Knockdown of long non‑coding RNA KCNQ1OT1 suppresses the progression of osteoarthritis by mediating the miR‑211‑5p/TCF4 axis <em>in vitro</em>

Aili,Dilihumaer; Wu,Tong; Gu,Yuan; Chen,Ziyuan; Wang,Wanchun

doi:10.3892/etm.2021.9886

Knockdown of long non‑coding RNA KCNQ1OT1 suppresses the progression of osteoarthritis by mediating the miR‑211‑5p/TCF4 axis in vitro

Authors:
- Dilihumaer Aili
- Tong Wu
- Yuan Gu
- Ziyuan Chen
- Wanchun Wang
View Affiliations

Affiliations: Department of Orthopaedics, The Second Xiangya Hospital of Central South University, Changsha, Hunan 410011, P.R. China
Published online on: March 1, 2021 https://doi.org/10.3892/etm.2021.9886
Article Number: 455

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

Numerous studies have reported the critical roles of long non‑coding RNAs (lncRNAs) in the regulation of osteoarthritis (OA) development. The present study aimed to assess the function and regulatory mechanism of a lncRNA, KCNQ1 opposite strand/antisense transcript 1 (KCNQ1OT1), in OA in vitro. C28/I2 cells were treated with lipopolysaccharide (LPS) to generate an in vitro OA model. The relative expression levels of KCNQ1OT1, microRNA (miR)‑211‑5p and transcription factor 4 (TCF4) were determined via reverse transcription‑quantitative polymerase chain reaction. The associations between KCNQ1OT1, miR‑211‑5p and TCF4 were confirmed using a dual‑luciferase reporter assay. Furthermore, cell viability was assessed using the MTT assay. Inflammatory cytokine levels were measured using ELISA. The protein expression levels of matrix metalloproteinase‑3/13, collagen II/X and TCF4 were detected by western blotting. KCNQ1OT1 and TCF4 were highly expressed in the cartilage tissues of patients with OA and C28/I2 cells treated with LPS (OA cells), whereas miR‑211‑5p was downregulated concomitantly in OA tissues and cells. Knockdown of KCNQ1OT1 stimulated cell viability, and suppressed the inflammation and degradation of the extracellular matrix (ECM) in OA cells. In addition, overexpression of miR‑211‑5p stimulated cell viability, and inhibited inflammation and degradation of the ECM in OA cells. Notably, miR‑211‑5p was revealed to be the target of, and was negatively regulated by, KCNQ1OT1. TCF4 was targeted and negatively modulated by miR‑211‑5p. Transfection of cells with the miR‑211‑5p inhibitor or pcDNA‑TCF4 reversed the suppressive effects of short hairpin RNA (sh)‑KCNQ1OT1 on inflammation and ECM degradation, as well as the promotive effect of sh‑KCNQ1OT1 on viability in OA in vitro. Therefore, KCNQ1OT1 may regulate the miR‑211‑5p/TCF4 axis to ameliorate OA in vitro.

