: TGF-β1 suppresses CCL3/4 expression through the ERK signaling pathway and inhibits intervertebral disc degeneration and inflammation-related pain in a rat model. Exp Mol Med. 49:e3792017. View Article : Google Scholar : PubMed/NCBI
39	Li S, Hua W, Wang K, Gao Y, Chen S, Liu W, Song Y, Wu X, Tu J, Kang L, et al: Autophagy attenuates compression-induced apoptosis of human nucleus pulposus cells via MEK/ERK/NRF1/Atg7 signaling pathways during intervertebral disc degeneration. Exp Cell Res. 370:87–97. 2018. View Article : Google Scholar : PubMed/NCBI