View Figures

View References

1	Ashford S and Williard J: Osteoarthritis: A review. Nurse Pract. 39:1–8. 2014.PubMed/NCBI View Article : Google Scholar
2	Madry H, Luyten FP and Facchini A: Biological aspects of early osteoarthritis. Knee Surg Sports Traumatol Arthrosc. 20:407–422. 2012.PubMed/NCBI View Article : Google Scholar
3	Xia B, Di C, Zhang J, Hu S, Jin H and Tong P: Osteoarthritis pathogenesis: A review of molecular mechanisms. Calcif Tissue Int. 95:495–505. 2014.PubMed/NCBI View Article : Google Scholar
4	Woolf AD and Pfleger B: Burden of major musculoskeletal conditions. Bull World Health Organ. 81:646–656. 2003.PubMed/NCBI
5	Migliore A, Paoletta M, Moretti A, Liguori S and Iolascon G: The perspectives of intra-articular therapy in the management of osteoarthritis. Expert Opin Drug Deliv. 17:1213–1226. 2020.PubMed/NCBI View Article : Google Scholar
6	Taruc-Uy RL and Lynch SA: Diagnosis and treatment of osteoarthritis. Prim Care. 40:821–836, vii. 2013.PubMed/NCBI View Article : Google Scholar
7	Fritah S, Niclou SP and Azuaje F: Databases for lncRNAs: A comparative evaluation of emerging tools. RNA. 20:1655–1665. 2014.PubMed/NCBI View Article : Google Scholar
8	Chen WK, Yu XH, Yang W, Wang C, He WS, Yan YG, Zhang J and Wang WJ: lncRNAs: Novel players in intervertebral disc degeneration and osteoarthritis. Cell Prolif. 50(e12313)2017.PubMed/NCBI View Article : Google Scholar
9	Liu Y, Liu K, Tang C, Shi Z, Jing K and Zheng J: Long non-coding RNA XIST contributes to osteoarthritis progression via miR-149-5p/DNMT3A axis. Biomed Pharmacother. 128(110349)2020.PubMed/NCBI View Article : Google Scholar
10	Chen Y, Zhang L, Li E, Zhang G, Hou Y, Yuan W, Qu W and Ding L: Long-chain non-coding RNA HOTAIR promotes the progression of osteoarthritis via sponging miR-20b/PTEN axis. Life Sci. 253(117685)2020.PubMed/NCBI View Article : Google Scholar
11	Zhang X, Huang CR, Pan S, Pang Y, Chen YS, Zha GC, Guo KJ and Zheng X: Long non-coding RNA SNHG15 is a competing endogenous RNA of miR-141-3p that prevents osteoarthritis progression by upregulating BCL2L13 expression. Int Immunopharmacol. 83(106425)2020.PubMed/NCBI View Article : Google Scholar
12	Liu C, Gao J, Su G, Xiang Y and Wan L: MicroRNA-1202 plays a vital role in osteoarthritis via KCNQ1OT1 has-miR-1202-ETS1 regulatory pathway. J Orthop Surg Res. 15(130)2020.PubMed/NCBI View Article : Google Scholar
13	Swingler TE, Niu L, Smith P, Paddy P, Le L, Barter MJ, Young DA and Clark IM: The function of microRNAs in cartilage and osteoarthritis. Clin Exp Rheumatol. 37 (Suppl 120):S40–S47. 2019.PubMed/NCBI
14	Iliopoulos D, Malizos KN, Oikonomou P and Tsezou A: Integrative microRNA and proteomic approaches identify novel osteoarthritis genes and their collaborative metabolic and inflammatory networks. PLoS One. 3(e3740)2008.PubMed/NCBI View Article : Google Scholar
15	Luo C, Liang JS, Gong J, Zhang HL, Feng ZJ, Yang HT, Zhang HB and Kong QH: The function of microRNA-34a in osteoarthritis. Bratisl Lek Listy. 120:386–391. 2019.PubMed/NCBI View Article : Google Scholar
16	Zhong G, Long H, Ma S, Shunhan Y, Li J and Yao J: Corrigendum to ‘miRNA-335-5p relieves chondrocyte inflammation by activating autophagy in osteoarthritis’ [Life Sci 226 (2019) 164-172]. Life Sci. 240(117135)2020.PubMed/NCBI View Article : Google Scholar
17	Huang J, Zhao L, Fan Y, Liao L, Ma PX, Xiao G and Chen D: The microRNAs miR-204 and miR-211 maintain joint homeostasis and protect against osteoarthritis progression. Nat Commun. 10(2876)2019.PubMed/NCBI View Article : Google Scholar
18	Prasadam I, Batra J, Perry S, Gu W, Crawford R and Xiao Y: Systematic identification, characterization and target gene analysis of microRNAs involved in osteoarthritis subchondral bone pathogenesis. Calcif Tissue Int. 99:43–55. 2016.PubMed/NCBI View Article : Google Scholar
19	Liu H and Luo J: miR-211-5p contributes to chondrocyte differentiation by suppressing Fibulin-4 expression to play a role in osteoarthritis. J Biochem. 166:495–502. 2019.PubMed/NCBI View Article : Google Scholar
20	Altman R, Asch E, Bloch D, Bole G, Borenstein D, Brandt K, Christy W, Cooke TD, Greenwald R, Hochberg M, et al: Development of criteria for the classification and reporting of osteoarthritis. Classification of osteoarthritis of the knee. Diagnostic and Therapeutic Criteria Committee of the American Rheumatism Association. Arthritis Rheum. 29:1039–1049. 1986.PubMed/NCBI View Article : Google Scholar
21	Hu Y, Li S and Zou Y: Knockdown of lncRNA H19 relieves LPS-induced damage by modulating miR-130a in osteoarthritis. Yonsei Med J. 60:381–388. 2019.PubMed/NCBI View Article : Google Scholar
22	Luo X, Wang J, Wei X, Wang S and Wang A: Knockdown of lncRNA MFI2-AS1 inhibits lipopolysaccharide-induced osteoarthritis progression by miR-130a-3p/TCF4. Life Sci. 240(117019)2020.PubMed/NCBI View Article : Google Scholar
23	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.PubMed/NCBI View Article : Google Scholar
24	Endo Y, Nwawka OK, Smith S and Burket JC: Tarsometatarsal joint communication during fluoroscopy-guided therapeutic joint injections and relationship with patient age and degree of osteoarthritis. Skeletal Radiol. 47:271–277. 2018.PubMed/NCBI View Article : Google Scholar
25	Li X, Huang TL, Zhang GD, Jiang JT and Guo PY: lncRNA ANRIL impacts the progress of osteoarthritis via regulating proliferation and apoptosis of osteoarthritis synoviocytes. Eur Rev Med Pharmacol Sci. 23:9729–9737. 2019.PubMed/NCBI View Article : Google Scholar
26	Zhang H, Li J, Shao W and Shen N: lncRNA CTBP1-AS2 is upregulated in osteoarthritis and increases the methylation of miR-130a gene to inhibit chondrocyte proliferation. Clin Rheumatol. 39:3473–3478. 2020.PubMed/NCBI View Article : Google Scholar
27	Zhu JK, He TD, Wei ZX and Wang YM: lncRNA FAS-AS1 promotes the degradation of extracellular matrix of cartilage in osteoarthritis. Eur Rev Med Pharmacol Sci. 22:2966–2972. 2018.PubMed/NCBI View Article : Google Scholar
28	Chen B, Ma J, Li C and Wang Y: Long noncoding RNA KCNQ1OT1 promotes proliferation and epithelialmesenchymal transition by regulation of SMAD4 expression in lens epithelial cells. Mol Med Rep. 18:16–24. 2018.PubMed/NCBI View Article : Google Scholar
29	Li X, Dai Y, Yan S, Shi Y, Han B, Li J, Cha L and Mu J: Down-regulation of lncRNA KCNQ1OT1 protects against myocardial ischemia/reperfusion injury following acute myocardial infarction. Biochem Biophys Res Commun. 491:1026–1033. 2017.PubMed/NCBI View Article : Google Scholar
30	Ye B, Wu ZH, Tsui TY, Zhang BF, Su X, Qiu YH and Zheng XT: lncRNA KCNQ1OT1 suppresses the inflammation and proliferation of vascular smooth muscle cells through IkappaBa in intimal hyperplasia. Mol Ther Nucleic Acids. 20:62–72. 2020.PubMed/NCBI View Article : Google Scholar
31	Liu Y, Lin L, Zou R, Wen C, Wang Z and Lin F: MSC-derived exosomes promote proliferation and inhibit apoptosis of chondrocytes via lncRNA-KLF3-AS1/miR-206/GIT1 axis in osteoarthritis. Cell Cycle. 17:2411–2422. 2018.PubMed/NCBI View Article : Google Scholar
32	Cao L, Wang Y, Wang Q and Huang J: lncRNA FOXD2-AS1 regulates chondrocyte proliferation in osteoarthritis by acting as a sponge of miR-206 to modulate CCND1 expression. Biomed Pharmacother. 106:1220–1226. 2018.PubMed/NCBI View Article : Google Scholar
33	Xu K, Meng Z, Xian XM, Deng MH, Meng QG, Fang W, Zhang D and Long X: lncRNA PVT1 induces chondrocyte apoptosis through upregulation of TNF-alpha in synoviocytes by sponging miR-211-3p. Mol Cell Probes: 101560, 2020.
34	Li L, Lv G, Wang B and Kuang L: The role of lncRNA XIST/miR-211 axis in modulating the proliferation and apoptosis of osteoarthritis chondrocytes through CXCR4 and MAPK signaling. Biochem Biophys Res Commun. 503:2555–2562. 2018.PubMed/NCBI View Article : Google Scholar
35	Wu L, Guo H, Sun K, Zhao X, Ma T and Jin Q: Sclerostin expression in the subchondral bone of patients with knee osteoarthritis. Int J Mol Med. 38:1395–1402. 2016.PubMed/NCBI View Article : Google Scholar
36	Ma B, Zhong L, van Blitterswijk CA, Post JN and Karperien M: T cell factor 4 is a pro-catabolic and apoptotic factor in human articular chondrocytes by potentiating nuclear factor kappaB signaling. J Biol Chem. 288:17552–17558. 2013.PubMed/NCBI View Article : Google Scholar
37	Xue H, Tu Y, Ma T, Wen T, Yang T, Xue L, Cai M, Wang F and Guan M: miR-93-5p attenuates IL-1β-induced chondrocyte apoptosis and cartilage degradation in osteoarthritis partially by targeting TCF4. Bone. 123:129–136. 2019.PubMed/NCBI View Article : Google Scholar
38	Wang J, Fang L, Ye L, Ma S, Huang H, Lan X and Ma J: miR-137 targets the inhibition of TCF4 to reverse the progression of osteoarthritis through the AMPK/NF-κB signaling pathway. Biosci Rep. 40(BSR20200466)2020.PubMed/NCBI View Article : Google Scholar